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EUROPEAN COMMISSION Directorate-General for Research Directorate F — Health Unit F.3 — Infectious Diseases Contact: Cornelius SCHMALTZ European Commission Office [CDMA 2/137] B-1049 Brussels

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Printed on white chlorine-free paper Commissioners’ Preface

The emergence and spread of the highly pathogenic H5N1 avian influenza and its unusually high toll in sporadic cases of human has drawn the attention of the public to the health threat of a new influenza pandemic. While we know today that the ‘Spanish Flu’ pandemic of 1918 to 1919 was responsible for far more (up to 50 million) than the First World War, it is easily overlooked that even ‘regular’ seasonal influenza epidemics are responsible for an estimated 250 000 to 300 000 deaths each year worldwide. In addition to human illness and , the disease has an enormous direct and indirect economic impact on afflicted countries. The cost of a moderate influenza epidemic in France alone has recently been estimated at EUR 16.6 billion.

Similarly, avian influenza represents one of the major concerns to animal health. The disease is listed by the World Organisation for Animal Health (OIE) and is subject to EU legislation. Its incidence has greatly increased in the past 6 years compared to the previous 40 years, in particular due to the H5N1 strain. Outbreaks of highly pathogenic avian influenza cause huge economic losses to the poultry industry and directly affect food security and the livelihood of rural areas in developing countries. It is estimated that hundreds of millions of animals have died or have been culled as a consequence of the H5N1 outbreaks around the world.

Science has already delivered a number of effective tools for protection measures against influenza (such as human and animal as well as antiviral ), but very important questions remain unanswered, and research is of prime importance in the fight against influenza. We need more potent, longer-acting and more broadly protective vaccines for humans (including vaccines against a potential pandemic strain). We need to develop better veterinary vaccines that allow vaccinated and infected birds to be distinguished — crucial for effective surveillance and large scale administration. We need improved rapid diagnostic methods in birds and humans and new molecular targets for better antiviral drugs, as well as research into the effectiveness of containment and mitigation strategies in the case of a pandemic.

We would like to take this opportunity to emphasise that the European Commission’s support for influenza research dates back to long before the current media attention to the threat of an H5N1 pandemic. This catalogue gives you an overview of the breadth and the depth of this effort. Since 2001, the Commission has supported specific influenza research projects with over EUR 65 million. In addition, it is currently supporting, with more than EUR 40 million, a number of larger projects on viral or infectious diseases in general, in which influenza plays an important role. This level of commitment arguably makes the Commission the most important funding organisation for influenza research in Europe. As you will discover in this publication, several of these projects have already resulted in major scientific breakthroughs and/or have already led to improvements in public health policy.

This publication cuts across a variety of different activities in the Fifth and Sixth Framework Programmes for Research and also includes projects funded through the Programme of Community Action in the field of Public Health, illustrating the interdisciplinary nature of influenza research. It is our firm conviction that the fight against influenza — like many other challenges — can only be won if research, policy support and the implementation of research results work closely together for the benefit of public health. Many of the projects in this publication are living proof of this inextricable link.

It almost goes without saying that uniting Europe’s considerable strength in influenza research and fostering the collaboration of teams with complementary expertise is as important as joining forces in the public and veterinary health measures against a virus that — more than ever in an era of globalisation that includes human travel as well as the poultry trade — does not stop at any border. We are glad to say that we will continue to support all these efforts in the Seventh Research Framework Programme (FP7), which was launched earlier this year and which will run until 2013, and in the context of the new Public Health Programme covering the period 2008 to 2013. For the first time a specific activity within the ‘Health’ theme of FP7’s Cooperation Programme will be dedicated to ‘Emerging Infectious Epidemics’. We look forward to a future edition of this publication with the results of the first FP7-funded influenza research projects!

Janez Potočnik Markos Kyprianou European Commissioner European Commissioner in charge of Science and Research in charge of Health  INtroduction

Pandemic influenza has become a paradigm of the potential health threat that emerging infectious diseases pose to the world. Today, we could not be further from the optimism of the 1960s and 1970s, when statements like ‘the war against infectious diseases has been won’ abounded. The ‘Spanish Flu’ pandemic of 1918 to 1919 remains one of the most deadly infectious diseases of all time, and the recent spread of the highly pathogenic avian influenza virus H5N1 is just the most recent example of the continued threat of emerging zoonoses. Against this background the European Commission’s Seventh Research Framework Programme (FP7, 2007-2013) has established for the first time a new dedicated activity — ‘Emerging Infectious Epidemics’ — and the Council Decision for FP7’s Specific Programme 1. Council Decision 971/2006/EC, OJ L 400, 1 30.12.2006, p. 124 ‘Cooperation’ explicitly mentions highly pathogenic influenza in the description of this activity.

The new mandate to build a coherent strategy in this research area is also an opportunity to take stock of past and ongoing EU-funded activities in influenza research. As with previous disease outbreaks, such as BSE (bovine spongiform encephalopathy) and SARS (Sudden Acute Respiratory Syndrome), the Commission has been able to respond rapidly to the spread of bird flu and human sporadic cases of H5N1 with a EUR 28 million dedicated call for proposals, launched in late 2005. The present publication also demonstrates the longer-term track record of Framework Programme funding in this field.

The aim was to include in this catalogue all FP5- (1999-2002) and FP6- (2002-2006) funded projects that are either exclusively dedicated to research on any aspect of influenza (the majority of projects) or address a broader range of diseases, but with a significant part devoted to influenza. A table at the end of this catalogue differentiates these two groups of projects. Also included are four influenza-specific projects from outside the Research Framework Programmes. These are from the Directorate General for Health and Consumer Protection’s Programme of Community Action in the Field of Public Health. This valuable addition underlines the coherence of the Commission’s policy in this area and the close cooperation between the responsible services. The substantial contribution of a number of other public health projects that take a broader approach to infectious diseases (the training of epidemiologists, horizontal issues in vaccination policy or the improvement of communicable diseases control and communication) was also fully recognised, but it was decided, however, not to include these projects in the present publication in order to keep the focus on influenza research projects.

Given the close link between animal and human influenza, it was an obvious decision to include in this compilation projects on animal, as well as human influenza. Recent calls and future plans in the area of influenza as well as other emerging zoonoses are being coordinated in close collaboration between the units responsible for animal and human infectious diseases within the Research Directorate General.

The Framework Programmes’ emphasis on interdisciplinary research seemed to justify an attempt to divide the projects into thematic chapters that cross the border between animal and human influenza. The hope is that this structure might serve as an incentive for interesting comparisons, detecting and questioning parallel or divergent approaches. Even closer collaboration between the animal health and human health specialists will be required in the future in order to successfully combat influenza as well as other zoonoses. Together with the respective scientific communities, the Commission services will also use this compilation to identify strengths and weaknesses in our funding portfolio. Is it defensible, for example, that there is currently only one human project (from the Public Health portfolio) in the Diagnostics and Surveillance chapter? What are the lessons to be drawn from the multitude of  projects? What should be the priorities in this field for future calls? At the same time, there are of course inherent difficulties in any ex-post classification. Clearly there are projects that fall into more than one category or might in fact include additional areas not covered in the four chapters. Similarly, the ‘labels’ of those four categories, Vaccines; Diagnostics and Surveillance; Biology; and Networking, Training, Socio-Economic and Legal Issues can certainly be questioned. I apologise in advance for any possible misclassification, but hope that the advantages of this approach for the reader will outweigh its imperfections. On the last pages of the booklet some alternative classifications are provided that group the projects alphabetically, according to animal/human influenza and according to their origin in FP5, FP6 or Public Health.

CORNELIUS SCHMALTZ

Acknowledgements: Many people have contributed to this catalogue. First and foremost of course, my thanks goes to the scientific coordinators (and their staff) of the projects presented. To a very high degree they have been able to accommodate all wishes, have revised and adjusted their abstracts, have provided illustrations and been patient with my requests. Anna Lonnroth and Bernard Mulligan (Deputy Head and Acting Head of the Infectious Diseases Unit) and Tuija Jansson have provided critical support in every phase of the project. Without the enormous contribution and key advice of Isabel Minguez-Tudela, the scientific officer responsible for all animal influenza projects, this publication could never have been realised. My other colleagues from the Research Directorate General, Torbjoern Ingemansson, Annabelle Ascher, Marta Iglesias, Christian Wimmer and Beatrice Lucaroni have been very helpful in identifying and contributing influenza projects from their respective programmes. Similarly, my thanks goes to Michel Pletschette, Head of Unit, Solvejg Wallyn and Cinthia Menel Lemos from the Public Health Executive Agency, as well as to Franz Karcher and Frank van Loock from the Directorate General for Health and Consumer Protection for their crucial support regarding the Public Health projects.  Vaccines

Vaccination is considered the cornerstone of influenza control in humans. Vaccination against seasonal influenza has been in use for more than 50 years, has an excellent safety record and provides a high degree of protection. It is currently recommended in most 1. European Centre for Disease Protection and Control (ECDC): Technical Report of the Scientific Panel on countries for use in risk groups (mainly the elderly and people Vaccines and Immunisation: Infant and children with an underlying medical condition), but routine use in children1 seasonal immunisation against influenza on a routine basis during inter-pandemic period, Stockholm, and even universal immunisation programmes exist or are under January 2007 (http://www.ecdc.eu.int/documents/  consideration in some countries/regions. pdf/Flu_vacc_18_Jan.pdf) Yet, many challenges remain that are further amplified when it comes to the challenge of pandemic vaccines. Longer-acting vaccines that are more broadly cross-protective against heterologous strains and clades of influenza are required, immune responses in elderly and small children need to be optimised, and cell-culture production (which could be scaled up much faster in the case of a pandemic) is about to replace egg-based production facilities (where egg supply might be particularly vulnerable during a pandemic derived from an avian strain of influenza).

The use of vaccination in poultry is complex and only a limited number of countries have implemented a vaccination strategy to date. With currently available vaccines, avian influenza (AI) can still infect and replicate in vaccinated birds. However, they protect against clinical signs and mortality, reduce the level and duration of virus shedding and hence decrease possibilities of transmission. There is evidence that when used adequately and as part of a wider control strategy in combination with other measures, such as increased biosecurity, stamping out and appropriate surveillance, vaccination may be a powerful tool in the control and eradication of AI. A key issue for the use of vaccines is the ability to detect birds exposed to the field virus in a vaccinated population. This is known as the DIVA strategy (differentiating infected from vaccinated animals). The development of new DIVA vaccines with a wider cross-protection and species range that can be administered on a large scale and at low cost is thus an important research task.

Some of the earliest funded projects in this catalogue are vaccine projects that have delivered a number of important results: MUCADJ has developed a potent new adjuvant for mucosal (i.e. intranasal) delivery of human influenza vaccines and an H7N1 vaccine developed by the FLUPAN consortium is currently in clinical trials. A novel computer algorithm that assesses the relatedness of influenza viruses was developed by NOVAFLU, and has already been integrated into the biannual WHO (World Health Organisation) vaccine strain selection process. FLUAID works on vaccine candidate strains as well as on diagnostic tests with partners in Asia, Australia and Africa. A remarkably high number of consortia (eight) are led by industry, in particular small and medium-sized enterprises (SMEs). These are MUCADJ, CHIMERIC VACCINES, UNIVERSAL VACCINE, SARS/FLU VACCINE, FLUVACC, AIV VACC DIAGNOSIS, Intranasal H5vaccine and FluVac. The last two, as well as PANFLUVAC, are the result of the most recent (and last FP6) call for proposals. They have just started, and address the special challenges of the clinical development of a pandemic human vaccine through novel adjuvants, delivery methods and genetically engineered vaccine viruses. Similarly, AIV VACC DIAGNOSIS and NOVADUCK, funded through the same recent call, are both aimed at creating effective live influenza vaccines for poultry that allow the distinction of vaccinated and infected animals through specific markers and could be mass administered. Importantly, the Public Health project FLUSECURE is dedicated to establishing a network of public health institutes as a partner for the vaccine industry to promote the production of an effective pandemic vaccine. Efficacious vaccine formulation system for prophylactic control of influenza pandemics Key words: Pandemic H5N1 influenza, intranasal Acronym: PANFLUVAC delivery, virosomal vaccine, lipopeptide adjuvants, EC contribution: €3 334 798 Starting date: 01/01/2007 dendritic cell targeting, pre-clinical to clinical Duration: 48 months Instrument: STREP evaluation strategy

Summary: Problem: delivery and formulation to meet current and Influenza epidemics remain a burden for The challenge future influenza pandemics. The proposed H5N1 both human health and national economies, The avian H5N1 influenza virus is now spreading vaccine will be prepared with regard to the as witnessed by the recent advance of the among the wild bird population in Europe, and immediate needs, as identified above by the pathogenic avian H5N1 influenza virus. While the of humans has already reached the borders WHO. In addition, the project will generate the numbers of human deaths in Europe has remained of continental Europe. For a truly efficacious vaccine, 'mock-up' library of vaccine reagents to meet relatively low, the presence of such cases in Turkey one must consider the route of virus entry into the the permanent threat from pandemic influenza. demonstrates the danger posed by this virus. The host (the respiratory tract), and host requirements The overall aim is to construct the efficacious avian H5N1 virus has now been detected in wild for protective immune defences. That is, an ideal H5N1 vaccine, enhancing the capacity to protect birds in numerous European countries, and the vaccine should induce both local (mucosal) and the people of Europe both now and in the PANFLUVAC consortium is committed to creating systemic (serum) immunity. future, rather than just in the short term. The an efficacious vaccine against this virus, to rapidly ageing European population, which is provide strong protection in a pandemic situation. Currently, parenterally administered inactivated particularly vulnerable to influenza complications influenza vaccine is the best prophylactic control and influenza-related deaths, is a clear reminder The overall aim of PANFLUVAC is to construct measure. However, parenteral vaccination does of the need for better interpandemic as well as vaccine delivery systems for intranasal and not ensure induction of local immunity in the pandemic vaccines. parenteral vaccines. New H5N1 vaccines are to be respiratory tract — the route by which the virus based on well-established virosome technology infects humans — and from where it transmits The project allows for ‘robust scale-up of — proven its worth for efficacious interpandemic to other individuals. Inducing mucosal immunity vaccine production’, being coordinated by the vaccines — as well as whole virus vaccines. This by vaccination would enhance control of both leading group in virosomal technology – Crucell will permit comparison of the intranasal virosomal disease and transmission. Although mucosal (partner 2 CRU). CRU holds the intellectual vaccine with the whole virus vaccine. The vaccine immunisation has been studied, no acceptable property rights for the application of ISCOMs potency will be enhanced by novel adjuvants intranasal vaccine against influenza is yet for influenza vaccines. To this end, the project targeting dendritic cells, offering both antigen- available. A major problem for the intranasal employs the latest advances in reverse sparing potential and immunopotentiation vaccine has been the adjuvant, which is required (partner 3 - National Biological Standards Board) characteristics. Both ISCOMs and lipopeptide for efficient induction of immunity. — referred to in the Commission Working adjuvants are already proven immunopotentiators, Paper (COM(2004)201 final). This allows for biosafe for humans. Certain of these have been WHO recommendations novel vaccines to be provided within a short employed with experimental influenza vaccines The WHO assessment of the risk to human time. Vaccine efficacy will be enhanced through which allow the new H5N1 vaccines to be health from the H5N1 avian influenza virus application of novel formulations and adjuvants, fast-tracked in their development. Accordingly, states: 'The disease in humans has no vaccine in response to the requirement in the call for PANFLUVAC will generate the first H5N1 vaccine to confer protection and no specific treatment antigen-sparing strategies and the application of within the first 18 months of the project. once illness becomes severe.' The WHO warning novel immunopotentiating agents. The proposed that 'outbreaks in birds pose a significant threat adjuvants, both current and under development, The PANFLUVAC project is also designed to to human health', and that H5N1 'has the will provide the formulated vaccine with a potent facilitate rapid modification of the vaccine in potential to ignite a global influenza pandemic . Moreover, these selected the face of virus drift. Within the preclinical in humans', has led to an urgent requirement for adjuvants target the dendritic cells, which are evaluation, the new vaccines will be tested for the H5N1 pandemic influenza virus vaccines. The critically important for the activation of an degree of heterotypic cross protection they offer. PANFLUVAC vaccines fit this requirement and efficacious immune response. This allows for the PANFLUVAC offers a generic vaccine development allow for a rapid response to the entry of H5N1 development of more efficacious vaccination and system to provide safe and efficacious vaccines into Europe, enforcing the WHO recommendation better usage of the vaccine available. against influenza, fitting in with the European that 'trials of experimental influenza pandemic Commission’s Working Paper on Community vaccines for humans be accelerated'. By promoting the vaccine to reach the target Influenza Pandemic Preparedness and Response organs and cells more efficiently, lower doses of Planning. Antigen-sparing strategies and effects vaccine are required to maintain immunogenic of immunomodulatory molecules concentrations. 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Phase by assessed technologies existing from generated vaccine threat. influenza pandemic imminent of face the in vaccines required for need current evaluated. evaluations. pre-clinical in compared technologies) evaluation. preclinical for material and technologies existing from developed results pected : : applicable in a generic manner. The manner. The ageneric in applicable -are strategies evaluation and system delivery vaccine virosomal adjuvants, the - including employed being techniques the of Many P formulations are patented, while others developed developed others while patented, are formulations the of Certain vaccines. other to application for easily adapted be can protection of correlates vivo ex following: the are applications potential main the Overall, level. ahigh isat availability Property Intellectual and application patent project, the in SMEs of anumber of involvement the With application. patent to subject be will consortium the by g) Promoting human health and well-being, and and well-being, and health human Promoting g) for protection of correlates Immunological f) e) and mucosal enhancing adjuvants Safe d) vaccines. influenza for adjuvants as ISCOMs c) mucosal H5N1 for vaccine efficacious An b) parenteral H5N1 for vaccine efficacious An a) applications otential population confidence in the face of influenza. of face the in confidence population vaccines. efficacious new identifying rapidly experimentation. animal reducing evaluation, immunity. systemic administration. administration. In vitro In tests to be employed for immunological immunological for employed be to tests and and ex vivo vivo ex tests for vaccine vaccine for tests in vitro in : and and [email protected] UK 4NS E1 London Road End Ltd Mile 327 Virology Retroscreen O John [email protected] [email protected] UK 3QG Herts EN6 Bar Potters Mimms South Lane Blanche Virology of Division Control and Biological for Standards Institute National Wood John Prof. [email protected] Netherlands The Leiden 2301 V N Crucell Uytdehaag Fons Partners: admin.ch kenneth.mccullough@ivi. Switzerland Mittelhäusern 293 3147 (IVI) Sensemattstrasse and Immunoprophylaxis Virology of Institute D Coordinator: [email protected] [email protected] [email protected] Norway Hospital Bergen 5021 University Building Haukeland Hansen Armauer Institute Gade The and Immunology Microbiology of Dept Centre Influenza Bergen of University RHaaheim Lars Prof. r Kenneth McCullough r Kenneth xford Germany Braunschweig 78 38124 Inhoffenstrasse Guzman Carlos [email protected] 57/43 [email protected] 32 90 49 +39 Tel: Italy Rome 299 00161 Elena Regina Sanità di Viale Superiore Istituto Diseases and Immune-Mediated Parasite Infectious, of Centre Dept Influenza National D [email protected] 00 6269 44 Ext 00 82 20 +44 Tel: UK 5EQ NW9 London Avenue Colindale 61 Laboratory Health Public Central Zambon Maria [email protected] 261 [email protected] 30 69 21 +41 Tel: Switzerland Lausanne 1015 C PSE - Campus EPFL Scientifique Parc Sàrl, SCIPROM D [email protected] 12 99 [email protected] 17 +49 600 14 Mobile: 18 16 53 +49 Tel: r I r Kirsten Leufgen r Kirsten sabella D sabella onatelli

PANFLUVAC SP5B-CT-2007-044115 www.panfluvac.org Immunogenicity and protective efficacy of intranasal delNS1(H5N1) influenza vaccine

Acronym: Intranasal H5vaccine EC contribution: €2 680 400 Starting date: 01/01/2007 Key words: NS1, H5, avian influenza, vaccine, Duration: 36 months Instrument: STREP replication deficient, intranasal

Summary: Problem: Aim: In collaboration with six partners in four European Mankind is threatened by avian influenza. For The main objective of the current project aims to countries and in Russia, Green Hills Biotechnology the period of December 1, 2003, to February 3, define an intranasal H5N1 pandemic vaccine with is developing a novel vaccine against avian 2006, the WHO reported 161 confirmed human enhanced capacity to evoke a strong, long lasting influenza, within the framework of a research cases of avian influenza A (H5N1); of these 86 (or local and systemic immune response in humans. project subsidised by the European Union. The 53%) were fatal. It is not clear how many amino The replication-deficient vaccine is based project budget totals EUR 2.6 million. acid substitutions are still needed to convert avian on the deletion of the NS1 protein (DelNS1 viruses into a new human pandemic strain. Old vaccine). First, several vaccine candidates will This project is being carried out by an human influenza viruses of the H1N1 or H2N2 be screened for parameters including antigenic international consortium over a three-year period. subtypes could also return in humans, causing properties, receptor specificity, attenuation Besides Green Hills Biotechnology, a total of a new pandemic. Sooner or later there will be and immunogenicity in animal models. Studies three other companies and three universities are an influenza pandemic unless new and more on elucidation of mechanisms responsible participating in the development and evaluation efficient vaccines are developed. Therefore, the for protection will comprise parameters of an intranasal H5 vaccine. The proposed vaccine current situation with the H5N1 avian flu could be of innate and adaptive immunity. Several is based on proprietary technology from Green considered as a challenge for global science and standardised immunological techniques such as Hills Biotechnology. Immunological properties will the biotech industry. haemagglutination inhibition (HI) and ELISA will be assessed by the Goethe University in Frankfurt be used for evaluation of vaccine candidates. and preclinical experiments will be carried out In addition, novel sensitive methods based at Biotest s.r.o. in the Czech Republic. Novel, on synthetic sialic receptor analogues will be sensitive methods developed for assessment developed for assessment of the of the antibody response against viral proteins response against the haemagglutinin (HA) and are being established in cooperation with the neuraminidase (NA). Shemyakin & Ovchinnikov Institute of Bioorganic Chemistry in Moscow. These assays will allow the selection of an optimal vaccine candidate that will be subsequently evaluated in clinical trials at the Medical University, Vienna, and at Retroscreen in the UK.

10 Expected results: Potential applications: html Protective properties of the vaccine against The proposed clinical evaluation of delNS1 (H5N1) . homologous and heterologous influenza strains vaccine should be considered as a component of will be tested in ferret challenge experiments European systemic efforts to prevent and control using different H5 strains. To close the gap potential pandemics of avian influenza. This is a between the ferret model and human studies severe respiratory infection that poses a major

we will use the macaque model to evaluate the health threat on a worldwide scale. The proposed projects immune response. These pre-clinical studies vaccine will reduce mortality and morbidity rates, _ will allow the selection of the most potent lost workdays, and hospitalisations in case of eu vaccine candidate, which will be produced H5N1 outbreak. / according to cGMP guidelines on Vero cells. After toxicological evaluation of the vaccine, com clinical Phase I/II studies will be performed. If . permitted by authorities, a human challenge study using attenuated H5 virus will be carried out. Alternatively, for proof of principle of the delNS technology, an H1-delNS1 vaccine will be produced and used in an H1 challenge study. greenhillsbiotech . www

Partners: Ivana Šurová BioTest Ltd SP5B-CT-2007-044512 Pod Zámkem 279 281 25 Konárovice Czech Republic [email protected] Volker Wacheck Medical University, Vienna Währinger Gürtel 18-20 Vienna Austria [email protected] Jindrich Cinatl Clinics of J W Goethe- University Paul-Ehrlich-Str 40 vaccine 60596 Frankfurt Germany [email protected] Nicolai Bovin Shemyakin Institute of Bioorganic Chemistry Miklukho-Maklaya 16/10 117997 Moscow Coordinator: Russia Joachim Seipelt [email protected] AVIR Green Hills Biotechnology John Oxford AG Retroscreen Virology Ltd Gersthofer Strasse 29-31 327 Mile End Road 1180 Vienna London E1 4NS Austria UK [email protected] [email protected] ntranasal H5 I Dose sparing and increased immunogenicity for vaccination against pandemic influenza with CoVaccine HT

Acronym: FluVac Key words: Adjuvant, vaccine, dose-sparing, EC contribution: €3 500 000 Starting date: Planned for 01/07/2007 efficacy, influenza, pandemic, emergency Duration: 48 months Instrument:STREP vaccination

Summary: Problem: An adjuvant may compensate the shortcomings The risk of a new influenza pandemic is emphasised The impact of an influenza pandemic can be of the seasonal influenza vaccines for application by a WHO report documenting several hundreds of enormous as illustrated by the ‘Spanish Flu’ during a pandemic. Firstly, an adjuvant may recent cases of human infection with a new virus from 1918 to 1919 - one of the most dramatic reduce considerably that antigen dose required to strain and approximately 50% mortality. Control or events in human history with 20 to 40 million establish a certain level of immunity. Secondly, prevention of a pandemic by emergency vaccination casualties worldwide. Influenza pandemics were it may have a beneficial effect on the type, level depends on availability of products with high caused by influenza virus A subtypes. Wild birds and kinetic of the immune response. Thirdly, it efficacy and broad protection. The combination of as the natural hosts can easily spread the virus may decrease the number of low-/non-responders an effective adjuvant and cell culture technologies and infect other animals and humans. Recently, thereby affording a higher degree of immunity in for antigen production meets the requirements of a a new highly pathogenic H5N1 strain of avian a susceptible population. Fourthly, it may enhance pandemic influenza vaccine. influenza A virus has become epidemic in birds in cross-protection to distinct influenza virus Asia and has been transmitted to European and strains. Feasibility studies with CoVaccine HT, a The consortium of this FluVac project, among them African countries. Mortality rates among infected sucrose fatty acid sulphate ester incorporated proven and renowned experts in human vaccine humans exceeded 50%, which is alarming. This in a submicron emulsion of squalane-in-water development and influenza research, plans to deliver H5N1 virus did not spread between humans, but exerted the ability to induce strong humoral and a prototype vaccine with proof-of-concept within 48 once an influenza virus acquires the ability to cellular immune responses against a wide range months. Optimal doses of inactivated, cell culture- transmit through respiratory droplets from one of antigens in various animal species and high derived whole influenza virus (H5N1) and CoVaccine human to another it could spread within a few efficacy in combination with low antigen dose HT as adjuvant will be tested in pre-clinical and months around the world. The consequences of (Fig. 1). clinical studies. In comparison to the widely used a pandemic outbreak could be enormous as it production system in eggs, production in cell culture could disrupt daily life with tremendous social and The combination of these technologies opens offers the advantages of immediate and reliable economic effects and put a large burden on the unique opportunities for preventing and availability of antigen of constant quality and with system. High costs of anti-influenza controlling an influenza pandemic and the reduced risk of contamination. drugs and limited availability thereof, implicates knowledge and expertise gained might be an urgent need for an affordable vaccine for the applicable to the design of other (types of) The use of whole virus in a vaccine instead of prevention and control of a pandemic influenza vaccines. subunits, is considered to induce broader protective outbreak. immunity. The novel adjuvant CoVaccine HT has Aim: been shown to elicit high humoral and cellular Currently, most human influenza vaccines are The consortium aims at a novel influenza responses against different types of antigens, produced in embryonated chicken eggs, which vaccine formulation by combining a novel including inactivated influenza virus, and in different demands strict logistics. In case of a pandemic adjuvant CoVaccine HT and an inactivated, cell animal species (mice, rats, rabbits, pigs, horses, influenza outbreak, the availability of eggs might culture-derived, whole influenza virus (H5N1). primates, etc.). It is considered to be a promising be a critical factor. Therefore, this project exploits Feasibility studies with the adjuvant indicated candidate for a pandemic influenza vaccine. A cell culture technology for antigen production that CoVaccine HT is a promising candidate for Phase I trial, human challenge study and extensive as this method is more flexible, independent of emergency vaccines to establish high levels of immunological evaluation of the vaccine prototype supply of animal-derived materials and more immunity and to compensate for the limited will provide the information required for further consistent. With a master and working cell- availability of antigen. development and registration (Scheme 1). bank system, production and up-scaling can be immediately started in case of emergency. The ultimate goal of the FluVac project is to prove safety and efficacy of a CoVaccine HT adjuvanted Schematic representation of the programme. pandemic whole H5N1 virus vaccine in humans Explorative studies Pre-clinical studies Clinical studies and to gain insight in its performance in animal Immunogenicity (adult, neonatal, GMP manufacturing Phase I trial Safety Immunogenicity elderly mice) Validation methods Optimisation Human Challenge study models. After extensive preclinical evaluations, formulation a vaccine with the optimal composition will be Efficacy (ferrets) (ferrets, rabbits) studied in a Phase I and subsequently in a human challenge study. In parallel, the Extensive immunological characterisation of humoral and cellular responses 12 vaccines will be investigated in mouse models for 100,000

10,000

1,000

A number of defined objectives will be pursued The consortium envisages that this project 100 Anti-HINIneonates and Anti-HINIelderly. The ambitiousAnti-H3N2 preclinicalAnti-H3N2 during the proposed project: will contribute to the urgent need of improved antibodyand clinicaltitle developmentantibody title planantibody addresses title all antibody title adjuvants for emergency vaccines. Knowledge knownPV1 aspects ofPV2 protective immunityPV1 against PV2 a) Delivery of non-GMP and cGMP grade and expertise gained will be useful to other types influenza. CoVaccine HT and H5N1 antigen for use in of vaccines. Commercial influenza vaccine preclinical, and clinical studies. b) Evaluation of dose sparing approaches Commercial influenza vaccine by incorporating CoVaccine HT, by diluted 1/10 in PBS Commercial influenza vaccine analyses of humoral and cellular diluted 1/10 in CoVaccine HT responses in mice and ferrets. c) Proof of efficacy of the prototype 100,000 vaccine in protection models in ferrets with current pandemic influenza virus strain(s). 10,000 d) Determination of kinetic and quality of humoral, cellular and mucosal immune responses especially in high-risk 1,000 populations in mouse models for elderly and neonates. e) Development of a stable vaccine using 100 validated methods. Anti-HINI Anti-HINI Anti-H3N2 Anti-H3N2 antibody title antibody title antibody title antibody title f) Achievement of pre-clinical proof of PV1 PV2 PV1 PV2 safety in animal toxicology studies. g) Evaluation of safety and Effect of 1/10 dilutionCommercial in PBS influenza or CoVaccine vaccine HT of a commercial influenza vaccineCommercial on ELISA antibody influenza titres vaccine in pigs (data kindly immunogenicity in human clinical Phase I trial. provided by CoVaccinediluted 1/10BV). in PBS h) Evaluation of protection and immunogenicity Commercial influenza vaccine diluted 1/10 in CoVaccine HT in a human challenge study induced by the CoVaccine HT-adjuvanted pandemic influenza Expected results: vaccine. Successful completion of the FluVac project and subsequent development and registration of the vaccine will have great benefits for the health of Potential applications: the European population. In addition, the project The FluVac consortium proposes a novel product

will contribute to the development of a novel for (pre-)pandemic vaccination by combining 44407 generation of adjuvants and of improved vaccines Protherics’ proprietary adjuvant CoVaccine HT to combat infectious diseases. The consortium is with OrganonBiosciences’ H5N1 viral antigen supported by the complementary know-how and produced in cell culture. Upon successful technology of individual partners. The participants completion of clinical proof-of-concept studies, are renowned international experts in such areas the consortium will pursue further clinical as human (influenza) vaccine development, development. adjuvant development, neonatal immunity and influenza research i.e. Protherics (United Currently there are no adjuvanted Influenza Kingdom), OrganonBiosciences (the Netherlands), vaccines licensed and marketed in the USA. MF59 Retroscreen (United Kingdom), Landspitali and virosomes are licensed in Europe for epidemic PARTNERS: Dr James Glover University Hospital (Iceland) and Erasmus Medical and Al(OH)3 for pandemic influenza vaccine. Protherics plc Centre (the Netherlands). [email protected] Prof. Dr Ingileif Jónsdóttir University of Iceland Landspitali University Hospital and Faculty of Medicine Dept of Immunology Hringbraut 101 Reykjavik Iceland [email protected] Prof. Dr John S Oxford Retroscreen Virology Ltd Centre for Infectious Diseases Bart’s and The London Queen Coordinator: Mary’s School of Medicine and Scientific Coordinator: Dentistry Dr Luuk A Th Hilgers 327 Mile End Road [email protected] London E1 4NS Project Manager: UK Dr Jacco G M Heldens [email protected] [email protected] Dr Guus Rimmelzwaan OrganonBiosciences / Nobilon Erasmus Medical Center International B.V. Dept of Virology P.O. Box 320 Dr Molewaterplein 50 ac Exportstraat 39b 3015 Rotterdam

5830 Boxmeer The Netherlands V The Netherlands [email protected] lu F Novel AI DIVA recombinant vaccines for duck

Acronym: NOVADUCK EC contribution: €1 416 380 Starting date: 01/01/2007 Key words: Avian influenza, recombinant vaccines, Duration: 36 months Instrument: STREP vectored vaccines, ducks, DIVA, immune response

Summary: Problem: Aim: The NOVADUCK project brings together both The ongoing outbreak of H5N1 HPAI has recently The NOVADUCK project aims to develop and private (two companies and one SME) and spread from Asia to Africa and Europe, posing evaluate new highly protective and cost-effective public sector stakeholders (European reference a real public threat as HPAI can occasionally avian influenza live vaccines for ducks based on laboratories and agencies on avian influenza) infect humans. Ducks play a major role in the live vectors and in line with the DIVA strategy. from four European countries with the aim of epidemiology of avian influenza because wild More specifically, the NOVADUCK project will: developing and evaluating new highly protective waterfowl, including ducks, constitute the natural and cost-effective avian influenza live vaccines for reservoir of all subtypes of influenza A virus. a) identify the optimal AI immunogenic sequence ducks, based on live viral vectors and in line with Experimental infection of ducks with recent to be inserted into the selected live vectors; the DIVA strategy (Differentiating Infected from isolates indicates a longer shedding period and a b) generate and optimise three types of live Vaccinated Animals). selection for lower virulence variants, suggesting recombinant vector-based vaccines; that duck has become the ‘Trojan horse’ of Asian c) develop reliable and cost-effective duck- Viral vectors will be engineered to optimise both H5N1 AI. specific immunological tools to measure the their immunogenicity and protective capacities. immune response induced by the different Vaccine candidates will be pre-screened for safety Although biosecurity is the first line of defence vaccine candidates and to detect infection in a and immunogenicity using newly developed against HPAI, strategic use of vaccination is vaccinated duck (DIVA strategy); duck-specific immunological tools. The best clearly recognised as a tool to help eradicate HPAI d) assess the safety and immunogenicity of the vaccine candidates will be evaluated for efficacy in an infected country. Most studies evaluating new vectored vaccine candidates and compare in an HPAI H5N1 challenge-model in ducks. A the efficacy of AI vaccines have been performed it with those of existing vaccines to select the serological DIVA test, able to detect infection in in chickens, and duck studies have been relatively best vaccine candidate(s); vaccinated duck flocks, will be generated, and the rare. Existing inactivated AI vaccines are less e) set up a challenge model in ducks for vaccine effect of vaccination on genetic and antigenic drift immunogenic in ducks than in chickens and evaluation of efficacy; of H5N1 will be assessed. must generally be administered twice to be fully f) measure the efficacy of the most immunogenic efficient; furthermore, there is no commercially vectored vaccine candidates against recent available DIVA test to monitor AI infection in HPAI H5N1 and compare it with existing birds injected with this type of vaccine. Therefore, vaccines; highly efficient, cost-effective, DIVA-compatible g) study the effect of vaccination on genetic/ AI vaccines for ducks are still greatly needed. antigenic drift; h) select the best candidate(s) to be developed In this specific context, live vector-based vaccines based on its (their) immunogenic and protective hold the greatest promise and are one of the most properties as well as its (their) estimated cost of effective options. Indeed, some live recombinant production and administration mode flexibility vector-based AI vaccines have shown excellent (e.g. individual versus mass administration). results in chickens, but they are not necessarily adapted for use in ducks. Expected advantages of this type of vaccine include administration at Expected results: a younger age, mass administration, rapid onset The NOVADUCK project is designed to identify of immunity, and compatibility with the DIVA the optimal AI immunogenic sequence to be strategy. inserted into a vector. New specific tools to evaluate humoral, cellular and mucosal immunity The NOVADUCK project is designed to induced in ducks will be developed. Three types demonstrate and exploit the potential of live of vector-based vaccines containing the optimal vector vaccines to develop a new generation of immunogenic sequence will be generated, highly efficient and cost-effective AI vaccines and their immunogenicity using different for ducks and therefore could contribute to administration modes will be compared to that of eradicating AI from the ecosystem. existing inactivated AI vaccines using the newly 14 developed immunological tools. A challenge model using the most recent and representative Potential applications: an infected area, or for preventive vaccination in AI isolates in which clinical signs, oral and cloacal The newly developed vector-based AI vaccines for high risk or enzootic area. shedding can be significantly detected in all ducks generated during the NOVADUCK project challenged birds will be set up in ducks and will could be available for vaccination of ducks in Some of the newly developed vaccine candidates be used to compare the efficacy induced by the countries in which the biosecurity measures are may also be safe and efficacious in other species, newly selected vector-based vaccine candidates not sufficient to control AI H5 infection. The including other poultry (e.g. chickens, turkeys with that induced by existing vaccines. Studies will selected vaccine candidates should show clear and geese), but also mammals (e.g. cats, dogs, be designed to verify that the newly developed advantages over existing vaccines, such as DIVA- pigs, horses), making them ideal multi-species vaccines show clear advantages over existing compatibility, low cost, early onset of immunity, AI vaccine candidates. Additionally, if efficacy vaccines. The compatibility of these vector- adjuvant-free, and/or ease of administration (e.g. can be obtained after administration by the oral based vaccines with newly developed DIVA tests mass vaccination, duckling administration at the route, the potential exists to use such vaccines will also be verified. In addition, data will be hatchery). These new types of vaccines would to vaccinate wild ducks and thus potentially generated on the effect of vaccination of ducks on be suitable for emergency vaccination, thereby contribute to the eradication of AI from the genetic/antigenic drift of the challenge virus. limiting the mass slaughtering of poultry around ecosystem.

WP00 – Management activities (MERIAL) The duck-specific immunological tools that will be developed during the NOVADUCK project would also be essential to evaluate the immunogenicity WP01 – Preparatory Phase (CVI) of any existing, as well as future, duck vaccines. In particular, the DIVA test could be used to monitor infection in vaccinated flocks.

WP02 – MERIAL The development of a reproducible AI challenge Generation and production of optimised viral vectored vaccines for ducks (MERIAL) model in ducks will be useful to define the standard for evaluation of efficacy of duck AI vaccine. The protection data generated during WP04 – Safety assessment and the project would also help to define the minimal WP03 – Development of pre-screening of Immunological tools (VAR) vaccine candidates (VMRI) level of protection that should be expected for an acceptable AI duck vaccine. Overall, the project should give new tools to control AI infection in a species playing a significant role in AI epidemiology. WP05 – Protection studies (vaccination-challenge) and drift (AFSSA)

Research Activities

WP06 – Innovation activities and societal aspects (MERIAL) SSPE-CT-2006-044217

Innovation Activities

Management Activities NOVADUCK PROJECT STRUCTURE

Partners: Dr Csaba Dren Virology Dept - Veterinary Dr Véronique Jestin Veterinary Medical Research Laboratories Agency Agence Française de Sécurité Institute of the HAS Woodham Lane Sanitaire des Aliments Avian Immunology and Tumour New Haw Laboratoire d’Etudes et Virology Addlestone KT15 3NB de Recherches Avicoles et Hungária krt 21 UK Porcines P.O. Box 1581 Pf 18 Tel: +44 19 32 35 73 39 BP53 Zoopôle 1143 Budapest [email protected] 22440 Ploufragan Hungary Fabienne Mathieu France Tel: +36 14 67 40 98 Biosource Europe Tel: +33 29 60 16 222 [email protected] R&D Hybridoma Dept [email protected] Dr Vilmos Palfi Rue de l’Industrie 8 Dr Thierry Vandenberg Central Veterinary Institute 1400 Nivelles Veterinary and Agrochemical Dept of Virology Belgium Research Centre (VAR) Tábornok utca 2 Tel: +32 67 88 99 49 Coordinator: Dept of Small Stock 1149 Budapest [email protected] Dr Michel Bublot Diseases, Avian Virology and Hungary Charlotte Dalba Merial SAS Immunology Unit Tel: +36 14 60 63 29 Epixis S A 254 Rue M Mérieux Groeselenbergstraat 99 [email protected] 16-18 Rue de la Glacière 69007 Lyon 1180 Brussels Dr Ian Brown 75013 Paris France Belgium The Secretary of State For France Tel: +33 47 27 25 973 Tel: +32 23 79 06 30 Environment, Food and Rural Tel: +33 14 21 76 520 [email protected] [email protected] Affairs [email protected] NOVADUCK Vaccine, diagnostic test development and immunology aspects of avian influenza

Acronym: AIV VACC DIAGNOSIS Key words: Avian influenza viruses, Newcastle EC contribution: €1 372 890 Starting date: 01/12/2006 disease viruses, vaccines, diagnostics, Duration: 36 months Instrument: STREP immunology, DIVA

Summary: of infected from vaccinated animals (DIVA In order to better control avian influenza, more Avian influenza is a zoonotic disease and is seen principle). The development of sensitive, specific effective mass applicable vaccines are needed. as one of the most important emerging diseases and easy to use marker diagnostic tests that will For better design of future vaccines and strategies with serious economic consequences. Although be compatible with the vaccines is another goal of to combat avian influenza more insight is also some vaccines for poultry are available, all this project. required into the immunological mechanisms vaccines have considerable drawbacks with regard and characterisation of immune responses after to dose and application methods (injection), onset Problem: vaccination and infection. of immunity, efficacy or costs of production and The highly pathogenic H5N1 avian influenza An easy to use, sensitive and specific serological application which limit their use. (AI) currently circulating in Asia, and recently in test allowing the DIVA principle, which can be northern and western Africa and Europe, has used in conjunction with the use of inactivated or The primary aim of this project is to develop better led to the deaths of more than 150 million birds vectored vaccines, is needed. avian influenza vaccines through live or vector and over 150 humans. Due to the seriousness vaccines that could be mass applicable through of this threat, some countries are taking steps spray, drinking water or eye drop. These vector to vaccinate their entire poultry population. Aim: vaccines would offer considerable advantages Currently, a consensus is emerging that An ideal vaccine to be added in controlling - mass applicable, less labour intensive and vaccination of birds at risk could be a critical AI should be: i) efficacious in reducing virus animal friendly application, protection by local part of a control strategy in averting a human transmission; ii) genetically close to the circulating and systemic immunity and less interference with pandemic. virus; iii) serologically distinguishable from wild eventual maternal , more complete type virus; iv) applicable by mass administration protection through cellular and humoral immunity, Although very useful in the fight against avian routes; and v) inexpensive. faster onset of immunity when used in face of an influenza, all currently available influenza vaccines outbreak and cheaper production methods. have considerable shortcomings; several vaccines Recent achievements showed that safe and developed over the past two decades to protect efficacious NDV vectors could be constructed The project exploits recently acquired knowledge poultry against the highly pathogenic H5 or H7 that would make development of a vaccine with concerning the molecular characterisation of are based on inactivated whole virus vaccines. the above characteristics possible. NDV vectors the viruses resulting in the construction of Apart from the challenge of setting up a robust carrying H7 respectively H5 gene inserts were candidate strains with highly interesting efficacy diagnostic test for differentiating vaccinated recently constructed and 100% protection against and safety profile. Safety and efficacy with from infected animals, these vaccines have to be clinical disease as well as considerable reduction Newcastle disease (NDV) vectors and infectious administered by labour intensive and expensive in virus shedding was observed after challenge laryngotracheitis (ILT) vectors both for H5 and parenteral injections. with NDV and AI. However, the H7 and H5 ORF for H7 inserts have already been demonstrated used for the construction of these vectors was in vivo. In view of the worrying spread of epidemic not derived from recent isolates and it can be avian influenza H5N1 and the large undertaking expected that protection will be enhanced when A system in which gene cassettes for the foreign to vaccinate billions of birds in some parts of using vector vaccines carrying recent H5 or H7 proteins can easily be constructed and exchanged the world, development of efficacious vaccines genes. This knowledge will be used to construct will be developed and will be able to respond that could be administered by mass application optimised NDV vector vaccines that will have very quickly to a change in antigenicity of the routes, such as spray or drinking water, is urgently advantages in terms of their improved efficacy field virus. Further optimised additional candidate needed. In the endeavour to develop improved against recent H5N1 isolates. The NDV parent strains will be constructed and extensively vaccine against AI, recombinant DNA technology strains that will be used for the construction tested. Experiments on genetic in vitro and in was employed to generate vectored, subunit or of the recombinants are a vaccine strain that is vivo stability, immunological responses, virulence DNA vaccines. Although a wide range of these widely used in the field for years and with a well testing, spreading, and transmission studies in vaccines has been experimentally shown to be established safety record. chickens, ducks and other avian species will be effective against AI, only a fowl pox-vectored performed. vaccine with H5 gene insert is commercially Objectives of the vector vaccines constructed in available. This recombinant vaccine, however, also the project: The vaccines to be developed would also have requires administration by parenteral injection. a) H5 genes genetically close to the currently 16 marker aspects which will allow the differentiation circulating high pathogenic Asian H5N1 viruses. com

b) Safe and highly efficacious against lethal Potential applications: . challenge with recent highly pathogenic H5N1 Live vaccines with much easier methods of strains. Protection against clinical disease and application will result in less labour and thus reduction in excretion of the challenge virus. lower costs of vaccination. Methods of application c) Earlier onset of immunity and protection, via natural route (drinking water, spray, eye drops) because of local protection as compared to the are easier to perform and in certain areas could currently available inactivated vaccines. offer considerable advantages over parenteral diagnosis d) Administration via mass applicable methods vaccination. Less costly and easier vaccination - such as drinking water, spray or eye drop. methods will also increase compliance with e) Serologically distinguishable from wild type virus vaccination campaigns and policies, especially in vacc

by use of an easy-to-use accompanying Elisa. developing countries. -

Vaccination by more natural application routes aiv

Expected results: . This project is expected to deliver the first mass will also result in a much more animal friendly applicable live vaccine against highly pathogenic procedure, as stress due to vaccination will be H5N1 avian influenza. The vaccine would offer considerably reduced and animal welfare greatly major technical advantages, including production improved. The project will also deliver fast, www aspects, over the currently existing vaccines. sensitive, robust and specific ELISA tests. This will enable the DIVA concept. These tests can be used An easy to use, sensitive and specific serological test in conjunction with the newly developed vaccines allowing the DIVA principle, which can be used in or with existing vaccines. conjunction with the use of inactivated or vectored vaccines, will considerably enhance optimal avian The availability of easy to perform, fast and influenza vaccination and control strategies. reliable test systems that could be used for mass- testing is a prerequisite for the development and Better knowledge of the critical pathways involved successful implementation of control measures in influenza immunity and immunopathology may and eradication policies. eventually contribute to improved vaccine design, and optimised immunisation or other intervention The successful use of the live vaccine and DIVA strategies. tests will offer major possibilities and tools in the Partners: Dr Thomas Mettenleiter fight against avian influenza and thus will have a Friedrich-Loeffler-Institut The development of molecular biological tools major contribution to economical poultry farming Federal Research Institute for Animal Health will allow in the future a quicker response to and human health. Boddenblick 5a changes in antigenicity of the field-virus. The 17493 Greifswald-Insel Riems Germany construction of gene cassettes that will allow fast Coordinator: [email protected] integration of the H region of future genetically Dr Danny Goovaerts Tel: +49 38 35 17 250 and antigenically different influenza strains in the Intervet International bv Dr Christian Schelp Wim de Körverstraat 35 Dr Bommeli AG

ND vector will allow a much faster response for P. O. Box 31 Stationsstrasse 12 SSPE-CT-2007-044141 construction of updated vaccine strains. 5830 AA Boxmeer 3097 Leibefeld-Bern The Netherlands Switzerland Tel: +31 48 55 87 727 Tel: +41 31 97 06 265 [email protected] [email protected] AIV VACC DIAGNOSIS VACC AIV Development of a combined influenza/SARS vaccine

Acronym: SARS/FLU VACCINE EC contribution: €1 607 500 Starting date: 01/01/2005 Duration: 36 months Instrument: STREP Key words: Vaccine, SARS, influenza

Summary: Problem: Aim: The spontaneous appearance of SARS in humans In 2002, an atypical pneumonia, characterised The project uses Green Hills’ engineered in 2003 and the regular yearly appearance of by progressive respiratory failure, emerged in influenza virus as its starting point. This modified influenza provoke an obvious question: could it be the Guangdong Province in Southern China. The virus is unable to replicate, but still expresses possible to produce a single vaccine that protects causative agent was rapidly identified as a new immunogenic influenza proteins on its surface. against both diseases? coronavirus which was designated as severe acute The sequences for relevant antigens are identified respiratory syndrome-associated virus SARS-CoV. by bioinformatic methods and were verified by A project led by an Austrian SME aims to do The disease swept rapidly to neighbouring regions immunological methods. Promising antigens from exactly this. Green Hills Biotechnology is well and led to several cases even in Toronto, Canada. the SARS-associated coronavirus will be expressed acquainted with influenza: it currently has several By the end of the epidemic in July 2003, about by the influenza vector so that it expresses SARS products in development based on genetically 8 000 SARS cases and almost 800 deaths due to proteins too. Several constructs are tested for modified versions of the virus. SARS had been recorded worldwide. Since then, immunogenicity: the one that provokes the best the world is in an inter-epidemic period, as no immune response without compromising safety The company is now using its expertise in viral new cases were reported. in animal testing is selected for a full preclinical engineering to develop a combined SARS/flu testing programme. vaccine. It has teamed up with three other SMEs (in Austria, the Czech Republic and Slovenia) which have added their expertise in antigen identification, preclinical testing and protein purification. The project also includes two renowned university research institutions that bring additional expertise in virology and immunology.

18 immunisation. one with virus influenza and SARS-CoV against responses immune protective induce to potential the has it that advantage major the has project this in developed as vaccine CoV. aSARS/flu Such SARS- the as such pathogens foreign of antigens of expression the for avector as candidate attractive an also but virus influenza against vaccine attenuated live asafe as attractive only –make not it properties IFN-inducing its to due immunogenic highly being while replication abortive to due –apathogenicity virus influenza modified the of properties particular The Ex results pected : other relevant pathogens as well. well. as pathogens relevant other to applied be to potential the have will project this in used are that technologies The immunisation. one with virus influenza and SARS-CoV against concept immunisation an towards step major a be will project this in developed vaccine The P applications otential : Paul-Ehrlich-Str 40 Paul-Ehrlich-Str Goethe-University W J of Clinics Cinatl Jindrich Partners: [email protected] Austria Vienna 29-31 1180 Strasse Gersthofer Biotechnology AG Hills Green AVIR T Coordinator: [email protected] Austria Vienna 5/3 1010 Rathausstrasse GmbH Biodevelopment Emergentec Mayer Bernd [email protected] Slovenia Ljubljana 1000 30 d.o.o. Teslova Separations BIA A [email protected] Germany Frankfurt 60596 homas Muster homas leš Štrancar leš [email protected] Vienna 1030 9/3 Vienna Bohrgasse Dr University, Medical S Joachim [email protected] Republic Czech Konárovice 28125 279 Zámkem Pod Ltd BioTest I vana Šurová Šurová vana eipelt

SARS/FLU VACCINE LSHB-CT-2004-512054 www.greenhillsbiotech.com/eu_projects.html Novel antigen-adjuvant vehicle as an effective influenza vaccine

Acronym: Universal Vaccine EC contribution: €1 154 717 Starting date: 01/06/2005 Key words: Antigen-adjuvant vehicle, universal Duration: 24 months Instrument: SMEs - Cooperative Research Projects influenza vaccine, mucosal immunity

Summary: The unique combination of the consortium for the Aim: One of the biggest challenges concerning rational design of a mucosal influenza vaccine is The Universal Vaccine project strives to meet this influenza vaccination is trying to keep up with unprecedented in European vaccine research. If medical need by employing a highly innovative the virus’s mutational variation. The currently the universal vaccine proves successful in clinical strategy that represents a conceptually novel approved vaccines work by stimulating the trials it will not only help to diminish the social way of rationally designing a vaccine against body’s immunity against the haemagglutinin and and economic costs of influenza, but also secure influenza. The aim of the Universal Vaccine neuraminidase proteins on the virus’s surface. As the growth and development of the European project is to develop a powerful, new, safe and these proteins are prone to mutation, vaccines vaccine industry in the global market. easily-administered nasal vaccine for humans that only induce immunity against specific subtypes of provides lifelong protection against influenza. the virus. However, the influenza virus has a third Problem: protein in its outer coat, M2, and the extracellular Influenza is a recurrent global threat and The scientific excellence and technologies within domain of this protein, M2e, has been remarkably affects millions of people in the world every the consortium will be combined to design, conserved in the amino acid sequence since year. Vaccination constitutes undoubtedly the develop, formulate and evaluate novel M2e- human influenza virus was first isolated in 1933. If most cost-effective preventive measure against based influenza vaccine candidates with the aim this protein could stimulate an adequate immune morbidity and death from infectious diseases of achieving improved efficacy, longer lasting response it might be possible to develop a broad- such as influenza. Current influenza vaccines protection and broad-spectrum immunity against spectrum vaccine against all influenza A subtypes. are based on the major influenza glycoproteins the diversity of influenza strains, exceeding the haemagglutinin (HA) and neuraminidase (NA) performance of current annual influenza vaccines. Previous research has shown that when the as antigenic determinants. However, these In the longer perspective the project may help extracellular domain of M2 (M2e) is linked to proteins are subject to mutation (drift) and reduce or even eradicate influenza infections in appropriate carrier particles, such as the hepatitis gene re-assortment (shift) generating seasonal humans. B virus core, it becomes highly immunogenic, variations in the circulating influenza strains inducing antibodies that fully protect mice against thereby rendering this type of vaccine less a potentially lethal influenza infection. Swedish efficient in long term protection against viral biotech company Biovitrum AB has teamed up infection and necessitating annual updates of the with researchers at the Flanders Interuniversity vaccine components. Furthermore, most vaccines Institute for Biotechnology in Ghent (BE), who today are injectable and use aluminium salts initially worked on the M2 protein, European as their adjuvant component. However, there SMEs Pepscan (the Netherlands), Proxima (UK) is general agreement that needle-free vaccines and Göteborg University to develop what could are preferable for reasons of improved patient become the first universal vaccine for influenza. It comfort and ease of administration, as well as could provide lifelong immunity against the virus reducing the risk of contamination and other and thus provide far greater protection in the adverse effects while promoting compliance and event of a pandemic. It may even help to eradicate increasing the safety of vaccination. Moreover, for the disease in humans. respiratory diseases like influenza, nasal delivery brings the vaccine to where it is most needed to Another feature of this vaccine is its nasal stimulate a local immune response, namely the administration. For respiratory diseases like respiratory tract. influenza it is beneficial to bring the vaccine to where it is most needed to stimulate a local immune response, and nasal vaccination may help to induce stronger or more lasting immunity in recipients. Furthermore, needle-free nasal sprays are safer and easier to administer, reduce the risk of contamination – and are far less likely to deter people from participating in vaccination 20 programmes. org Expected results: Potential applications: . The development of a universal influenza vaccine The Universal Vaccine project will play a is based on the identification of an extracellular pivotal role in the future development and domain of the minor influenza protein M2 protein, competitiveness of the SMEs involved. A referred to as M2e. The sequence of the M2e successful novel mucosal vaccine against domain has been highly conserved since the influenza would have significant impact on first isolation of human influenza virus in 1933, the global market and secure growth and Coordinator: Dr Anna Ramne despite numerous epidemics and at least two development of the European vaccine industry. Biovitrum AB major pandemics. Therefore, by utilising the M2e The results of the project will be investigated BioTech Huset Arvid Wallgrens Backe 20 peptide as antigenic determinant a vaccine will be for patentable knowledge and it is anticipated 41346 Göteborg created that could provide long term protection that the SMEs could establish themselves firmly Sweden [email protected] against a broader spectrum of influenza strains. on the market for mucosal vaccines. Their universalvaccine . extended and potentially stronger IP would allow Partners: The development of needle-free vaccines has expansion of their respective business areas and Dr Hans Langedijk Pepscan Systems BV been hampered by the lack of effective mucosal open up new opportunities for exploitation and Edelhertweg 15 adjuvants. toxin (CT) and the closely commercialisation. 8219 Lelystad www The Netherlands related Escherichia coli heat-labile toxin (LT) are [email protected] powerful mucosal adjuvants but have proved Dr Roger New unsuitable in the clinical setting due to their Proxima Concepts Ltd The Biosciences Innovation inherent toxicity and potential association with Centre Bell’s palsy (paralysis of the facial nerve). A novel Royal Veterinary College Royal College Street mucosal adjuvant, CTA1-DD, has been developed London NW1 which combines the enzymatic activity of the UK [email protected] A1 unit of cholera toxin with a dimer of the Ig- Dr Doriano Cingolani binding element of Staphylococcus aureus Protein Eurogentec SA A, thereby targeting the adjuvant specifically to LIEGE Science Park Rue Bois Saint-Jean 5 B-cells and other Ig-binding cells of the immune 4102 Seraing system. This strategy has proved successful in Belgium [email protected] that the CTA1-DD adjuvant is completely non- Prof. Walter Fiers/Dr toxic and, despite its more selective binding Xavier Saelens Flanders Interuniversity properties, has retained potent mucosal and Institute for Biotechnology systemic immunoenhancing functions making it a VZW (VIB) highly suitable adjuvant for development of new, Dept for Molecular Biomedical Research DMBR Dr Nils Lycke efficient mucosal vaccines. Ghent University Göteborg University (UGOT) VIB FSVM-building Dept of Microbiology & Technologiepark 927 Immunology The operational goals of the project are to: 9052 Ghent Medicinaregatan 7A a) covalently fuse the M2e-peptide with the Zwijnaarde Box 435 CTA1-DD adjuvant, creating a strongly Belgium 40530 Göteborg

[email protected] Sweden COOP-CT-2005-017749 immunogenic influenza vaccine suitable for [email protected] [email protected] mucosal delivery; b) further improve vaccine efficacy by incorporating the fusion protein in proprietary liposomes; c) increase the in vivo maintenance of the antigen by using blocked or constrained peptides or peptidomimetics; d) determine the in vivo mechanism of action of the mucosal influenza vaccine; e) demonstrate the safety and efficacy of the vaccine in animal challenge models.

For reasons of safety, efficacy and cost, mucosal administration of vaccines is today a priority for immunising against mucosal as well as systemic

infections. The ease of administration of needle- accine free vaccines facilitates their distribution to larger populations, particularly in the developing world, while improving vaccination safety and compliance. This will have great positive impact on global health and immense societal gains. The long term effect of a conceptually novel influenza vaccine will contribute to reducing and perhaps even eradicating influenza infections in humans. V niversal U Generation of information and tools to support the management of the avian influenza crisis in poultry

Acronym: FLUAID EC contribution: €1 200 000 Starting date: 01/01/2006 Duration: 30 months Instrument: STREP Key words: Avian influenza, vaccines, diagnostics

Summary: Problem: Aim: Avian influenza (AI) has become a great risk Avian influenza outbreaks have recently The primary goal of this proposal will be the both for animal and human health. In five caused severe losses to the poultry industry, its joint development and application of novel years, over 200 million birds have been affected stakeholders and ultimately to the EU taxpayer. technologies to combat AI infections. This by this disease, including estimations of the It is estimated that since 2000, 200 million goal will be achieved through the interaction ongoing H5N1 epidemic. By bringing together birds have died or have been culled following of leading European institutes along with the both European and non-European laboratories, infection with influenza viruses subtypes H5 active collaboration of laboratories in Indonesia, the FLUAID project aims to generate data or H7. Approximately 50 million of these birds Pakistan, South Africa, Thailand and Vietnam. on significant issues linked to AI outbreak were from Europe. Most importantly, human management, about which scientific knowledge is infections have also been reported in several of currently lacking. these outbreaks. The ongoing H5N1 outbreaks are a serious concern for food security and human health on a global level, with the crossing of the species barrier representing a serious potential risk of a new human pandemic virus emerging. The increased relevance of AI in the fields of animal and human health, has highlighted the lack of scientific information on several aspects of the disease. This has hampered the adequate management of some of the recent crises thus resulting in millions of dead animals and concern over loss of human lives and over management of the pandemic potential.

22 diagnosis of AI. of diagnosis rapid and vaccination to related be will project this during developed applications primary The P research. further and making decision support to used be will and visibility gain will achievements scientific and knowledge experience, European way this In globally. applied be to disease the for measures control effective more of development the to contribute will virus, AI the of transmission and pathogenesis on knowledge improved with coupled This, partners. European non- and European both by validated be will which tests pen-side sensitive and rapid novel, develop also will It test. diagnostic companion avalidated and bank EU an vaccine for strains candidate including vaccination, of use the for guidelines specific make available will FLUAID Ex applications otential results pected : : D Coordinator: [email protected] [email protected] Netherlands The Lelystad (CIDC-Lelystad) Health Science and Animal for Institute D [email protected] UK 3NB Surrey KT15 Addlestone Lane Woodham Dept Agency Virology Laboratories Veterinary D Partners: [email protected] Italy (PD) Venezie Legnaro delle Sperimentale Zooprofilattico Istituto Newcastle and Disease for Influenza Avian Laboratory Reference National and OIE\FAO r I r Guus Koch r Guus Banks r Jill laria Capua Capua laria [email protected] 293 Mittelhaeusern Sensemattstrasse and Immunoprophylaxis Virology of Institute D [email protected] Thailand Bangkok 10440 Rajathevi Road Payathai 69/1 D [email protected] Pakistan Islamabad Road Park National Agricultural Centre Research D [email protected] France Ploufragan 22440 53 B.P. Ploufragan de site - AFSSA Aliments des Sécurité de Sanitaire Française Agence D r Kenneth McCullough r Kenneth Buranathai r Chantanee N r Khalid Jestin r Veronique aeem [email protected] Africa South Stellenbosch 7602 St Victoria 1 X1 Box Stellenbosch of University Biochemistry of Dept D Prof. [email protected] France Montpellier 340810 Valsiére la de Vet Rue 1682 Diagnostic Innovative Pourquier Philippe Mr [email protected] UK 1LZ YO41 York Hutton Sand Laboratory Science Central D Chris Mr [email protected] Australia Road Geelong Portarlington 5 24 BAG O. P. Health Laboratory Animal Industries Australian Livestock CSIRO D r Paul S r Paul irk Uwe Bellstedt Uwe irk elleck anks [email protected] Vietnam Hanoi Dong-da, St Giai-Phong Health Lane 11-78th Animal of Department Veterinary Diagnosis for Center National D Indonesia Yogyakarta 27 Km Wates Wates – Yogya Centre Raya JL Investigation Health Disease Animal of Directorate D r T r I sep S sep hanh Long T Long hanh ulaiman o

FLUAID SSPE-CT-2005-022417 www.fluaid.eu Live attenuated replication-defective influenza vaccine

Acronym: FLUVACC EC contribution: €9 200 000 Starting date: 01/09/2005 Key words: Replication, deficient, vaccine, Duration: 60 months Instrument: Integrated Project influenza

Summary: Problem: Aim: Green Hills Biotechnology is developing a novel Industrial production of influenza vaccine still FLUVACC has improved its core technology for vaccine against influenza in a joint project that relies on traditional techniques. Essentially, live attenuated vaccine production, using a brings together the expertise of eight different chicken eggs are used as mini vaccine factories. technique called reverse genetics. Together with partner institutions both from academia and They are injected with live influenza virus and the project partners, Green Hills Biotechnology biotech industry in four European countries incubated for several days so the virus can has developed a ‘master strain’ that is lacking and in Russia. The ambitious project is being multiply. The egg is then opened, the virus the NS1 gene which is essential for productive carried out by the international consortium over harvested, purified and inactivated. Unfortunately, viral replication. Candidate vaccines for emerging a five-year period. The FLUVACC vaccine is a highly pathogenic avian viruses do not grow well influenza subtypes can be quickly produced by novel component of European systemic efforts in eggs as they tend to kill the embryo. inserting their genes into this master strain so that to prevent and control influenza, based on a they express the immunogenic surface proteins, replication deficient virus that is generated by a There are many other problems associated with but remain replication-deficient. This master specialised technique called reverse genetics. The egg-based production. The whole process is time strain was adapted to grow to high titers in vaccine will be produced in cell culture. intensive and hard to scale up, so that during a tissue culture, making it possible to produce large pandemic it may be difficult for supply to meet quantities in the case of a pandemic. demand. In addition, the combination of vaccine with egg proteins can lead to allergic reactions in some people.

FLUVACC aims to shift vaccine production away from the traditional methods by generation of live attenuated-replication deficient vaccines that can be produced in cell culture. Instead of using egg-produced viral proteins, the live attenuated vaccines developed by the FLUVACC consortium contain whole replication deficient viruses that generate a strong immune response but are non- pathogenic.

Medical University Vienna BIA Separations d.o.o. Michael Bergmann Coordinator: Aleš Štrancar Währinger Gürtel 18-20 Joachim Seipelt Teslova 30 1090 Vienna AVIR Green Hills Biotechnology 1000 Ljubljana Austria AG Slovenia Michael.bergmann@ Gersthofer Strasse 29-31 [email protected] meduniwien.ac.at 1180 Vienna BioTest Ltd Robert Koch Institute Austria Martin Šlais Thorsten Wolff [email protected] Pod Zámkem 279 Nordufer 20 281 25 Konárovice 13353 Berlin-Wedding Czech Republic Germany Partners: [email protected] [email protected] GPC Biotech AG Russian Academy of Weikom & Network – Bert Klebl Medical Sciences Agency for Communication Fraunhoferstrasse 20 Oleg I Kiselev Maria Weidinger-Moser 82152 Martinsried 15/17 Prof Popova Str Gilgegasse 11/16 Munich St. Petersburg 197376 1090 Vienna Germany Russian Federation Austria 24 [email protected] [email protected] [email protected] hospitalisations. and workdays, lost rates, morbidity and mortality reduce will vaccine proposed The scale. worldwide a on threat health amajor poses that infection respiratory isasevere Influenza influenza. control and prevent to efforts systemic European of FLUVACCcomponent anovel The isas vaccine P 2007. of half first the in performed be will Istudies Phase Clinical developed. was Vero on cells based process production GMP a and ferrets, and mice in evaluated were candidates vaccine The influenza. pandemic and endemic for strains candidate vaccine several produced and evaluated developed, have FLUVACC the partners project this With Ex applications otential results pected : :

FLUVACC LSHB-CT-2005-518281 www.greenhillsbiotech.com/eu_projects.html Development of influenza del NS1 virus as a vector for foreign antigens

Acronym: CHIMERIC VACCINES Starting date: 01/11/2004 EC contribution: €1 384 945 Instrument: SMEs – Co-operative Key words: Influenza, avian, vaccine, adjuvant, Duration: 30 months Research Projects chimeric

Summary: Problem: Aim: The project CHIMERIC VACCINES is being carried Avian influenza is an infectious disease of birds This consortium has developed a novel approach out by an international consortium over a 30- caused by type A strains of the influenza virus. for vaccination against avian influenza. The month period. The consortium aims at developing The disease occurs worldwide. While all birds are technology is based on the insertion of selected a novel vaccine that will provide protection thought to be susceptible to infection with avian epitopes into a genetically modified influenza against avian influenza and seasonal influenza. influenza viruses, many wild bird species carry virus that is apathogenic. To achieve these results, The technical basis is a replication deficient these viruses with no apparent signs of harm. both a stable vector and promising antigens influenza virus that was modified to express A highly pathogenic form was first identified in were identified. Antigen selection was based on foreign antigens. Italy in 1878 and can lead to a mortality rate in bioinformatics methods that were substantiated infected animals of 100% within 48 hours. by experimental validation. The properties of the CHIMERIC VACCINES is led by the Austrian SME backbone vector in terms of safety and stability Green Hills Biotechnology and consists of three Influenza viruses are species-specific and only were assessed in preclinical experiments and gave partner companies in Austria, Slovenia and the rarely cause infection in other species. Since highly satisfactory results. To bring the proposed Czech Republic and two renowned university 1959, instances of human infection with an avian chimeric vaccine into clinical trials, a production partners in Austria and Germany. In order to bring influenza virus have been documented on only process in small scale was established using a these vaccines to clinical use, one of the partners 10 occasions. Of the hundreds of strains of avian novel purification technology. Finally, preparations is the Russian WHO reference laboratory for influenza A viruses, only four are known to have for a clinical trial in Russia were made by the influenza in St Petersburg, which has abundant caused human infections: H5N1, H7N3, H7N7 Russian Institute of Influenza. experience in testing new vaccines. and H9N2. In general, human infection with these viruses has resulted in mild symptoms and very little severe illness, with one notable exception - the highly pathogenic H5N1 virus.

Currently, the H5N1 virus is of greatest significance as it has crossed the species barrier to infect humans on at least three occasions in recent years leading to the current outbreaks that Partners: began in December 2003. Currently, no vaccine Emergentec against avian influenza is available. Biodevelopment GmbH Rathausstrasse 5/3 1010 Vienna Austria BioTest Ltd Pod Zámkem 279 281 25 Konárovice Czech Republic BIA Separations d.o.o. Teslova 30 1000 Ljubljana Slovenia Clinics of J W Goethe- University Paul-Ehrlich Str 40 60596 Frankfurt Germany Medical University, Vienna Coordinator: Dr Bohrgasse 9/3 Joachim Seipelt 1030 Vienna Avir Green Hills Biotechnology Austria AG State Institution Research Gersthofer Str 29-31 Institute of Influenza 1180 Vienna Prof Popov Street 15/17 Austria P. O. Box 197376 j.seipelt@greenhillsbiotech. St Petersburg 26 com Russian Federation production of purified viruses will be established. established. be will viruses purified of production the for aprocess and generated be will safety and immunogenicity regarding information important addition, In vaccine. achimeric of concept the develop further to help will and strategies vaccination novel of development the in aspects important on light shed will project this of Results Ex results pected : other relevant pathogens as well. well. as pathogens relevant other to applied be to potential the have will project this in used are that technologies The pathogens. of antigens foreign against response immune an induce can that avaccine towards step important an be will project this in developed vaccine The P applications otential :

CHIMERIC VACCINES COOP-CT-2004-512864 www.greenhillsbiotech.com/eu_projects.html Novel vaccination strategies and vaccine formulations for epidemic and pandemic influenza control

Acronym: NOVAFLU Key words: Influenza, vaccine, strain selection, EC contribution: €1 679 211 Starting date: 01/10/2002 computer modelling, cell culture, MVA, ISCOM, Duration: 36 months Instrument: Shared Cost Action defective viruses, CTL, animal model

Summary: vaccination. The major drawbacks of the currently genetics technology which allows the genetic Annual influenza epidemics and pandemics cause used multi-component influenza vaccines, manipulation of viruses. It was the objective to a significant disease burden and death rates produced in embryonated chicken eggs, are their engineer influenza viruses and develop reverse in human beings, and have a high economical limited efficacy and classical production methods genetics technology that would allow the rapid impact on the EU. The major drawbacks of which are hard to adapt to requirements related generation of reassortant strains that give high influenza vaccines are their limited efficacy and to ‘drift’ and ‘shift’ phenomena. yields of the relevant viral proteins, the HA and production methods that are hard to adapt to the NA. requirements of continuously changing influenza viruses. We aim to improve the efficacy and Aim: In Work Packages 2 and 3 the development of production methods of epidemic and pandemic The aim of the project was the development of these reverse genetic systems was addressed. influenza vaccines through: more effective strategies for the vaccination of For the production of viruses in embryonated human beings against epidemic and pandemic chicken eggs or chicken embryo fibroblasts, a) Improving epidemic vaccine strain selection influenza. Divided over the respective work the avian POL1 promoter was cloned which based on retrospective and prospective packages four objectives were identified. These can drive the transcription of viral RNA in analyses. were: avian cells. It was not possible to optimise b) Improving vaccine strain selection and the extremities of the HA gene segments reference reagents for pandemic influenza. a) Development of optimal strategies for for enhanced vaccine production yield since c) Implementation of reverse genetics vaccine selection: Computer algorithms were with a reporter gene expression system it technologies to increase flexibility and yield of developed which are used for the interpretation was demonstrated that these non-coding influenza vaccines. of complex serological data and allow the regions are already compatible with the RNA d) Use of cell substrate for vaccine production. easy assessment of antigenic relatedness complexes of various viruses including that e) Use of novel antigen delivery systems. of influenza viruses. These tools have been of the vaccine back-bone strain A/PR/8/34. f) Detailed examination of correlates of implemented and fully integrated in the bi- Furthermore, a universal reverse genetics protection. Promising strategies were tested in annual WHO vaccine strain selection process. system was developed based on the T7 appropriate animal models. In addition, these newly developed tools can polymerase and the T7 promotor system. This be used for the antigenic characterisation of would allow the transcription of RNA in any cell the potentially pandemic H5N1 strains. Thus type independent of the species from which the Problem: in the case of a pandemic outbreak, the prior cells originated (e.g. MDCK cells). Annually occurring influenza virus epidemics, knowledge of the antigenic properties of associated with antigenic ‘drift’ of the surface these viruses can aid in the selection of the A patent application was filed on the T7 glycoproteins haemagglutinin and neuraminidase best vaccine strains to provide the broadest reverse genetics system. The availability of an (HA and NA) of the influenza A and B viruses, protection possible. This scenario was exercised alternative reverse genetic system constitutes cause a significant disease burden and high with the avian H7N7 viruses that caused an a very valuable alternative that would prevent death rates in human beings which exceed deaths outbreak of fowl plague in the Netherlands in the use of existing systems and for which the due, for example, to road accidents in Europe. 2003 (see c). Extensive surveillance of avian intellectual property rights outside the EU. Less frequent pandemic outbreaks of influenza influenza viruses amongst wild birds resulted In addition the use of the T7 system is better (three in the last century) associated with the in a library of virtually all subtypes of HA and tailored for the use in MDCK cells to which EU introduction of a new influenza A virus subtype NA, which were used for vaccine preparation companies have access. from an animal reservoir - antigenic ‘shift’ - are and the preparation of reference reagents to even more serious in this respect, causing millions aid in the adequate diagnosis and identification c) Development of novel vaccine candidates: This of deaths in a short period of time. of avian influenza virus infections in man and was addressed in three different ways. First, animals. recombinant MVA vectors were developed Although recently a new generation of antiviral and evaluated as a novel delivery system compounds (neuraminidase inhibitors) with b) Development of alternative approaches for for influenza viral proteins. MVA vectors promising anti-viral effects have been developed, vaccine production with the recent registration were constructed that express the HA of the most cost-effective and efficient way of MDCK-cells as an influenza vaccine two different strains of H5N1 virus. These 28 of controlling influenza remains preventive production platform and the advent of reverse recombinant H5N1 poxvirus vector vaccines d) Definition of correlates of protection: For the the For protection: of correlates of Definition d) induced against the lethal challenge of H5N1 of challenge lethal the against induced be could immunity protective preparations vaccine these with that demonstrated was it and model amouse in evaluated were observed with a mutation in an epitope and and epitope an in amutation with observed was co-mutations of anumber case one In virus. the to isadvantageous variation this that indicating significantly, response CTL the affected epitope an in change acid amino asingle that found was it and response CTL human specific virus the on studied was epitopes individual in variation of impact The infections. virus influenza of control the in correlates immune important are CTL that evidence is indirect which pressure, selective under are CTL human by recognised epitopes that observed was it First, response. CTL human the and viruses influenza between interaction the of made was analysis Adetailed (CTL). T lymphocytes cytotoxic specific virus on put was emphasis of alot protection of correlates of determination immunity. mediated cell- also but responses antibody induce only not will it that isexpected it since promising, isconsidered particles virus defective on based Avaccine procedure. this on filed was application apatent and complementation trans- by particles defective produce to possible proved It replication. its for isessential that gene functional one lack that produced were Viruses particles. virus defective of production the for exploited was technology genetics reverse Again, particles. virus influenza defective of use isthe vaccines candidate novel of development the for approach third The virus. H7N7 alethal with infection against protected were and antibodies specific virus developed which mice, of immunisation the for used and ISCOMS with adjuvanted was preparation This NA. and HA antigens viral the of production the for prepared was strain H7N7 vaccine an procedures selection strain vaccine novel and surveillance influenza avian technology, genetics of reverse a Using combination system. delivery antigen alternative an as (ISCOMS) complexes stimulating immune of use the was evaluated was that approach second The influenza. of outbreaks pandemic of face the in shortage vaccine envisaged the overcoming to contribute can capacity, production vaccine influenza existing of is independent production the which of candidates, vaccine MVA recombinant that isanticipated It viruses. H5N1 the including viruses, influenza various of gene (NP) nucleoprotein the expressing MVA constructed were Furthermore, filed. was application apatent data these on Based virus. Tel: +31 10 40 88 066 [email protected] 88 40 10 +31 Tel: Netherlands The Rotterdam 50 3015 Molewaterplein Dr Virology of Dept MC Erasmus D Prof. Coordinator: mutations associated with escape from CTL CTL from escape with associated mutations point of examples more some and assessed was epitopes CTL of variation of extent the Also, epitope. the in substitution acid amino the of effect detrimental the for compensate functionally to necessary were mutations co- the that found was It epitope. the in substitution asingle with technology genetics reverse by virus rescue to impossible proved it pandemic strains of influenza. of strains pandemic against also immunity, broad-spectrum of induction the for approach afeasible be may epitopes conserved these against immunity Tcell of induction the that indicate may This fitness. viral of loss the without substitutions acid amino introduce to impossible proved it analysis amutational In constraints. functional under are epitopes conserved certain that found also was it response, CTL specific virus of induction the at aim that vaccines of development the complicate could epitopes CTL of variation the Although identified. were r ADMEO sterhaus Tel: +44 20 78 82 79 66 79 [email protected] 78 20 +44 Tel: UK 4NS E1 London Road End Mile 327 Ltd Retroscreen D Prof. 40 21 [email protected] 77 03 61 +49 Tel: Germany Langen 51-59 63225 Paul-Ehrlich-Strasse Paul-Ehrlich-Institut D Prof. 722 [email protected] 88 56 14 +33 Tel: France Paris Roux 75724 Docteur du Rue 28 Pasteur Institute D Prof. 542 [email protected] 77 44 29 +31 Tel: Netherlands The Weesp 36 1380 Bv Houtenlaan van J C Pharmaceutical Solvay S 66 44 [email protected] 33 23 12 +44 Tel: UK 3EJ CB2 Cambridge Street Downing Cambridge of University D Partners: jirk Kok jirk r DJS r JO r GS Werf der r Svan mith xford utter P filed. were applications patent three and project the from emanated journals peer-reviewed international in publications 75 of Atotal 91 on occasions. conferences international and national at presented were project the during obtained Results Ex e) The induction of virus-specific CTL may be a be may CTL virus-specific of induction The e) (MVA, systems delivery antigen Novel d) used be can technology genetics reverse Novel c) for used be could subtypes HA/NA of Alibrary b) antigenic the for algorithm computer The a) applications otential way to prepare broadly protective vaccines. vaccines. protective broadly prepare to way pandemic. a of face the in vaccines of shortage envisaged the overcoming in aid may and vaccines effective of production the for used be can ISCOMS) of use and particles virus defective CEF cells. and cells MDCK in strains vaccine of preparation the for vaccines. prototype of preparation the selection. strain vaccines pandemic for used be could and process selection strain vaccine influenza annual the in applied isalready viruses influenza of characterisation results pected : :

NOVAFLU QLK2-CT-2002-01034 Preparing for an influenza pandemic

Acronym: FLUPAN EC contribution: €2 100 000 Starting date: 01/12/2001 Key words: Influenza, pandemic, vaccine, bird flu, Duration: 36 months extended to 48 months Instrument: Shared-Cost Action reverse genetics

Summary: Status (January 2007): Aim: Although the threat of avian influenza has only A vaccine candidate against the H7N1 bird flu To diagnose more effectively, the emergence and recently made major media headlines, Europe virus has been developed using reverse genetics spread of potential pandemic influenza viruses began developing its defences many years ago. technology and egg-free vaccine production from animals to man and to be able to respond In September 2001, a team of scientists from the technology. This new vaccine, called RD-3, went rapidly by producing safe, effective vaccines. UK, Italy and Norway collaborated with vaccine into pre-clinical immunogenicity studies in early researchers from Sanofi Pasteur in France on 2006, and safety/efficacy Phase I clinical trial the FLUPAN project. Funded through the Fifth with 60 volunteers at the end of 2006. The Framework Programme, the project aimed to protocols and techniques for monitoring human develop a candidate vaccine for human pandemic infections with avian influenza and carrying out flu. clinical trials, which the FLUPAN consortium has developed will be important tools in pandemic The partners decided to target the H7N1 avian preparedness and will provide methods that can influenza subtype which caused lethal outbreaks be used to develop vaccines also against H5N1 in in Italian poultry in 1999. In 2003, 80 people in the event of a pandemic. the Netherlands were infected with the related H7N7 subtype caught from poultry; one person died of the disease. Problem: At the start of the FLUPAN project, we were Although recent media attention has been ill-prepared to react to the emergence of highly devoted to the H5N1 subtype, researchers believe pathogenic avian influenza viruses in man. Coordinator: that H7 subtypes could also cause a pandemic. Reverse genetics technology had never before Dr John Wood National Institute for Biological The deadly H7N1 strain is too dangerous for been used to produce influenza vaccine viruses Standards & Control standard influenza vaccine production, so the and there was little experience in producing and Blanche Lane South Mimms FLUPAN scientists used a technique called reverse testing potential pandemic vaccines. It was vital to Potters Bar EN6 3QG genetics to alter the H7 protein and make the rehearse our pandemic preparedness by involving Hertfordshire virus safe. This process also modified the virus so not only research scientists and vaccine specialists UK [email protected] that it could be grown in a mammalian cell line as but also health and safety regulators and vaccine well as the more usual poultry eggs. The use of a licensing agencies. Partners: Dr Fred Vogel mammalian host for the virus makes large-scale Sanofi Pasteur production of a human vaccine easier and safer. 1541 Avenue Marcel Mérieux 69280 Marcy I’Etoile The resulting vaccine will be the first influenza France pandemic vaccine produced entirely in mammalian [email protected] cells. Dr Isabella Donatelli Istituto Superiore di Sanita Dept of Infectious, Parasitic But the FLUPAN project has done more than and Immune-Mediated Diseases Dr Wendy Barclay produce a potential vaccine against H7 avian Viale Regina Elena 299 University of Reading influenza. In other strands of this research, 00161 Rome School of Biological Sciences Italy P.O. Box 228 surveillance of avian influenza viruses in Italy [email protected] WhiteKnights has enabled the partners to build up a library of Prof. Lars Haaheim Reading RG6 6AJ reagents which will be a valuable resource for University of Bergen UK Influenza Centre [email protected] pandemic vaccine development in the future; Dept of Microbiology and Prof. Maria Zambon new tests to monitor antibodies induced by avian Immunology Central Public Health The Gade Institute Laboratory influenza viruses have been developed which Armauer Hansen Building Respiratory Virus Unit should improve our ability to detect emergence of Haukeland University Hospital 61 Colindale Avenue 5021 Bergen London NW9 5EQ new pandemic viruses. Norway UK 30 [email protected] [email protected] /

Expected results: Potential applications: a) Improved understanding of influenza viruses As the tools and techniques developed by FLUPAN circulating in domestic and wild animals in the can be adapted to develop new vaccines quickly fluplan

EU. and efficiently, for example against H5N1, the / b) Use of reverse genetics to derive a safe project has made an important contribution to vaccine virus from a highly pathogenic H7N1 Europe’s pandemic preparedness. The experience avian virus. gained in developing safe working practices c) Production of cell culture H7N1 vaccine for for genetic modification of highly pathogenic preclinical and clinical trial. influenza viruses and in devising a testing protocol

d) Demonstration of H7N1 vaccine for demonstrating safety of vaccine viruses spotlight / immunogenicity and efficacy in mouse model. derived by reverse genetics has been invaluable

e) Demonstration of H7N1 vaccine safety and in producing WHO biosafety risk assessments for uk . immunogenicity in a Phase I/II clinical trial. H5N1 vaccine development.

f) Development and validation of new H7N1 ac . antibody tests. nibsc . www QLK2-CT-2001-01786 FLUPAN Prove the mucosal adjuvanticity of LT mutants with influenza antigens for intranasal immunisation

Acronym: MUCADJ Key words: Control of infectious diseases, pre- EC contribution: €1 003 359 Starting date: 01/02/2000 clinical development of vaccines, transdisease Duration: 36 months Instrument: Shared Cost Action vaccinology

Summary: Problem: Aim: Mucosal administration of antigens is a global Delivery of vaccines by mucosal route is one of the This project was officially initiated on February priority. Many approaches have been adopted to major goals of today’s research. The advantage 1, 2000, and concluded its technical and develop vaccines that can be delivered without of easier delivery combined with the possibility experimental activities on January 31, 2003. using syringes and that induce immune response of neutralising pathogens at their portal of entry at the mucosal sites that are often the portal have made mucosal immunisation one of the The key objectives of the project were to of entry of pathogens. Here we propose to major targets of the European Union, WHO and demonstrate: manufacture GMP lots of the mucosal adjuvants the US NIH. Furthermore, mucosal delivery by a) that the genetically detoxified mutants of LTK63 and LTR72, two non-toxic derivatives eliminating the use of syringes would increase E. coli enterotoxin act as mucosal adjuvants of E. coli enterotoxin obtained by site-directed compliance and decrease the risk of spread of and can be safely administered to humans by mutagenesis, and to test them in a clinical setting infectious diseases that has been reported by intranasal route; in human adult volunteers to obtain proof of improper use of syringes during vaccination. b) that subunit influenza antigens formulated concept that these molecules can be effectively with the LT mutants induce an increased used to adjuvant mucosal vaccines for human use. In spite of the obvious advantages offered by immune response at the mucosal sites As a model, for this demonstration project we mucosal vaccination, and the success obtained and a systemic response similar to that have chosen the influenza vaccine for intranasal in this area in animal models in recent years, of conventional vaccines for parenteral administration. mucosal vaccines are not yet a reality, and it administration; remains unclear whether they can be used for c) the role in humans, if any, of appropriate human vaccination. It is therefore mandatory delivery systems to increase the to obtain proof of concept of the feasibility in immunogenicity of intranasally administered flu humans of mucosal vaccines. vaccines adjuvanted with LT mutants.

Most vaccines are unable to induce an immune response when delivered at mucosal sites. In order to make them immunogenic, strong mucosal adjuvants are required. In animals, the most potent mucosal adjuvants are the E. coli enterotoxin (LT) and cholera toxin (CT). Unfortunately, these two toxins cannot be used in humans and therefore their use as mucosal adjuvants has been restricted to animal studies.

Recently, by site-directed mutagenesis, mutants of LT have been obtained which are either completely non-toxic or have greatly reduced toxicity. Preclinical studies have shown that these mutants still act as mucosal adjuvants when used in animal models, and that the LT mutants are generally more active than the CT mutants. Two particular LT mutants, LTK63 and LTR72, both of which are completely non-toxic or with very low residual activity showed, respectively, a good safety and adjuvanticity profile in preclinical studies, suggesting that they could be proposed for human studies. 32 mucosal level. mucosal the at response immune the enhances it addition in and humans in administration intranasal for adjuvant amucosal as used safely be LTK63 can that shown indeed have studies our Overall, R d) The demonstration in the proof of concept concept of proof the in demonstration The d) and safety of demonstration The c) in potency and safety of demonstration The b) vaccines influenza LTK63 The adjuvanted a) been have milestones project following The esults achieved: in humans. humans. in vaccines available currently to compared level mucosal the at response immune the enhance vaccines influenza mucosal that trial clinical trial. clinical concept of proof hoc ad an of means by achieved been has humans in administration intranasal for vaccines influenza mucosal of immunogenicity species. animal different in studies immunogenicity and toxicological hoc ad of means by achieved been has formulations vaccine these of model animal the developed. been have administration, intranasal for system, delivery without and with formulations, : of the MUCADJ project will also be used for the the for used be also will project MUCADJ the of course the in produced results and knowledge The E. in coli. produced LT mutants recombinant by targeted be could which diarrhoea, traveller’s against vaccines of development the including vaccines, mucosal other many of development clinical the to way the open potentially to useful be will project this from deriving results The P future pandemics. future for vaccines of design rational the for implications important have findings these whole, the On papers: following the in published been already have project MUCADJ the in obtained results clinical and preclinical key .The MF59TM emulsion adjuvant the with combined candidate vaccine leading as virus (H5N3) A/Duck/Singapore/97 influenza variant anon-pathogenic using participants MUCADJ most by started been already have studies several To H5N1 purpose, the this as such influenza. pandemics future for vaccines of development • Stephenson I, Zambon M, Rudin A, Colegate Colegate A, Rudin M, Zambon I, • Stephenson P, Ruggiero S, Morant M, • Peppoloni Contorni applications otential 80 (10): 4962-4970. (10): 80 Vol 2006; Virology of Journal volunteers. adult using adjuvants, mucosal as biovector novel and enterotoxin coli Escherichia non-toxigenic with vaccine influenza inactivated trivalent intranasal of IEvaluation Phase K. Nicholson K, J,Mills B,Holmgren Bonnington A, Minutello G, Giudice Del R, Bugarini A, Podda A, 293. 285- 2(2), 2003, Vaccines. Rev. Expert vaccines. of delivery intranasal for adjuvants strong and safe as enterotoxin labile heat- coli Escherichia the of Mutants G. Giudice Del Aand Podda R, Rappuoli MG, Pizza M, : Tel: +44 11 62 58 61 64 61 [email protected] 58 62 11 +44 Tel: UK 5WW LEI Leicester Square Infirmary Leicester of University D Partners: 96 34 [email protected] 24 77 05 +39 Tel: Italy Siena 53100 1 Fiorentina Via Srl and Diagnostics Vaccines Novartis D c Tel: +39 02 23 61 387 [email protected] 61 23 02 +39 Tel: Italy Milan 38 20133 Pascal Carlo Via Virologia Ist Milan of University D 34 41 [email protected] 23 77 05 +39 Tel: Italy Siena 53100 1 Moro Aldo Via Igiene Ist Siena of University D oordinator: oordinator: r Karl GN r Karl r A r Fabrizio Pregliasco r Fabrizio r E manuele Montomoli manuele udino Podda udino icholson

Tel: +35 31 60 83 57 38 57 83 60 31 +35 Tel: Ireland 2 Dublin Biochemistry of Dept College Trinity D Tel: +44 18 12 00 44 00 44 [email protected] 00 12 18 +44 Tel: UK 5HT NW9 London Ave Lab Health Colindale 61 Public Central PHLS D 911 24 34 31 [email protected] +46 Telephone: Sweden Goteborg 10 41346 Microbiol Guldhedsgatan Medical of Dept Goteborg of University D [email protected] r Kingston HGMills r Kingston r Maria Zambon r Maria Holmgren r Jan

MUCADJ QLK2-CT-1999-00070 www.altaweb.it/mucadj Combating flu in a combined action between industry and the public sector in order to secure adequate and fast intervention in Europe Acronym: FLUSECURE Key words: Pandemic influenza, public-private EC contribution: €3 749 685 Starting date: 01/02/2006 partnership, cross protection, reassortants, Duration: 36 months Instrument: Public Health Programme adjuvants, vaccine efficacy, clinical network

Summary: Problem: Aim: New or deliberately released viruses are a Pandemic influenza currently is the greatest a) To enable the production of the most effective continuing serious health threat for which no infectious diseases threat to public health. pandemic vaccine in the shortest possible time effective countermeasures are available. The The establishment of a public consortium is in sufficient quantity for the EU population. main problem is that no vaccine can be produced essential for assisting manufacturers in the timely in advance as the exact nature of the virus is production and testing of influenza vaccines for b) To establish a European network of public not known until the outbreak starts. Pandemic the European population. health institutes for a public-private partnership influenza is now felt to be the major health on European pandemic influenza vaccines. threat, especially with respect to the difficulties in effective preparedness planning. As pandemic c) To enhance vaccine preparedness in the case influenza is a cross-border problem, it needs a of an influenza pandemic. cross-border solution. This requires that European industry and public health authorities collaborate to gain control over it. As an overall objective this project aims to ‘enable the manufacture of the most effective pandemic vaccine in the shortest possible time in sufficient quantity for the EU population, by setting up a European network of public health institutes as the public sector input for a public-private partnership‘.

34 eu . Expected results: Potential applications: a) The establishment of a process of continuous a) Creation of a library of tested reassortants dialogue with vaccine manufacturers to ensure and reagents which will be available for direct that the assets of the public and private sectors vaccine production. are used to the best advantage to increase

the availability of vaccines to the European b) Optimised vaccine formulation for influenza flusecure population. vaccines in terms of formulation, adjuvants and . dosing regimen. b) A decrease in the lead time of vaccine

production by establishment of a library of c) Quantification efficacy assays for cross www safety-tested reference influenza strains protection and advanced correlates of available to industry. protection.

c) An improvement in the efficacy of pandemic d) Multi-centre clinical trial network and post influenza vaccines in terms of formulation, marketing surveillance network available for adjuvants and dosing regimens. pandemic vaccine studies in Europe. Provides a platform for public (governments and d) Quantification of vaccine efficacy by academia) and private (vaccine manufacturers) establishment of cross protection and advanced studies on pandemic vaccines. correlates of protection.

e) Evaluation of the pandemic vaccines in use by providing a strategy to set up a multi-centre clinical trial network and post marketing surveillance. This will assist industry in producing efficacy and surveillance data in the event that vaccines are produced and utilised under the mock-dossier scheme. 2005207

©Robert Koch Institut, Germany

Partners: Health Protection Agency UK www.hpa.org.uk Institute Pasteur France www.pasteur.fr Norwegian Institute of Public Health Norway www.fhi.no National Institute of Public Health Finland www.ktl.fi Institut Za Varovanje ZdravjaInstitute of Public Health of the Republic of Slovenia Slovenia www.ivz.si Statens Serum Institut Coordinator: Denmark Prof. Dr B A M van der www.ssi.dk Zeijst Cantacuzino Institute Project Leader: Romania Dr E D L Schmidt www.cantacuzino.ro/en Nederlands Vaccin Instituut (NVI) Robert Koch Institute Antonie van Leeuwenhoeklaan 11 Germany P.O. Box 457 www.rki.de 3720 AL Bilthoven National Centre for The Netherlands Epidemiology [email protected] Hungary www.nvi-vaccin.nl www.epidemiologia.hu/oek FLUSECURE Diagnostics and Surveillance

36 Development and standardisation of methods to detect and identify influenza viruses are essential to ensure the efficient diagnosis of influenza and to monitor spread of the virus in animal and human populations. This knowledge is, in turn, essential for the earliest possible intervention steps in control of the disease.

A broad range of different diagnostic tools is already available, ranging from direct antigen-detection and molecular assays such as (real-time) PCR to a variety of different serological tests that detect the response of the infected host. Two important problems that need to be tackled by researchers, however, are a) the adaptation of these methods to the strain H5N1, which has pandemic potential and is not well detected by several routine methods and b) the robustness and high-throughput use of these tests in ‘field’ or point-of-care conditions. These problems apply to both the animal and human health field. While during a true pandemic, health services are likely to have to rely on clinical diagnosis alone (also called ‘syndromic triggering’), robust diagnostic tests are crucial in the early phase of a pandemic for surveillance purposes.

Mapping the spread of an epizootic (epidemic in animals) is a key part of targeting control measures such as culling or vaccination of animals as well as specific awareness-raising and training of the population in the affected area. Similarly, epidemiological surveillance in humans is a prerequisite not only to monitor potential outbreaks, but also as a basis for a number of important research questions, such as the seasonality of ‘normal’ influenza epidemics or its spread into different population and age groups. Sophisticated multi-parameter modelling becomes increasingly important in linking existing data and developing different scenarios for future outbreaks.

The vast majority of projects funded to date in the field of diagnostics and surveillance originate in the animal health community – and it will be the task of future calls for proposals to fill the respective gaps in the area of human health: NEW-FLUBIRD unites the expertise of virologists and ornithological organisations worldwide to study the contribution of migratory birds to the spread of avian influenza. FLURESIST and RIVERS examine the key public health question of how long influenza viruses can survive and be detected in animal carcasses, commodities and the environment. FLUTEST, LAB-ON- SITE (which also considers a number of other animal diseases) and AVIFLU (which also includes vaccine research) all focus on the development of sensitive and robust diagnostic tests. ESNIP and ESNIP2 are making important contributions to the diagnosis and surveillance of influenza infections in pigs – taking into account their role as potential ‘mixing vessels’ for human and avian viruses. The European Influenza Surveillance Scheme (EISS), funded through the Public Health Programme, has established an important Europe- wide surveillance network for human influenza infections. Improved diagnosis and early warning systems for avian influenza outbreak management

Acronym: FLUTEST EC contribution: €1 502 880 Starting date: 01/02/2007 Key words: Avian influenza, poultry, highly Duration: 36 months Instrument: STREP pathogenic, diagnostic

Summary: Problem: Aim: The primary goal of this proposal will be the joint In recent years, Avian influenza (AI) outbreaks This project aims to generate data on significant development and application of technologies to have caused severe losses to the poultry industry, issues linked to AI surveillance and outbreak combat avian influenza (AI) infections. This goal its stakeholders and, ultimately, to the EU diagnosis and management, on which scientific will be achieved through the interaction of leading taxpayer. In addition, the ongoing Asian H5N1 knowledge is currently lacking. To complement European institutes along with the collaboration outbreak is a serious concern for food security this we will also develop and validate laboratory of non-EU laboratories experienced in AI outbreak and human health. It is estimated that since 2000, tests that can be used as tools in early warning control and management. more than 200 million birds have died or have systems and surveillance programmes for AI, been culled following infection with influenza in the presence and absence of vaccination. A study will be conducted to establish the viruses subtypes H5 or H7. Approximately Protocols will be harmonised and applicable to effectiveness of the current EU surveillance 50 million of these birds were from Europe. surveillance of wild birds and to different areas of and early warning systems for AI and then to Importantly, human infections have also been the poultry industry. develop blueprints for improvements to these reported in several of these outbreaks. In Asia, programmes in disease-free periods and during due to both social conditions and the particular outbreaks. The model will include criteria for characteristics of the H5N1 virus, the crossing of Expected results: harmonised diagnostic tests for on-farm outbreak the species barrier represents a serious potential We will establish comprehensive, harmonised, investigation. risk of a new human pandemic virus emerging. validated AI surveillance, ‘early warning’ and diagnosis protocols, based on an intelligent To complement this study a range of diagnostic AI is a highly contagious trans-boundary animal framework, spanning different poultry populations tools will be developed, evaluated and disease, able to spread in a susceptible population and industry sectors in the EU. This will provide an validated alongside the evaluation of a range in a short period of time. Therefore, the prompt appropriate surveillance system model for the EU, of commercially available tests. This will include identification of infected animals is crucial for with capabilities extending beyond the immediate sophisticated laboratory based methods, high control and eradication purposes. Surveillance remit of establishing optimal surveillance throughput techniques for molecular and must be targeted to appropriate areas and strategies. serological testing, penside testing and simplified species, and diagnostic tests must be appropriate tests for use in laboratories with limited resources for the setting in which they will be used, be Such a dynamic surveillance model can also be or experience. Efforts will particularly focus on properly validated and ‘fit for purpose’. utilised to provide decision-support mechanisms the validation of tests for use on clinical materials for disease control policies and enhanced analysis derived from Anseriformes, other wild bird species of novel, emerging test technologies and emerging and some selected mammalian species. threats or variables that may be encountered in

Prof. Sándor Belák AFSSA-site de Ploufragan Dr Ben Peeters Dept of Virology B P 53 Centraal Instituut Dierziekte Ulls väg 2B 22440 Ploufragan Faculty Bureau of Veterinary Coordinator: Controle Lelystad (CIDC-Lelystad) 751 89 Uppsala France Medicine Collaborators: Dr Jill Banks P.O. Box 2004 Sweden [email protected] P.O. Box 80163 Dr Dennis Senne Virology Dept Houtribweg 39 [email protected] Dr Poul Henrik Jørgensen 3508 Utrecht NVSL Veterinary Laboratories Agency Lelystad 8203 AA Dr Chris Danks Danish Institute for Food and The Netherlands US Department of Agriculture Woodham Lane The Netherlands Forsite Veterinary Research [email protected] Animal and Plant Health Addlestone [email protected] Central Science Laboratory Denmark Cepheid Europe Inspection Service Surrey KT15 3NB Dr Celia Abolnik (CSL) [email protected] Vira-Solelh Veterinary Services UK Onderstepoort Veterinary Sand Hutton Dr Artur Summerfield Maurens-Scopont National Veterinary Tel: +44 19 32 35 73 07 Institute York YO41 1LZ Institute of Virology and France Services Laboratories [email protected] South Africa UK Immunoprophylaxis [email protected] USA [email protected] [email protected] Sensemattstrasse 293 Dr GuidoVogel [email protected] Partners: Dr Martin Beer Prof. Piero Migliorato 3147 Mittelhäusern Kantonales Laboratorium Dr David Suarez Dr Ilaria Capua Friedrich-Loeffler-Institut Polysilicon TFT Group Switzerland Basel-Stadt SEPRL Laboratorio di Virologia Federal Research Institute for University of Cambridge [email protected] Kontrollstelle für Chemie- und US Department of Agriculture Istituto Zooprofilattico Animal Health Engineering Dept Advance Nanotech Ltd Biosicherheit Agricultural Research Service Sperimentale delle Venezie Boddenblick 5a UK UK Kannenfeldstrasse 2 Southeast Poultry Research Viale dell’Università 10 17493 Greifswald – Insel [email protected] gerhard.rebel@advancenanotech. Postfach Laboratory 35020 Legnaro (PD) Riems Dr Veronique Jestin com 4012 Basel USA Italy Germany Agence Française de Sécurité Dr J Arjan Stegman Switzerland [email protected] 38 [email protected] [email protected] Sanitaire des Aliments University of Utrecht [email protected] [email protected] of avian influenza across the EU and to provide provide to EU and the across influenza avian of diagnosis the standardise and harmonise to order in validation, assay of rules OIE the following by validated, and standardised internationally be will assays The populations. avian free-living and domestic in infections virus AI of diagnosis differential and detection for techniques, molecular novel these of harmonisation and application evaluation, development, the on efforts focus isto purpose The technologies. novel and detection antigen diagnostics, molecular of issues the address will FLUTEST available and/or developed in FLUTEST. between EU Member States, using different tests developed ‘fit-for-purpose’ for different conditions the optimal lay-out of surveillance programmes, will be evaluated. This will enable us to establish and for action on disclosure of the first detected case optimum distribution of resources for surveillance and use of data in the face of an outbreak and the In addition, factors such as the efficient capture clinical inspection have on these parameters. sample sizes, sampling interval and frequency of vaccination, sensitivity and specificity of the test, we will determine the influence that factors such as surveillance programme, will be established. Then flocks at the time of detection, given the current EU and detection and the average number of infected and the average time between virus introduction European Union Member States will be provided and production sector demographics across programmes. A description of the poultry population and blueprints for improvement of surveillance quality of the current EU surveillance programme the future. A report will be prepared describing the LPAI strains. Serological surveillance programmes programmes surveillance Serological strains. LPAI for isexacerbated situation the and immediately reported be not may poultry to HPAI strains of introduction that shown has Experience field’. ‘in and partners between trialled formats, portable to transferred be will developed assays the appropriate, Where manual. EU diagnostic the in specified standard the to to according tests test could have applications for DIVA surveillance DIVA surveillance for applications have could test This antigens/peptides. recombinant same the of utilisation from benefit both LFD, as typing sub- antigen an with conjunction in developed be will antibodies specific virus detecting of capable LFD An antibodies. display phage and antigens recombinant of use the involve will This isolates. virus the subtype will that developed be will (LFD) device flow lateral A competitive results. test the interpreting when required expertise the and reagents the by influenced success, with isolates AI subtyping when difficulties experience laboratories Many programmes. surveillance in use for makecost-effective to it automated and validated developed, be will eggs in antibodies AI detect to test Alaboratory advantages. many offer could yolk egg in antibodies AI of measurement the but cost-effective, not and is impractical testing of frequency high at samples blood Using disadvantage. this counter can frequency However,ahigh-test challenge. and exposure following occur to seroconversion detectable for lag time the on based stage early avery at flock infected an detect to suited primarily not are the research will support some of the unanswered unanswered the of some support will research the of outcomes the addition, In protection. consumer and welfare and health animal of levels high of attainment the to crucial are that and unexplored and lacking currently are which knowledge of areas addresses specifically FLUTEST P and point-of-care diagnosis. than 15 minutes. Thus it would be ideal for field use device), portability and rapid results, typically in less of this technology are cost (two to three euro per technology will be developed. The advantages polycrystalline silicon-thin film transistors (LTPS-TFTs) sensing mechanism employing low-temperature detection of DNA hybridisation, and a novel based on microarrays, but using label-free electrical biosensor AI An instrumentation. portable in implement to difficult and isexpensive detection optical However,the diagnosis. differential hence and detection multi-agent enable They viruses. of characterisation comprehensive and detection the for tools powerful are microarrays DNA tests. diagnostic influenza avian as applicability their for technologies innovative explore also We will employed. are strategies vaccination when implemented more rapidly. more implemented be can procedures eradication and/or control necessary all and limited be can infection of spread possible, ismade diagnosis rapid tools, such of availability the With AI. against fight the in factor fundamental astrategically presents This diagnosis. differential and surveillance for laboratories equipped well in use for assays rapid and throughput’ ‘high with supported and complemented be will tests These disease. potential of diagnosis line front validated, for provide therefore will FLUTEST globally. laboratories small and abattoirs veterinarians, by field the in used readily be can which tools capabilities, test diagnostic and surveillance care point-of- biosecure, cost-effective, provide to tests pen-side inexpensive and user-friendly simple, novel, include will Techniques programmes. management outbreak and surveillance complement to tailored tests diagnostic validated of arange of provision and development the be will FLUTEST of impact A key strategic health. human to risks potential poses and losses economic high in resulted has which a disease for particularly standards, welfare and health animal in improvements sustained EU for the within needs the addresses project The poultry. of disease epizootic of control and prevention the to regard with Commission European the within makers decision aid will generated data The influenza. avian of threat and impacts expanding globally the of management the in importance pivotal of are that questions applications otential :

FLUTEST SSPE-CT-2007-044429 Network For Early Warning Of Influenza Viruses In Migratory Birds In Europe

Acronym: NEW-FLUBIRD EC contribution: €1 855 350 Starting date: 01/02/2007 Duration: 36 months Instrument: STREP Key words: Influenza, pandemics, vaccines

Summary: routes to Europe in areas that represent a behavioural aspects of birds related to possible NEW-FLUBIRD will establish a European network relatively high risk to Europe since they harbour contacts with domestic poultry. Surveillance sites of virologists and ornithologists, data managers, migratory birds that migrate to Europe, like in will be provided with the appropriate equipment epidemiologists and modellers, in order to provide Russia, the Middle East and Africa. This will be needed to expand wild bird surveillance for AI ‘early warning and risk assessment systems’ implemented by a network of institutes involved and standardised techniques related to sampling in real time for the threat posed to animal and in water bird monitoring and research under the and laboratory testing for AI viruses including human health by avian influenza (AI) viruses lead of Wetlands International, involving Station quantitative real-time PCR and sequence analyses, from migratory birds. The network will largely Biologique Tour du Valat (Camarque, France), as well as reporting bird morbidity and mortality. build on and extend existing collaborations Wildfowl and Wetlands Trust (Slimbridge, UK), between AI virologists in Europe and international Oiseaux Migrateurs du Palearctique Occidental In addition, wider scale mortality monitoring ornithological organisations active within and (OMPO, France), CIRAD (France) and offices will be implemented on the basis of existing outside Europe including Africa, the Middle East of Wetlands International in Wageningen, the logistics in the framework of the International and Eastern Europe. Netherlands (HQ), Kiev (Ukraine), Moscow Waterbird Census (IWC), coordinated by (Russia) and Dakar (Senegal). Wetlands International. In Europe alone this Epidemiological assessments will thus cover the global scale monitoring scheme for water birds major flyways of migratory birds over Europe and In addition, the network will interconnect with results in over 25 million water birds counted the areas from which birds in Europe migrate. other networks covering similar activities in the from over 10 000 sites annually. Systems will be Furthermore, it will focus on experimental Americas and Asia though collaborative activities put in place for real time reporting of mortality infection of selected migratory bird species with of participants of the NEW-FLUBIRD network. from this network. Observations on interaction HPAI virus H5N1 and possibly other relevant HPAI The implementation of the project has a practical between wild birds and poultry in and around viruses, to determine pathogenesis and excretion general target: the urgent development of ‘early the various poultry management systems and profiles. In turn, the ornithological studies will warning and rapid response systems’ to be situations in Europe will be obtained as a construct migratory route maps and set up used by the relevant EU and other international contribution to the assessment of risks. Data systematic sampling from healthy wild migratory organisations as well as policymakers in EU managers, epidemiologists and modellers will birds thus providing insight in volume and Member States involved in combating animal and use well-established and standardised tools and timing of migration as well as key sites of those human influenza threats posed by AI viruses from techniques. Standardisation of techniques will migratory bird species that pose the highest risk migratory birds. be achieved by targeted training courses for all of transmitting HPAI viruses to poultry in Europe. participants involved in the respective areas, Finally, NEW-FLUBIRD will seek integration with Scientific and technological objectives and by providing the laboratories and sites with global early warning systems developments like of the project standardised materials and equipment. GLEWS of FAO and WHO (Global Early Warning The scientific objective of the project is to System) and GNAIS of WCS (Global 3 Network for establish a multidisciplinary network for ‘early In addition, on the basis of the currently available Avian Influenza Surveillance). warning and risk assessment’ in real time for and emerging data from the project, experimental influenza viruses in migratory birds in Europe. The infections with a recent HPAI H5N1 virus isolate project aims to do this by gaining insight into the (e.g. turkey isolate from Turkey) will be carried Problem: role of migratory birds in the possible spread of out in a high security laboratory setting (BSL The threat posed to animal and human health by AI viruses to and among poultry and mammals 3+) in at least nine wild bird species considered AI viruses from migratory birds. in Europe with a strong emphasis on the current to pose a possible risk to domestic poultry in threat posed by the HPAI virus H5N1. Europe. Clinical signs, (histo)pathology, immune histochemistry (IHC) and H5N1 virus excretion Aim: To this end the NEW-FLUBIRD network of profiles will be monitored in these species. The NEW-FLUBIRD project, initially based on influenza virologists, ornithologists, data the work carried out in the framework of the managers, epidemiologists and modellers will Collectively, the data generated by the bird FP5 NOVAFLU project and also on the FP6 be established. Specific attention will be paid to surveillance activities and by the experimental bird EDEN and the French MigrAv project networks, the selection of surveillance sites on the basis infection experiments will form the scientific basis will be extended with additional sampling sites of geographical distribution, risk estimation, the for the main objective of this project: creation 40 positioned within Europe and along migratory volume of migrating birds of different species and of real time early warning and risk assessment system as regards the role of migratory birds in of the partners in the areas of virology and/or Partners: spreading AI viruses to and among poultry and ornithology or in the areas of data management, Ward Hagemeijer Wetlands International mammals in Europe. epidemiology and modelling. In addition, the Biodiversity and Ecological relevance of the sampling sites of migratory birds Networks P.O. Box 471 in Europe and in areas from where birds migrate 6700 Wageningen Expected results: to Europe has motivated the inclusion of several of The Netherlands The NEW-FLUBIRD network will establish ‘early the partners. Furthermore, several of the partners Tel: +31 31 74 78 867 Ward.Hagemeijer@wetlands. Dr C Terregino DVM PhD warning and risk assessment systems’ in real time are involved in other projects covering the same org Laboratorio di Virologia for the threat posed to animal and human health subject in the Americas, Africa, the Caucasus Dr T C Harder PhD Istituto Zooprofilattico Friedrich-Loeffler-Institut Sperimentale delle Venezie by AI viruses from migratory birds. and Eurasia, thus allowing the NEW-FLUBIRD Federal Research Institute for Viale dell’Università 10 network to integrate global ‘early warning and Animal Health 35020 Legnaro Boddenblick 5a Italy Exploitation and dissemination of the results risk assessment systems’ as they emerge. 17493 Greifswald-Insel Riems Tel: +39 04 98 08 43 69 of these systems will be communicated on real Germany [email protected] time basis to EU organisations like DG-SANCO, Tel: +49 38 35 17 196 Dr M Gauthier-Clerc [email protected] Station Biologique de la Tour ECDC, WHO (regional office, Copenhagen) UNEP Potential applications: Prof. B Olsen du Valat and European chief veterinary officers as well as Early warning and risk assessment systems will Kalmar University Le Sambuc Section for Zoonotic Ecology 13200 Arles international organisations like OIE (Paris) and be used by policy-makers, as input for decisions and Epidemiology France FAO (Rome). In addition, ministries of agriculture and control measures for AI viruses coming from Dept of Biology and Tel: +33 49 09 72 013 Environmental Science Gauthier-Clerc@tourduvalat. and public health of Member States will also be migratory birds. 39182 Kalmar org directly informed about the threats and risks as Sweden Dr I Brown they emerge. To this end, these organisations Tel: +46 48 04 46 195 Veterinary Laboratories Agency [email protected] Woodham Lane will be part of a direct AI early warning and risk Dr P H Jorgensen DVM, New Haw assessment mailing system and the data will PhD Addlestone KT15 3NB Danish Institute for Food and Surrey be accessible by all the participants and the Veterinary Research UK organisations via the shielded part of the NEW- Hangøvej 2 Tel: +44 19 32 35 73 39 8200 Aarhus [email protected]. FLUBIRD website that will be created specifically Denmark gov.uk for this purpose. All the data generated by the Tel: +45 72 34 68 25 Dr F Monicat project will be published in the peer reviewed [email protected] UR Gestion Intégrée de la Dr Zenon Minta DVM PhD Faune international scientific literature, after securing National Veterinary Research Montpellier intellectual property rights as appropriate. In Institute France Al Partyzantow 57 Tel: +33 14 40 10 510 addition, the public part of the website will 24100 Pulawy [email protected] inform people about the outcome of those parts Poland Dr B Hughes Tel. +48 81 88 63 051 Wildfowl and Wetland Trust of NEW-FLUBIRD that, according to the project’s [email protected] Slimbridge GL2 7BT intellectual property management rules, can be Dr Christine Monceyron Glos made public. Jonassen UK National Veterinary Institute Tel: +44 14 53 89 11 72 Section for Virology and [email protected] The added value of carrying out the work at the Coordinator: Serology Dr Alexandre Czajkowski Prof. Dr A D M E Osterhaus P.O. Box 8156 OMPO European level is illustrated by the participation Erasmus MC 0033 Oslo 5 Ave des Chasseurs SSPE-CT-2007-44490 of scientific groups from nine EU Member States 3000 Rotterdam Norway 75017 Paris The Netherlands Tel: +47 23 21 64 09 France and motivated by the specific expertise of each Tel: +31 10 40 88 066 christine.monceyron- Tel: +33 14 40 10 510 [email protected] [email protected] [email protected] NEW-FLUBIRD Avian Influenza Virus Survival In Poultry Commodities, Poultry Manure And The Environment

Acronym: FLURESIST Key words: Avian influenza, poultry commodities, EC contribution: €870 000 Starting date: 01/03/2007 virus survival, bio-security measures, disinfection, Duration: 36 months Instrument: STREP cleansing operations, public health risk

Summary: Problem: Aim: Avian influenza (AI) outbreaks have recently The circulation of the HPAI virus in Asia and now The aim of the project is to obtain data and caused severe losses to the poultry industry, its also in the Middle East and Africa could represent provide knowledge about the presence of stakeholders and, ultimately, to the EU taxpayer. the origin of a pandemic virus for humans, and a influenza viruses in commodities and litter of In addition, the ongoing Asian H5N1 outbreak great number of questions have been raised with infected poultry. In order to develop validated is a serious concern for food security and human a view to finding a way to combat the ongoing AI protocols for cleansing, disinfection and treatment health worldwide. crisis. Due to the lack of field and experimental of litter and to be able to assess the risk of carcass data certain questions on virus survival in the disposal, treated litter, and poultry commodities In Asia, due to both social conditions and the environment and in poultry and other avian such as meat, feathers and eggs, the virus survival particular characteristics of the H5N1 virus, the commodities are not yet answered and these will be determined in a standardised manner in crossing of the species barrier represents a serious knowledge gaps should be filled following the different environments. The project also aims to potential risk of a new human pandemic virus results of the ongoing and new research efforts of create knowledge about environmental factors emerging. Evidence is growing that HPAI H5N1 is the scientific community. that influence virus stability. Data collected will not only spreading by trade but is also carried by be used for proper risk assessment of the trade wild birds. H5N1-infected wild birds, mainly water in treated and fresh poultry commodities, poultry fowl, have recently been detected in the European litter and the contaminated environment. Union in Italy, Austria, Germany, France, Greece, Sweden and Poland. These findings are raising our awareness that H5N1 is becoming more and more endemic in wild birds. The finding of a cat, stone marten and raptors that died as result of infection with H5N1 has uncovered the consequences of this development.

More questions are being raised about the risk of contamination of surface water in relation to the health of other animals and humans. To answer these questions and to be able to assess the risks involved in trading in poultry commodities and litter, more knowledge about virus content of commodities, the stability of the virus in these products, in litter and the environment is needed.

42 assessments. risk make to proper used be will Data validated. or and adapted be will disinfection and disposal carcass treatment, waste for protocols results the on Based survival. virus decreases or increases pollution sewage and flora gut or faeces sterile waterfowl, of factors enteric organisms, living and lake silts soils, certain whether on obtained be will knowledge Furthermore, products. poultry treat to used temperatures at and temperatures ambient at commodities these in viruses of survival on data quantitative generate will Studies determined. be will eggs and feathers meat, as such commodities poultry in concentrations Virus survival. on temperature and pH as such parameters physical of effect the and viruses, influenza avian different of survival on data provide will Research Ex results pected : measures of bio-security. Any such response must must response such Any bio-security. of measures further of development the for abasis and response any of formulation the underpin to data scientific the provide to isdesigned project this and problems of anumber faces society threats, these of view In man. to eventually perhaps and industry, poultry the to threats serious poses influenza avian from threat current The P and products, and thus help calm public concerns. concerns. public calm help thus and products, and materials avian particularly materials, different in virus of stability and levels the on data provide to isdesigned project this reasons these For soil. in survival virus of possibilities even and water surface and ground for problems contamination potential avoid to are we if considered be to have this of consequences virological the and litter and carcasses of quantity alarge of dispose to a need be would there outbreak an of event Finally, the in home. at consumption for even addressed be to have poultry, vaccinated of eggs or meat even and poultry, of safety the over misgivings consumer Similarly, industry. the in engaged those for consequences severe have and capacity trading anation’s restrict certainly would outbreak an industry: the of competitiveness the threatens turn in this trade; or populations wild from virus of introduction the include industry to Threats both. with interact humans which in way the to relation in particularly and environment the and host its in both virus the of behaviour the of understanding an by driven be applications otential : Houtribweg 39 Houtribweg 2004 Box P.O. (CIDC Lelystad) Lelystad Dierziekte Controle Instituut Centraal Ph Koch Guus Coordinator: Tel: +44 14 83 68 97 19 97 [email protected] 68 83 14 +44 Tel: UK Surrey 7XH GU2 Guildford Hill Stag Sciences and Molecular Biomedical of School Surrey of University Ph JCarter Michael 19 [email protected] 43 67 18 +46 Tel: Sweden Uppsala 89 751 2B vag Ulls (SVA) Anstalt Veterinarmedicinska Statens O Jakob [email protected] 602 [email protected] 81 96 23 +97 Tel: Israel Dagan Bet 50250 Box Diseases P.O. Avian of Division Institute (KVI) Veterinary Kimron I 39 73 [email protected] 32 19 +44 Tel: UK 3NB KT15 Surrey Addlestone Lane Woodham Dept Agency Virology Laboratories Veterinary I 69 43 [email protected] 08 98 04 +39 Tel: Italy (PD) Legnaro 35030 Virologia di Venezie Laboratorio delle Sperimentale Zooprofilattico Istituto I Partners: 800 [email protected] 38 02 32 +31 Tel: Netherlands. The Lelystad AA 8203 laria Capua Capua laria an Brown Ph Brown an rit D rit avidson Ph avidson ttoson Ph ttoson D D D D D t I [email protected] Canada 8P9 K2H 3851 Ottawa Road Fallowfield (CFIA) Agency Inspection Food Canadian S Loyd 433 [email protected] 63 54 06 +17 Tel: USA 30605 GA Road Athens Station College 934 (USDA-ARS/SPRL) Research Laboratory Poultry Service Southeast Research Agriculture of Agricultural Department US D ES n c h e p o wayne Ph wayne ro ll pencer Ph pencer a j e b c oration w oration t: D D it h FLURESIST SSPE-CT-2007-044311 www.fluresist.eu Resistance of Influenza Viruses in Environmental Reservoirs and Systems

Acronym: RIVERS Key words: Influenza, avian, pathogenic, zoonotic, EC contribution: €1 395 000 Starting date: 01/02/2007 resistance, inactivation, virus, environment, Duration: 36 months Instrument: STREP reservoirs, systems, models, restocking

Summary: evidence-based guidelines for the prevention c) Data on the prevalence of AIVs in waters The surge of the global avian influenza and control of influenza outbreaks in animal (lakes, ponds and rivers) in the course of time epizootic mainly caused by the genotype Z and human populations, especially at times of throughout the year and along the stream of high pathogenic avian influenza virus (HPAIV) restocking. rivers; data on gastropods and bivalve molluscs has posed numerous questions, in particular regarding their potential role as concentrators to risk managers and policymakers. Scientific and reservoirs of AIVs in aquatic biotopes. knowledge is thin on many aspects of the Problem: d) Data (database) on IV survival in the air and ecology and environmental properties of Highly pathogenic avian influenza epizootics on various kind of surfaces and in various HPAIVs, in particular H5N1. Virus survival, a key associated with zoonotic human cases. conditions, data on the prevalence of AIVs element in control strategies, is an illustration in the surroundings of farms with present of this paucity of knowledge. Data from the and past outbreaks in the course of time literature on AIV survival is rather limited, often Aim: throughout the year. very old and sometimes not confirmed from one The overall aim of this project is the prevention e) Descriptive, data driven, low-level simulation study to another or even contradictory. and control of avian influenza A(H5N1) in the models of AIVs perpetuation, viability and animal population through the following specific deactivation in (i) various water environments; The results obtained with various sub-types objectives: laboratory-controlled and natural; (ii) in air of influenza A viruses cannot be extrapolated a) Gathering data on the survival of at laboratory-controlled environments; (iii) in to the current A (H5N1) viruses before careful avian influenza viruses (AIV), in natural avian faeces and farm manure. consideration. Furthermore, little information is environments. f) Multi-scale agent-based simulation model provided regarding the survival of IVs in the air b) Generating scientific knowledge about of possible determinants for AIV’s stability, and surfaces - no standardised protocols exist the survival of avian influenza viruses in perpetuation and deactivation. to detect AIVs in waters, in the air or in/on solid experimental settings. matrices. Ideally, the virus detection technique c) Providing figures about the effect of various to be used should be sensitive, quantitative, treatments either chemical (e.g. disinfectants) Potential applications: rapid and applicable in routine before or after a or physical (e.g. UV light) on influenza virus a) Recommendations for the prevention and standardised sampling method, including or not survival. control of current and future avian influenza concentration. d) Providing figures on the effect of various types outbreaks in wild and domestic birds with a of food processing on influenza virus survival. pandemic potential in Europe and the rest of Under this project, nine institutions directly e) Elaborating models about the survival of AIV the world will be drawn from the data obtained involved in AIV, three from Asian countries, in natural environments to demonstrate, in through RIVERS in a final report to the EC to have joined forces in order to investigate the connection with other EU projects in this area, allow evidence based policymaking. prevention and control of influenza outbreaks in their perpetuation in nature both in biological b) International guidelines for the control and the animal population at present and at time of and environmental reservoirs. prevention (through virus inactivation and restocking. More specific objectives are: disinfection for example) of outbreaks in domestic birds but also in humans will benefit a) to understand the basis of virus survival from Expected results: from the data generated by the project. a virological viewpoint; a) Criteria for bio-equivalence in relation to virus b) to understand the impact of physical and survival between IV strains. chemical elements on virus survival; b) Method of virus viability assessment other c) to evaluate the role of environmental than virus titration on cell culture, approvable reservoirs; and standardised protocols for influenza virus d) to propose standardised protocols for recovery from various surfaces, approvable the concentration and detection of AIVs and standardised protocols for testing the in waters, including waste waters, and in effect of chemical and physical treatments different matrices including food; of different types of water on influenza virus e) to provide a database together with survival, Standard Operating Procedures for virus 44 analytical tools to allow the generation of disinfection/inactivation in different settlements. [email protected] www.pasteur-kh.org Cambodia Penh Phnom Boulevard Monivong 5 983 Box P.O. IPC Cambodge I [email protected] www.microbio.bas.bg Bulgaria 26 Sofia Str 1113 Bonchev G Acad SAIM T [email protected] www.cantacuzino.ro/en/ Romania Bucharest 050096 5 District 103 Independentei Splaiul 1-525 Box P.O. Immunology Microbiology and Research Development of and Institute National Cantacuzino Partners: 807 [email protected] 13 06 14 +33 Tel: France Paris Roux 75724 Docteur du Rue 25 Pasteur Institut Manuguerra Jean-Claude Coordinator: I nstitut Pasteur du du Pasteur nstitut nstitute of Microbiology of nstitute he S he tephan A tephan ngeloff ngeloff [email protected] www.icm.edu.pl Poland Warsaw 02106 Bldg 5a Pawinskiego ICM Modelling and Computational Mathematical Centre for Interdisciplinary Warsaw of University [email protected] France Lille 59019 Calmette Rue 1 245 Box P.O. Lille de I [email protected] www.cirad.fr France Montpellier 34398 30/E TA de Baillarguet International de Campus CIRAD R I Coopération de Centre [email protected] [email protected] China Road Shanghai Chaongging South 225 Shanghai of IPS-CAS Institute Pasteur S A Chinese nstitut Pasteur Pasteur nstitut en nternationale ciences echerche A echerche cademy of of cademy gronomique [email protected] www.whiov.ac.cn China Wuhan 430071 44 Virology of Xiachongsam Institute Wuhan S A Chinese ciences cademy of of cademy

RIVERS SSPE-CT-2006-044405 www.rivers-project.eu New and emerging technologies: improved laboratory and on- site detection of OIE list A viruses in animals and animal products Acronym: LAB-ON-SITE Key words: Diseases notifiable to OIE, EC contribution: €1 500 000 Starting date: 01/11/2004 transboundary animal diseases, viruses, diagnosis, Duration: 39 months Instrument: STREP diagnostic tests, dipstick, emerging technologies

Summary: Expected results: In order to produce an internal control for the AIV This project is improving the diagnosis of nine a) Improved laboratory methods for sensitive PCR, a ‘mimic’ has been constructed, which is major transboundary animal diseases (TADs) — diagnosis of nine TADs notifiable to OIE. assuring the reliability of the diagnosis. foot-and-mouth disease, swine vesicular disease, b) Improved, commercially available dipstick tests Rapid one-step real-time PCR (RT-PCR) for the vesicular stomatitis, classical swine fever, African for on site diagnosis. detection (pathotyping and sub-typing) of HPAIV swine fever, bluetongue, African horse sickness, c) Methods to analyse animal products for the using SYBR Green and TaqMan chemistries is Newcastle disease and highly pathogenic avian presence of the targeted viruses. under development. influenza — by addressing the recommendations d) Sample banks, standardisation, validation, of the Scientific Committee on Animal Health dissemination of results, SOPs, international 2. Development of rapid, simplified and Animal Welfare (SCAHAW). Regarding its training. method for the on-site detection of epidemiological importance, swine influenza is highly pathogenic AIV (HPAIV) antigens also included in the project. The programme is Work performed on avian (dipstick tests) focusing on new and emerging technologies, influenza viruses (AIV): specifically the development, validation and Two monoclonal antibodies (MAbs 5F10, and dissemination of robust, specific and sensitive 1. Rapid detection of AIV with light HB-65) were evaluated in dipstick tests. Devices diagnostic tests. upon extension primers and a duplex for lateral chromatography were prepared using RT-PCR either of the MAbs and specificity testing was The project is combining the development of carried out. The obtained results show that only improved ‘front line’ diagnostics, such as dipstick A real-time RT-PCR assay utilising light upon MAb 5F10 gave a positive result using the HPAIV tests that can be used by veterinarians in the extension fluorogenic primers (LUX RT-PCR) was strains while MAb HB-65 gave failed to react. field with development of robust and simple developed for the rapid and efficient detection MAb 5F10 was able to detect all three HPAIV nucleic acid and antigen-antibody detection of AIV (Kiss et al., 2006). The LUX RT-PCR strains and the negative controls gave a negative methodologies to be applied in local laboratories technology provided a highly specific and result in the experiment. and in abattoirs. In addition, the most recent sensitive novel means of AIV detection. Besides approaches in real-time PCR and in microarray its simplicity (use of only two oligos), the LUX 3. Development of ELISA assays for technologies are applied for the improved technology efficiently reduces the likelihood of HPAIV typing and antigen detection diagnosis of the nine TADs, notifiable to OIE. false positive results by the use of melting point analysis upon amplification. The objective is the development of ELISAs to Problem: identify H5 and H7 viruses using monoclonal TADs are regularly emerging and re-emerging, A duplex reverse transcription-polymerase chain antibodies specific for the H5 and H7 threatening animal and human health worldwide reaction (dRT-PCR) assay has been developed for haemagglutinin antigens, respectively. Two MAbs (www.oie.int/). To combat these diseases more the simultaneous, rapid and specific detection of out of 42 generated hybridomas specific to H7 effectively, the development of rapid, highly AIV and Newcastle disease virus (NDV) in clinical and 20 specific to H5 were selected. Sandwich sensitive and specific novel diagnostic methods is specimens (Farkas et al., in press). The assay is ELISAs were designed for H5 and H7 antigen necessary, including simple and powerful on site robust in a sense that it is capable of detecting a typing, using a unique MAb, coated to microplate tests. broad range of AIV and NDV variants in the same wells as antigen capture antibody, and conjugated reaction. The speed, simplicity and complexity of with peroxidase as tracer. MAbs 5D8 (H5-specific) Aim: the assay facilitate the immediate and front line and 7A4 (H7-specific) were used. The aim is the complex diagnosis of nine TADs diagnosis of AI and ND close to outbreak cases. notifiable to OIE. The project is focusing on the Analytical sensitivity. Allantoic fluids development, validation and dissemination of The assay was shown to be capable of detecting containing each of four H5 and three H7 subtypes robust, specific and sensitive diagnostic tests, either virus in faecal samples. That makes it an were examined. The detection limit of H5- and including ‘front line’ diagnostics, such as dipstick appealing tool among the diagnostic procedures H7-ELISA corresponded respectively to 104,5/105 tests. used in monitoring programmes. Therefore, the EID50/0,1ml and 104/104.5 EID50/0,1ml. described dRT-PCR provides a useful and practical supportive method for the effective diagnosis of Analytical specificity. To evaluate the 46 avian influenza and Newcastle disease. analytical specificity the number of AIVs standardisation and diagnostic application was was application diagnostic and standardisation of terms in assays of Improvement instruments. sophisticated more with equipped not laboratories in also and procedures test diagnostic line first as used be can which tests, simple robust, rapid, represent they At present, project. the of year second and first the during improved and developed were above H7H5 described and serotypes on focus main with AIV of antibodies and/or antigens of detection the for assays ELISA high. very are test HI to respect with evaluated ELISA the of performances specificity and sensitivity that so significant, strongly was H7 H5 to AIVs and specific antibodies of detection the for test HI the and ELISA competitive the between correlation the conclusion, In H7. than different subtypes of AIVs to positive 176 strongly sera including H7-ELISA, in also H7-HIin negative were negative sera 396 all specificity, Concerning test. HI in titre athreshold showed turkeys vaccinated from samples sera Two missed H7-ELISA. by 417of detected H7-HI in not were positive sera 2out whilst H5-ELISA in also positive scored H5-HI in 173 All positive sera tested. were AIVs of subtypes heterologous with infected animals from originating sera and (n=220) samples naive both analyses, specificity For sera. positive experimental and natural both on test reference as test (HI) inhibition haemagglutination using evaluated were ELISAs two the of specificity and ELISAs competitive H7 and H5 Hsubtype. respective the with tested strains all on present epitope an recognise and activity HI have ELISAs antibody-detection for selected MAbs 7A4 and (H7) (H5) 5D8 The antigens. as strains AI of H7 H5 subtypes and the using assays HI by tested were 813 samples serum Total well. as developed were viruses influenza H7 H5 to and elicited antibodies of detection the ELISAs antibody-based Monoclonal infection. acute during specimens clinical in and eggs embryonated chicken SPF in isolated viruses identify to suited are ELISAs H7 typing and H5 Then, ELISAs. detection antigen for expected as signal, apositive produce to isneeded antigens of concentration asignificant sensitivity, analytical Concerning examined. H7 H5 AIVs and the all of identification the enabled that reactivity subtype intra- abroad with combined selected, MAbs two the by conferred specificity, high avery showed developed H7 H5 ELISAs and the conclusion, In Hantigens. other displaying subtypes with contrast in H7N7, H7-ELISA, the in positive were 1/56 Praga strain equine the including subtypes, H7- all Similarly, negative. reacted that AIV other any with contrast in H5-ELISA, the in reactivity strong exhibited H5 subtypes the All H7 ELISAs. H5 and both in tested was above mentioned . Sensitivity . Sensitivity for for One of the most important issues in the control control the in issues important most the of One EU. the outside and in both authorities health animal and institutes partner to disseminated be will methods diagnostic new developed The P available. now are antibodies monoclonal specific and performed Vizcaíno D Vizcaíno MS José Prof. 93 [email protected] 79 34 72 +45 Tel: Denmark Kalvehave 4771 Lindholm Virology of Dept Research and Food for Veterinary Institute Danish Ph Uttenthal Åse 79 56 [email protected] 52 90 28 +44 Tel: UK Ireland Northern 3SD BT4 Belfast Road Stoney of Belfast University Queen’s Science The Veterinary of Dept A Gordon Partners: 35 [email protected], 41 67 18 +46 Tel: Sweden Uppsala 75189 2B väg Ulls Virology of Dept Sciences Agricultural of University Swedish The & Institute Veterinary National The Ph S Prof. Coordinator: diagnosticians. line’ ‘first the for provided be will methods effective and Simple methods. such use easily can abattoirs and laboratories equipped simply veterinarians, Field project. this in developed be to instruments PCR real-time portable on based tests diagnostic or tests dipstick like the tests inexpensive and use to easy simple, very of availability EU isthe the outside countries in diseases OIE-listed of Tel: +46 18 65 49 00 [email protected] 49 65 18 +46 Tel: Sweden 32A Uppsala väg 75183 Hammarskjölds AB Dag Biotech Svanova A 33 [email protected] 81 41 52 +36 Tel: 2 Hungary Debrecen 4002 51 Pf of Debrecen Institute Veterinary I 082 [email protected] 44 39 91 +34 Tel: Spain Madrid s/n 28040 Hierro de Puerta Avda Animal Sanidad Dpto Veterinaria de Facultad de Madrid Complutense Universidad stván Kiss D Kiss stván nn N nn applications otential D , DS ordengrahn Ph ordengrahn ándor Belák D Belák ándor c llan B llan VM, Ph VM, VM, Ph VM, ánchez- S c, Ph c, D D D VM, VM, D D Italy Brescia 25124 7/9 Bianchi v/a Romagna Lombardia dell’Emilia e della Sperimentale Zooprofilattico Istituto E [email protected] 28 84 [email protected] 18 37 45 +49 Tel: Germany Borstel 23845 22 (RCB) Borstel Parkallee Center Research Prof. Jabbar Subc 910 [email protected] 14 47 18 +46 Tel: Sweden Uppsala AB 75185 Bioscience Olink R&D of Head Ph M Landegren Ulf Prof. Subc 53 60 [email protected] 02 60 12 +43 Tel: Austria Vienna 1400 100 Box 5 P.O. Wagramerstrasse Division IAEA Joint FAO/IAEA and Section Health Production Animal of Head Prof. Gerrit Viljoen Ph Th 31 11 [email protected] 23 83 14 +44 Tel: UK Surrey ONF GU2 Pirbright Road Ash Laboratory Health Pirbright Animal for Institute D 66 [email protected] 73 64 92 32 + Tel: Belgium Merelbeke 9820 133 Medicine Salisburylaan Veterinary of Faculty Virology of Laboratory University Ghent D R Van Kristien Prof. 310 [email protected] 90 22 30 +39 Tel: Ph miliana Brocchi Ph Brocchi miliana VM, Ph VM, onald King Ph King onald D D ird p ird ontra ontra arty: arty: D A c c hmed tor 2: tor 1: tor D : eeth eeth D D VM, VM, D D , DSc ,

LAB-ON-SITE SSPE-CT-2004-513645 www.labonsite.com PATHOGENESIS AND IMPROVED DIAGNOSIS AND CONTROL OF AVIAN INFLUENZA INFECTIONS

Acronym: AVIFLU EC contribution: €1 839 294 Starting date: 01/10/2002 Key words: Avian influenza, poultry, highly Duration: 48 months Instrument: Shared Cost Action pathogenic, pathogenicity

Summary: Problem: Aim: In recent years there has been a global increase Control of an infectious disease that can spread as a) To develop rapid and sensitive laboratory tests in the number of outbreaks of highly pathogenic rapidly as HPAI requires a rapid and harmonised for avian influenza (AI) for use in surveillance avian influenza (HPAI). Two devastating response. It is critical that veterinarians and and controls programmes. outbreaks have occurred in Europe - in Italy animal keepers recognise the clinical signs of the b) To develop a transmission model to elucidate and the Netherlands - both resulting in major the disease and report suspected cases quickly. the dynamics of influenza A virus infection in economic losses. In Asia an ongoing epizootic Suspected outbreaks must be backed up by rapid, chickens and waterfowl and to use this model is causing major concern due to the number of sensitive and highly specific laboratory tests to examine the emergence of highly pathogenic human infections associated with a particular before control measures can be implemented. viruses from those of low pathogenicity, strain of H5N1 AI virus, sparking fears and and to measure the efficacy of vaccines and considerable planning for a potential influenza Early warning systems such as surveillance in wild vaccination strategies. pandemic. Millions of birds have been culled to bird reservoirs and in poultry flocks also play an c) To develop and evaluate the use of AI marker control these outbreaks. important part in the control of AI. This requires vaccines. the testing of high volumes of samples in a d) To study the pathogenesis of AI in animal Much attention has focused on determining the timely manner and may require a different testing models and the molecular basis of virulence. best control strategies for dealing with outbreaks strategy to that of index cases. It is also crucial e) To study AI pathogenicity, transmission and and new introductions of HPAI viruses in poultry. that we understand the mechanisms and markers the efficacy of vaccination in ducks. Successful control strategies depend upon a for the emergence of HPAI and the impact number of factors including the use of rapid and vaccination can have on the spread of disease. sensitive laboratory tests appropriately applied; knowledge about the infecting virus, e.g. the host species affected; rates and mechanisms of transmission between animals of the same Coordinator: Dr Jill Banks or different species; and determinants for Veterinary Laboratories Agency pathogenicity and the use of vaccination. Dept of Environment, Food and Rural Affairs, Virology Dept Woodham Lane We have addressed some of these issues in the Addlestone Surrey KT15 3NB AVIFLU project: Rapid and sensitive laboratory UK tests for avian influenza (AI) for use in surveillance Tel: +44 19 32 35 73 07 and controls programmes were developed and [email protected] validated. A transmission model was used to Partners: study the efficacy of vaccination in poultry and Dr Poul H Jorgensen Danish Veterinary Laboratory ducks, AI marker vaccines were developed and Dept of Poultry Diseases evaluated, and reverse genetics was used to study Hangoevej 2 Aarhus N the effect of molecular pathogenicity markers in Dk8200 Denmark vitro and in vivo. [email protected] Dr Guus Koch CIDC-Lelystad Dr Ilaria Capua 2004 Istituto Zooprofilattico Houtribweg 39 Sperimentale delle Venezie 8203 AA Lelystad Virology Dept The Netherlands Via Romea 14/A [email protected] 35020 Legnaro Dr Véronique Jestin Italy AFSSA Laboratoire D’Etudes [email protected] et de Recherches Avicoles et Hans-Dieter Klenk Porcines Philips-Universitaet Marburg BP53 Institut fur Virologie Rues des Fusilles Robert-Koch-St 17 22440 Ploufragan 35037 Marburg France Germany 48 [email protected] [email protected] Expected results: d) H7 HPAI infection of golden pheasants caused our testing regimes up to date with advances in The AVIFLU project has now been completed mortality and vaccination protected birds molecular diagnostics. and some elements of the work are now being from clinical signs but spread of the virus still explored further in the follow-on project, FLUAID. occurred. Monoclonal antibodies produced in this project Highlights of our findings are summarised below: e) Following infection of Pekin ducks with HPAI have been evaluated for their potential as A/duck/Vietnam/12/2005 H5N1 in naïve birds, reagents in a penside antigen detection test, in Diagnostic tests clinical signs, with a predominance of nervous the form of a Lateral Flow Device. The test is now Two ring trials were conducted that involved six signs, were observed starting on day two being developed further and validated within the EU national AI laboratories and a number of other post infection. The overall mortality rate was current FLUAID project. It is likely that this test invited laboratories, with the aim of establishing 70%, indicating that Pekin ducks are clinically will be marketed commercially in the future. an agreed approach to AI diagnostic RT/PCR susceptible and may die following infection in the EU. The trials were designed to evaluate with certain strains of HPAI H5N1. In situ Differences in the transmission of highly RT/PCR and real time RT/PCR protocols for the hybridisation showed a strong neurotropism for pathogenic avian influenza (HPAI) and low detection of H5 and H7 AI viruses that had been this virus in ducks. pathogenic avian influenza (LPAI) strains developed by each participating laboratory. The f) Vaccination of Pekin ducks with a single may play a role in the mechanism of selection best performing protocols for sensitivity and heterologous dose of commercial inactivated by which HPAI viruses arise in the field. The specificity were then validated and circulated to H5N2 vaccine and challenged with HPAI H5N1 mathematical infection model will contribute to all EU AI national laboratories. They are now (Asian lineage) vaccination prevented death an understanding of these processes. It is a tool included in the EU diagnostic manual which and reduced the shedding of virus but not that enables the effect of different intervention supports the new AI directive. sufficiently to prevent spread of the infection strategies to be measured, e.g. vaccination. to other animals. In contrast, a two-dose A generic AI antigen capture ELISA had been vaccination programme commencing at one Our results indicate that vaccination of Pekin developed using as a key reagent AI Nuclear day old, prevented shedding, clinical signs ducks with an inactivated, conventional product Protein (NP) specific monoclonal antibodies and mortality. This evidence suggests that the is successful in preventing clinical signs and (Mabs) developed within the AVIFLU project. perpetuation of the infectious cycle may be mortality and in suppressing shedding of viable These NP Mabs have been evaluated for their interrupted by optimum vaccination. virus. This suggests that the perpetuation of the potential as reagents in a penside antigen infectious cycle may be interrupted by optimum detection test, in the form of a Lateral Flow Molecular basis of pathogenicity vaccination. Device. Prototype devices were produced, tested A reverse genetics system was established based and compared with other devices available on viruses from the 1999-2000 Italian H7N1 These findings have significant implications on commercially. The prototypes performed well in LPAI and HPAI outbreaks. This was used to practical aspects of AI control. It appears that comparison with other devices for cloacal and investigate the biological relevance of various following field challenge a vaccinated flock will oropharyngeal swabs collected from chickens and mutations observed between these viruses. The not shed enough virus to infect either vaccinated Pekin ducks. findings were that additional glycosylation of or unvaccinated flock-mates. As the amount of the haemagglutinin near the receptor binding virus shed in the environment is not sufficient to Vaccination site promotes the spread of infection by HPAI infect unvaccinated ducks, it could be speculated An infection model was developed enabling the virus, apparently due to increased virus release that such a practice would presumably reduce quantification of different intervention strategies from cells and not due to changes in receptor the risk of infection spilling over into the wild bird QLK2-CT-2002-01454 on the transmission of avian influenza viruses. specificity. Also, a shortened neuraminidase population, and possibly to mammalian hosts. A stochastic susceptible, latently infected, stalk is a prerequisite for adaptation to chickens. infectious, recovered (SEIR) epidemic model was Viruses with a reconstituted full NA stalk had a The vaccination protocol under study was also developed and used to analyse data generated loss of infectivity and grew to lower titres. developed in order to evaluate a system that with the infection model. The transmission could be compatible with husbandry practices. experiments offer a way to measure the spread Mutations found in the NS1 proteins of high Vaccination at one and 30 days old appears to be of virus under experimental conditions and also and low pathogenic viruses seem to be a critical compatible with Pekin duck husbandry methods in to quantify the effect of vaccination on the determinant for host adaptation. Asia. The results indicate that such a programme transmission characteristics of the viruses. The could be used in the field to prevent primary model was used to investigate the transmission introduction and secondary spread in naïve Pekin characteristics of LPAI and HPAI viruses of the Potential applications: ducks. A duration of immunity above threshold H7 and H5 subtypes in vaccinated and non All the diagnostic tests and reagents (including levels for more than five months was recorded, vaccinated chickens, Pekin ducks, teal and golden monoclonal antibodies) developed in this project which appears to be longer than the economic life pheasants. are now recommended for use in the national of a meat duck, approximately four months. avian influenza reference laboratories of the EU. Vaccination may also prevent the establishment of The major findings are: The protocols are available in the EU diagnostic viraemia and therefore prevent viral colonisation a) Vaccination with heterologous vaccines manual. This effort has harmonised the approach of internal organs, positively influencing the food - suitable for use in a DIVA (differentiation taken by EU national AI laboratories and enabled security of duck products. of infected from vaccinated animal) strategy the best molecular tests currently available to be protected chickens against disease and used throughout the EU. The experience gained The reverse genetics system has already given mortality but also reduced the spread of virus in running the ring-trials is now used by the AI some surprising insights into the determinants within a group of birds. Community Reference Laboratory to conduct for host range and pathogenicity. Some of b) In ducks HPAI H5 and H7 viruses were more the EU national AI laboratory annual proficiency the markers identified may be relevant for the often shed via the respiratory route. panel testing for AI molecular tests. The strategy development of new diagnostic tests and for the c) H7 HPAI infection of teals did not cause clinical signs adopted for these trials will also ensure that surveillance of avian influenza. and vaccination prevented spread of the virus. improved tests will be identified, thus keeping AVIFLU European surveillance network for influenza in pigs

Acronym: ESNIP Key words: Influenza, swine, surveillance, EC contribution: €269 984 Starting date: 01/01/1999 epidemiology, evolution, diagnosis, public Duration: 36 months Instrument: Concerted Action health risk

Summary: Problem: Aim: A network of laboratories working in the field of Swine influenza viruses (SIVs) are enzootic in The main objective of this concerted action was swine influenza was established and harmonised swine dense regions of Europe and they are a to standardise protocols used for the diagnosis swine influenza diagnostic tests. Using these tests major cause of respiratory disease in fattening and characterisation of SIVs used in different a preliminary surveillance of influenza in pigs was pigs. Until recently, however, there was no laboratories and to define standard reagents and performed, and swine influenza viruses isolated organised surveillance for influenza viruses of make these available to all participants. Other in the last decennia were further characterised swine, as is the case for human and equine aims were: antigenically and genetically. influenza viruses. In addition, there was no standardisation of diagnostic techniques or of a) finalised requirements for antigenic the techniques used for antigenic and genetic characterisation; characterisation of swine influenza virus (SIV) b) finalised requirements for genetic strains. Because of this lack of organisation and characterisation; harmonisation of SIV surveillance, it was difficult c) database and data registration; to make recommendations for the control of SIV in d) evaluation and SI epidemiology, and Europe, and for the selection of vaccine strains in recommendations for SI control. particular. These needs have led to the submission of a proposal for an EC concerted action by researchers from several European countries.

50 nl . wur . esnip . www

Coordinator: Results: Dr G Koch The objective has been achieved. Selected CIDC-Lelystad Postbox 2004 SIV isolates from different countries were 8203 AA Lelystad compared antigenically and genetically. The use The Netherlands Tel: +31 32 02 38 609 of standardised protocols and reagents allows [email protected] the comparison of SI viruses isolated during the project period and in the future. Partners: Dr Ian Brown Veterinary Laboratories Agency Dr A Bøtner One of the problems was that only a low number Woodham Lane Danish Veterinary Institute for New Haw Virus Research of SIV isolates was available from some countries, Addlestone Lindholm 4771 because of few diagnostic submissions for SIV. Surrey KT15 3NB Kalvehave UK Denmark [email protected] Tel: +45 55 86 02 00 A comparative serosurveillance study for SIVs in Dr Kristien Van Reeth [email protected] different countries was performed, but not all Ghent University Dr Z Pospisil Faculty of Veterinary Medicine Institute of Infectious Disease partner countries were able to participate. To get Laboratory of Virology and Veterinary Epidemiology a more complete picture of the SIV epidemiology Salisburylaan 133 Palackeho 1-3 B-9820 Merelbeke 612 42 Brno in the major swine producing countries of Europe, Belgium Czech Republic researchers from Spain (Laboratorios Hipra, [email protected] Tel: +42 54 15 62 305 Dr G Barigazzi and [email protected] Girona) and Germany (Impfstoffwerke Dessau, Emanuela Foni Dr I Markowska-Daniel Rodleben), who were not official partners of Istituto Zooprofilattico National Veterinary Research the ESNIP project, were also involved in the Sperimentale della Lombardia Institute e dell’Emilia Romagna Partyzantow 57 serosurvey and/or collection of SIV isolates. This Sezione Diagnostica di Parma 24-100 Pulawy allowed us to get a more complete picture of the Via dei Mercati Poland 13/A Tel: +48 81 88 63 051 ext 232 SIV epidemiology in Europe. 43100 Parma [email protected] Italy Dr V Ohlinger [email protected] BioScreen European Veterinary A website (www.esnip.wur.nl ) is hosted by one Dr François Madec Disease Management Center QLK2-CT-2000-01636 of the partners. All protocols and a summary of Agence Française de Sécurité Gmbh the SI isolates made in the period 2000-2003 Sanitaire des Aliments Medelstraße 11 Laboratoire d’Etudes et de 48149 Muenster is available to the public on this website. A Recherches Avicoles et Porcines Germany more detailed electronic database is password Unité d’Epidémiologie et Bien- Tel: +49 25 19 80 19 00 Etre Porcin [email protected] protected and accessible only to the partners. Zoopôle Les Croix Dr F Madec BP 53 Agence Française de Securité 22440 Ploufragan Sanitaire des Aliments France Laboratoire d’Etudes et Potential applications: [email protected] de Recherches Avicoles et a) A network of laboratories involved in the Dr A Hay Porcines (AFFSA Ploufragan) National Institute of Medical PB 53 laboratory diagnosis for swine influenza Research Les Croix viruses in different European Member States Division of Virology/WIC 22440 Ploufragan The Ridgeway France was established. Because participating Mill Hill London NW8 1AA Tel: +33 29 60 16 222 laboratories standardised the methods for UK [email protected] Tel: +44 20 89 59 36 66 ext Dr J C Manuguerra antigenic subtyping and for further genetic 2141 Institut Pasteur Dépt de characterisation, new variants can be easily [email protected] Virologie recognised. Dr Michel Bublot Unité de Génétique Discovery Research, Merial Moléculaire des Virus b) If future funding is available, the network Virology Dept Respiratoire will provide valuable information about the 254 Rue Marcel Mérieux 25 Rue du Docteur Roux 69007 Lyon 75724 Paris prevalence of swine influenza viruses. This France France information is important because swine [email protected] Tel: +33 14 06 13 354 Dr P Lenihan [email protected] influenza viruses can potentially contribute to Central Veterinary Research Dr J Heldens the evolution of new pandemic influenza strains Laboratory Intervet International BV in mammals including humans. Virology Division Virological Research Dept Abbotstown Castlenock Wim de Korverstraat 35 c) The emergence of a new H1N2 pig subtype 15 Dublin 5830 AA Boxmeer that is not covered by current vaccines was Ireland The Netherlands Tel: +35 31 60 72 694 Tel: +31 48 55 85 232 demonstrated. [email protected] [email protected] ESNIP European Surveillance Network for Influenza in Pigs 2

Acronym: ESNIP 2 EC contribution: €300 000 Starting date: 01/01/2006 Key words: Influenza, swine, surveillance, Duration: 36 months Instrument: Coordination Action evolution, diagnosis, public health risk

Summary: Problem: influenza viruses or human-avian reassortants The European Surveillance Network for Influenza During the last decade, the epidemiology of may transmit to humans via the pig. However, in Pigs (ESNIP) 2 will maintain and expand a swine influenza in Europe has become particularly the true public health risk of the pig remains surveillance network that was established during complex. At least 3 SIV subtypes – H1N1, H3N2 unknown, because there is no screening of pigs a previous EC concerted action (ESNIP, QLK2-CT- and H1N2 – are currently circulating and new for influenza virus from other hosts. 2000-01636). Three work packages (WP 1, 2, 3) reassortant viruses between these subtypes have aim at a better understanding of the epidemiology been occasionally detected. The heterogeneity Aim: and evolution of swine influenza virus (SIV) in in swine influenza has important implications The strategic objectives of the current different European countries, through an organised for the diagnosis and control. Indeed, the strains Coordination Action are: surveillance programme together with antigenic/ used in serodiagnostic tests need to be matched a) to further expand our knowledge of the genetic characterisation using standardised to the current epidemic viruses, and inactivated epidemiology and evolution of swine influenza methodology. These data will be used to improve the SIV vaccines should contain all of the prevailing viruses in Europe and to apply this knowledge diagnosis of SIV by updating the reagents used in subtypes for a broad protection. Two significant to optimise diagnostic techniques for swine classical techniques (WP 4) and by the development difficulties, however, are that there is little influenza; of a rapid, molecular test for SIV detection (WP 5). surveillance for SIVs and that our understanding b) to provide insights into the public health risk of the SI epidemiology is far from complete. of influenza in swine by monitoring swine for The virus bank and electronic database that were avian influenza viruses and by comparison established during ESNIP will also be expanded So far there have been rather limited attempts of influenza viruses in swine and in human with relevant SIV isolates and information (WP 6). at detailed characterisation of SIVs. In addition, populations. Beyond this, we will be better able to define the there has been confusion about the extent public health risks of influenza in swine. This will of antigenic evolution of these viruses, i.e. The research objectives can be grouped into six be achieved by serological monitoring of swine for considerable antigenic drift of H1N1 and H3N2 major tasks: avian influenza viruses (WP 7) and by comparison SIVs was reported in some regions of Europe a) To keep track of major changes in the of the influenza viruses that are currently (de Jong et al. 1999, 2000), but not in others epidemiology of SIV in Europe. circulating in swine, avian species and humans (Campitelli et al. 1997). One difficulty was a lack b) To further study the extent of antigenic and (WP 8). ESNIP 2 represents the only organised of standardised reagents and protocols for the genetic evolution of SIVs. surveillance network for influenza in pigs and the subtyping and antigenic/genetic characterisation c) To improve the diagnosis of SIV. first attempt to establish formal interactions with of SIV, which greatly contributed to discordant d) To expand the SIV bank and electronic human and avian surveillance networks. results between different laboratories. database which were established during ESNIP. e) To screen European swine populations for the These initiatives and interactions are entirely Virus isolation followed by haemagglutination circulation of avian influenza viruses. consistent with improved pandemic preparedness inhibition (HI) and neuraminidase inhibition (NI) f) To compare the influenza situation in swine and planning for human influenza. The with reference sera is the standard procedure for with that in humans and birds and ensure consortium consists of 11 participants who are subtyping SIVs. The HI test is also most widely dissemination of information to human and actively working with SIV. Eight participants are used for the serologic diagnosis and for serologic avian influenza researchers. from seven different EU Member States, five of studies of SIV. The reagents used in such tests which have been actively involved in the ESNIP must be updated if relatively minor antigenic network whilst one participant is from Bulgaria, changes in the circulating SIVs should occur, and Expected results: a candidate Member State (at the time of the failure to do so may result in incorrect subtyping Scientific: project start date). Participation from third of SIVs and false seronegatives in the HI test. a) Maintenance of the surveillance network countries (Hong Kong, USA) will facilitate greater There is also a strong need for alternative, more for various influenza A subtypes in swine in global interaction and worldwide understanding rapid and simple tests for detection and subtyping Europe. of the epidemiology of SIV. Comparisons of swine of SIVs. b) Data about the prevalence and circulation populations in Europe and in southern China, the patterns of different SIV subtypes in swine in classical epicentre for influenza, for the circulation Finally, pigs are known to be susceptible to Europe and comparison with the situation in of avian influenza viruses will prove invaluable infection with both human and avian influenza southern China and in the USA. 52 and permit technology and knowledge exchange. viruses and there are concerns that avian c) Antigenic and genetic characterisation of SIVs strategies on individual swine farms. In addition, addition, In farms. swine individual on strategies vaccination of design arational for is essential This SIVs. of subtyping the SIand of diagnosis the facilitate and improve will project The million. £60 of annum per UK the in loss atotal to equivalent pig, per £7 approximately at estimated been has alone SIV to due pigs fattening in gain weight reduced from resulting loss financial the UK, the In swine. fattening in outbreaks disease respiratory acute the of 50% to up in isinvolved SIV that shown have Belgium and Netherlands the in Investigations industry. swine the for disease important economically SIisan EU. the Indeed, throughout health swine of status ahigh to contribute will it because policy, health EU animal the supports Action Coordination this context, abroader In P support: policy and Technical e) Formal interaction between the SIV SIV the between interaction Formal e) aserologic of validation and Development d) SIvirus the of expansion and Maintenance c) novel, of validation and Standardisation b) reagents of type the about Recommendations a) influenza avian of prevalence the about Data d) applications otential viruses in swine in Europe. in swine in viruses countries. European different in herds pig in networks. human and avian and network surveillance swine. in viruses influenza avian for assay database. electronic and bank swine. in viruses influenza of characterisation and diagnosis the for tests rapid more SIV. for tests diagnostic classical in used be to : farmers. swine of profitability the and swine of welfare the to contribute will 2project ESNIP the way this In required. be may composition strain vaccine the in changes whether indicate will This strains. vaccine with comparison in characteristics antigenic their and Europe in circulating currently are that subtypes SIV the of picture aclear obtain will we with such potentially hazardous viruses. hazardous potentially such with contaminated be to meat pig for risk potential isa there whether reveal will which Europe, in swine in viruses influenza avian of prevalence the determine We will safety. and quality food on impact an have will research Finally, our namely: missions, OIE the with line in also are project the of objectives The c) to provide expertise and promote international international promote and expertise provide to c) veterinary disseminate and analyse collect, to b) animal of transparency the guarantee to a) solidarity for the control of animal diseases. animal of control the for solidarity diseases; eradicate and control to used methods the improve to States Member help to information scientific worldwide; status disease [email protected] Belgium Merelbeke B-9820 133 Salisburylaan Virology of Medicine Laboratory Veterinary of Faculty University Ghent R Van Kristien Coordinator: eeth Unité d’Epidémiologie et Bien- et d’Epidémiologie Porcines Unité et Avicoles de et Recherches d’Etudes Laboratoire Aliments des Sanitaire Sécurité de Française Agence Madec François [email protected] Italy Parma 43100 13/A Mercati dei Via Parma di Sezione Diagnostica Romagna dell’Emilia e Lombardia della Sperimentale Zooprofilattico Istituto E [email protected] Netherlands The Lelystad AA 8203 2004 Box O. P. CIDC-Lelystad Virology of Dept CIDC-Lelystad Loeffen Willie [email protected] UK 3NB Surrey KT15 Addlestone Haw New Lane Agency Woodham Laboratories Veterinary I Partners: [email protected] 1AA NW7 London Hill Mill Ridgeway The Medical for Research Institute National Matrosovich Mikhail [email protected] France Ploufragan 22440 53 BP Croix Les Zoopôle Porcin Etre an Brown an manuela Foni manuela [email protected] [email protected] Bulgaria Sofia 1606 Blvd “P.Slavejkov” 15 Diseases Exotic of Dept Institute Veterinary Research Diagnostic National I [email protected] Spain Gerona 17170 s/n Amer Selva La A S Avenida HIPRA Laboratorios Services DIAGNOS Diagnostic Veterinary Maldonado Jaime [email protected] France Lyon 69007 Mérieux Marcel Rue 254 Dept Merial Virology Research, Discovery Bublot Michel [email protected] USA 53706 WI Madison Drive Linden Sciences 2015 Pathobiological Medicine of Dept Veterinary of School Madison Wisconsin- of University WO Christopher [email protected] China SAR Kong Hong Pokfulam Peiris Malik vaylo Chenchev vaylo lsen

ESNIP 2 SSPE-CT-2005-022749 www.esnip.UGent.be European Influenza Surveillance Scheme

Acronym: EISS EC contribution: €1 439 790 Starting date: 1/09/2003 Key words: Surveillance, influenza, epidemiology, Duration: 36 months Instrument: Public Health Programme virology

Summary: Problem: Aim: The European Influenza Surveillance Scheme Influenza is a major public health problem in The general aim of the EISS project is to contribute (EISS) publishes a weekly surveillance report on Europe. It is associated with higher general to a reduction in the burden of disease associated influenza activity during the winter. The report practice consultation rates, increased hospital with influenza in the EU Member States. covers a total population of 484 million people admissions and excess deaths. It must also be in 30 countries and is based on data reported by considered in terms of health care planning, roughly 13 000 sentinel physicians. increased days lost due to absence from work Expected results: and influenza pandemic planning. Worldwide The collection and exchange of timely information EISS also operates the Community Network of influenza pandemics in the twentieth century on influenza activity, which will help contribute Reference Laboratories for Human Influenza in occurred in 1918-20 (more than 20 million to the annual determination of the influenza Europe (CNRL). The CNRL includes 38 laboratories deaths), 1957-58 and 1968-70. vaccine content and European influenza pandemic across Europe and its objective is to provide high preparedness activities. quality reference services for human influenza surveillance, early warning and pandemic preparedness. Potential applications: The data provided by the EISS can be used in a number of ways: a) To provide information about influenza to health professionals and the general public. b) To contribute to the annual determination of the influenza vaccine content. c) To support health care planning associated with seasonal influenza activity (e.g. hospitals and nursing homes). d) To establish surveillance structures in preparation for a pandemic.

54 www.eiss.org/cgi-files/wiw_members_display.cgi see list, adetailed For data. surveillance epidemiological and virological their for institute acontact has country Each Switzerland. and Norway plus EISS, in participate States Member EU 26 Partners: Research [email protected] Services for Health Institute Netherlands Paget John Coordinator:

EISS 2003204 www.eiss.org Biology

56 It is becoming increasingly clear that the distinction between ‘basic’ and ‘applied’ research is an outdated classification system with little relevance in today’s health sciences – and research into the biology of influenza viruses and influenza infection is a prime example of this paradigm. Research that addresses a number of fundamental questions about the molecular and structural biology of these viruses is crucial for the development of advanced solutions to a number of very practical problems in the prevention and treatment of influenza or in public health efforts to stop an epidemic.

The dysregulated innate immune response of the host is increasingly being recognised as a key factor in the severity of the clinical disease. While many of the projects assembled in this chapter include some research on this important aspect, the detection of virus and host determinants of this non-specific response is the major focus of the recently launched FLUINNATE project. EUROFLU, FLUPATH and INN- FLU all tackle – in addition to the innate immune response – several other issues related to the pathogenicity of viruses: They examine the important question of how easily viruses are transmitted from one species to another, including the crucial jump from birds to humans, and which factors are responsible for this species-specificity as well as for the clear tropism of the virus for only a very limited range of cell types within a given organism. Related to this question are studies of virus-host receptor interactions, the contribution of haemagglutinin and neuraminidase proteins to virulence in different species as well as research on cellular mechanisms that regulate the viral replication within the infected cell.

FLUPOL (targeted exclusively on influenza) and VIZIER as a broad- based effort aimed at a number of RNA-viruses (but including influenza as well as other medically relevant viruses), both aim to characterise the structure of core enzymes of the viral replication machinery with a view to finding new targets for antiviral drugs. The RespViruses consortium takes a different approach and studies the innate and acquired immune response of elderly patients to known and newly discovered respiratory viruses also with the goal of developing innovative therapeutics, such as antiviral siRNA-based approaches and strengthening the host’s immune response. Host-specific variants of the influenza virus replication machinery

Key words: Influenza virus polymerase, viral replication, X-ray crystallography, cryo-electron Acronym: FLUPOL microscopy, host-dependent mutations, EC contribution: €1 973 450 Starting date: 01/01/2007 ribonucleoprotein particle, high throughput Duration: 36 months Instrument: STREP screening

Summary: Problem: Aim: We aim to understand how the influenza virus Currently circulating H5N1 avian influenza viruses We will focus on determination of the atomic replication machinery adapts during interspecies are lethal to man and could cause a devastating structure of polymerase domains as well as transmission and to use this knowledge to pandemic if they became transmissible between the complete trimeric complex by state-of-the- provide new tools to combat potentially pandemic humans. It is therefore crucial to understand art methods such as X-ray crystallography, influenza outbreaks. the mechanisms whereby influenza virus adapts nuclear magnetic resonance and cryo-electron from avian to human hosts. The best understood microscopy. This will provide the detailed factors in inter-species transmission are certain framework required to understand polymerase characteristics of the surface glycoprotein, function and the effect of specific point mutations haemagglutinin. However, several recent in inter-species adaptation. studies have highlighted the importance for transmissibility of mutations in the proteins of We will also undertake biochemical, cellular the viral replicative machinery, in particular the and animal functional studies of the replication polymerase which transcribes and replicates machinery and identification of host cell factors the viral RNA. We propose a comprehensive interacting with the polymerase using advanced study of the molecular structure and function of functional genomics methods. In parallel, the influenza virus polymerase with the aim of candidate mutations that may be important for understanding how it adapts during inter-species inter-species transmission and virulence will be transmission. identified by bioinformatics analysis of influenza genome sequences, updated with sequences of new H5N1 isolates, as well as from studies of laboratory strains adapted from one host to another (e.g. avian to mouse).

Coordinator: Dr Stephen Cusack European Molecular Biology Laboratory Grenoble Outstation 6 Rue Jules Horowitz 38042 Grenoble France Tel: +33 47 62 07 238 [email protected]

Partners: Prof. Rob Ruigrok University Joseph Fourier Laboratoire de Virologie Moléculaire et Structurale Grenoble France [email protected] Dr Vincent Lotteau INSERM Unit 503 Prof. Hans-Dieter Klenk Lyon Institute of Virology France University of Marburg [email protected] Germany Dr Juan Ortin [email protected] National Biotechnology Centre Dr Alan Hay (CNB) National Institute for Medical CSIC Research Madrid London Spain UK 58 [email protected] [email protected] interactions. factor cell polymerase-host or polymerase targeting compounds anti-viral of development for strategies new of elaboration to lead of evolution of model acomprehensive of elaboration to contribute potentially will modelling mathematical and sequences viral of analysis bioinformatic asystematic with combined studies, these All transmission. inter-species with associated NP and polymerase in mutations of effects the assess to efficiency) replication activity, based use We will genome. influenza the of replication and/or transcription for required factors host identify will we complexes of tagging and method two-hybrid yeast Using mutations. specific species of interpretation astructural provide We will microscopy. cryo-electron by complex nucleoprotein RNA- polymerase-viral the of structure the elucidate We will structures. atomic their determine and (NP) nucleoprotein and subunits polymerase of domains soluble and folded independently identify systematically We will Ex results pected in vivo in characterisation (e.g. polymerase polymerase (e.g. characterisation in vitro in : in vivo vivo in and cell- and particular the structure of the target active sites, sites, active target the of structure the particular in function, polymerase of understanding based structure adetailed of lack However, the proteins. cell host in found those from different significantly be to likely sites active functional several contains and replication viral for essential is it since drugs, anti-viral new for target excellent isan complex polymerase virus influenza The P diagnostic purposes. diagnostic for used be could which antibodies monoclonal specific as such tools biology molecular new include applications Other compounds. anti-viral for screen to assays high-throughput new develop to aim we results structural and biochemical our on Based proteins. partner cell host with interactions including machine, complex this of action of mechanism the of detail resolution atomic providing by situation, this rectify to aim polymerase on studies functional and structural Our direction. this in progress hindered has applications otential :

FLUPOL SP5B-CT-2007-044263 Innate immunity in influenza virus infection of mammalian airways

Key words: Emergent diseases, emerging influenza Acronym: FLUINNATE A virus, viral pathogenicity factors, interferon, EC contribution: €1 436 130 Starting date: 01/01/2007 innate immunity, human airway epithelium, host Duration: 36 months Instrument:STREP range, antiviral response

Summary: regularly, often leading to excess mortality in segment reassortants will be produced and fully Emerging influenza A virus (FLUAV) infections susceptible populations. In addition, influenza characterised together with the parental strains pose a considerable health threat to mankind. A viruses can cause devastating pandemics in with respect to growth kinetics in tissue culture The molecular determinants governing increased humans. An avian influenza A virus originating and in vivo, the capacity to induce or respond to virulence of emerging virus strains in humans are from Asia and currently circulating among interferon, and the capacity to induce disease or presently not well understood. FLUINNATE proposes domestic birds in Europe and neighbouring death in experimental animals. The technology to identify and study the essential viral and host countries makes headlines because of its potential for expressing, purifying and analysing the viral factors that determine the outcome of infection. to infect and kill people. If further adaptation to RNA polymerase complex will be established FLUAV enters the human respiratory tract and must humans occurs, this virus strain might become the and further refined, as well as biochemical and replicate in the face of multiple innate immune origin of a future pandemic. biophysical approaches to identify co-purifying defence mechanisms to establish infection in vivo. host cellular factors. Advanced tests for protein- Although influenza viruses belong to the best protein interactions such as the yeast three- Successful viruses must adapt to intrinsic cellular studied viruses, the host adaptation processes hybrid system will be set up and candidate restriction factors and evolve the capacity of which enable influenza viruses to jump from interactors evaluated in functional tests, based on circumventing the antiviral interferon (IFN) response, one species to another are largely unknown. transfection experiments. The real-time RT-PCR either by limiting IFN production or by blocking IFN Likewise, the properties required for host-to- methodology will be optimised for the analysis of actions. We will test the hypothesis that the speed and host transmission are presently not understood. interferon and cytokine responses and gene array efficiency by which a given virus circumvents these early Moreover, efficient control of influenza virus data will be generated. host responses are critical determinants in its host range infections is still not possible. Immunisation and pathogenicity. In this respect, the crucial role of regimes are continually being confronted with the virus polymerase and its cellular interactors will be the extreme antigenic variability of influenza A Expected results: analysed. Likewise, the importance of the IFN-inducible viruses brought about by antigenic drift and shift. • Generation of reassortant influenza viruses Mx GTPase as a major anti-FLUAV effector molecule It is evident that new approaches and reagents to containing genes from high and low virulent will be evaluated. control influenza are urgently needed. PR8 strains by reverse genetics. • Generation of recombinant influenza viruses Virus-induced inflammatory cytokines and expressing various tagged polymerase chemokines exert powerful effects against FLUAV Aim: complexes. in lung epithelial cells. However, they may be The FLUINNATE objectives focus on the • Generation of recombinant viruses with distinct detrimental to the host, causing accelerated identification of influenza A virus genes and receptor specificity. influenza pathogenesis in the human respiratory gene products which contribute to virulence/ • Identification of viral gene constellations tract. We will analyse viral factors governing the pathogenicity in experimental animal and tissue responsible for high pathogenicity in Mx1+/+ innate antiviral cytokine response and determine the culture models. The required animals are available mice. impact of these factors on virus growth, cell survival and will consist of mice with the wild-type Mx1 • Sequence comparisons of newly isolated and pathogenicity. Human, avian and porcine gene as part of the full innate immune response, H5N1 viruses with emphasis to the polymerase FLUAV will be used in animal models and in cell various strains with targeted mutations in specific subunit PB2, the nucleoprotein NP, and the culture systems, such as human airway epithelium. genes and pigs as natural hosts and ‘mixing interferon-antagonistic protein NS1. The present studies should generate important vessels’ for influenza A viruses. In addition, • Optimisation of the co-immunoprecipitation information that will help to better understand human airway and porcine epithelial cell cultures assay using recombinant influenza viruses the processes involved in the emergence of lethal will be established and characterised. Human expressing a tagged polymerase complex. influenza viruses and to develop efficient control airway epithelial cell cultures are a rare but • Construction and expression of polymerase measures against these devastating pathogens. most precious substrate to study the biology of clones from human and avian influenza viruses. influenza virus infection. • Testing the functionality of these polymerase clones in human and avian cells. Problem: The influenza viruses used will be human, • Reconstitution of active viral polymerase Influenza is a highly contagious, acute respiratory avian and swine strains, some of which will complexes from subunits. illness that is still a major health problem. be generated by reverse genetics entirely from • Set-up of a yeast two-hybrid systems with viral 60 Epidemics caused by influenza A viruses occur plasmids. Stock viruses and single and multi- polymerase subunits as a bait. • Detection of PTX3 levels in infected mice. mice. infected in levels PTX3 of • Detection virus influenza of spectrum the of • Assessment viruses influenza avian different of • Comparison from • Results and induction IFN of data • Comparative • receptor viral of consequences the on • Data epithelial airway human of data array • Gene host polymerase-interacting of • Identification aco- using by factors host for • Screening yeast in system athree-hybrid of • Establishment In vivo In purification strategy followed by identification identification by followed strategy purification PB1using +PB2PB1 or +PA abait. as recognition by PTX3 and other pentraxins. pentraxins. other and PTX3 by recognition cells. porcine in IFN to susceptibility and induction IFN for viruses influenza swine with cells. porcine in virus influenza swine by induction IFN of basis viruses influenza swine different for IFN to susceptibility virus. influenza swine with infection an during IFN of role exact the NK-cells). neutrophils, T- cells, B-lymphocytes, and dendritic (macrophages, cells immunocompetent human of function and expression gene for specificity infection. virus influenza upon cells virulence. enhanced and activity polymerase high in involved factors spectrometry. mass by factors host co-purifying of pathogenesis studies in pigs to examine examine to pigs in studies pathogenesis in vitro in in vivo in studies on the molecular molecular the on studies in a porcine system. system. aporcine in may have an impact on future pandemic planning. pandemic future on impact an have may and authorities healthcare and epidemiologists to interest great of be will determinants virulence viral and factors restriction host antiviral on Results purposes. research for community scientific the to provided and produced be will viruses recombinant useful systems) genetics reverse as (such technology cutting-edge Using agents. therapeutic possibly and markers new candidate as well as infection influenza of immunopathology the on vistas innovative provide will FLUINNATE that isexpected It P • Susceptibility of TLR-deficient mice to avian avian to mice TLR-deficient of • Susceptibility influenza to mice D6-deficient of • Susceptibility virus influenza critical the of • Identification to mice targeted gene PTX3 of • Susceptibility • Formation of a network for collaboration on on collaboration for anetwork of • Formation applications otential virus infection. infection. virus infection. virus influenza to mice TIR8-deficient of Susceptibility PTX3. by recognised components infection. virus influenza institutions, agencies and hospitals. hospitals. and agencies institutions, research Chinese with viruses influenza avian viruses. influenza : Scholars of the University of of Oxford University the of and Scholars Masters Chancellor, The D uni-marburg.de 166 65 Mikhail.Matrosovich@staff. 28 21 64 +49 Tel: Germany Marburg 35032 10 Marburg Biegenstrasse Philipps-Universität D 69 [email protected] 73 64 92 +32 Tel: Belgium 25 Gent 9000 Sint-Pietersnieuwstraat Gent Universiteit R Van Kristien Prof. 36 34 [email protected] 24 77 05 +39 Tel: Italy Siena 53100 1 Fiorentina Via Srl and Diagnostics Vaccines Novartis D Partners: 34 65 [email protected] 12 76 +49 Tel: Germany Freiburg 79098 Fahnenbergplatz Freiburg Albert-Ludwigs-Universität O Prof. Coordinator: Tel: +44 18 65 27 55 80 55 [email protected] 27 65 18 +44 Tel: UK 2JD OX1 Oxford Square Wellington Offices University r E Matrosovich r Mikhail r S tefania Crotta tefania rvin Fodor rvin tto Haller tto eeth D 27 19 [email protected] 85 63 21 +86 Tel: China Shanghai 20031 Road Sciences Yueyang of 320 Academy Chinese Biological of Sciences Institute Shanghai S Bing Prof. humanitas.it 48 24 fondazione.humanitasricerca@ 24 82 02 +39 Tel: (Milan) Italy Rozzano 20089 56 Manzoni Via la per Ricerca Humanitas Fondazione A Prof. 811 88 56 14 +33 Tel: France Paris 75724 Roux Dr du Rue 25 Pasteur Institut D Tel: +39 05 77 24 35 08 35 [email protected] 24 77 05 +39 Tel: Italy Siena 53100 1 Fiorentina Via Srl Alta [email protected] r Maria Paola Cesaroni Cesaroni Paola r Maria r N adia N adia lberto Mantovani lberto affakh un

FLUINNATE SP5B-CT-2006-044161 www.fluinnate.org Molecular factors and mechanisms of transmission and pathogenicity of highly pathogenic avian influenza virus Acronym: EUROFLU Key words: Avian influenza viruses, pathogenicity, EC contribution: €1 355 811 Starting date: 01/01/2007 transmission, replication, virulence, vaccines, Duration: 36 months Instrument: STREP drugs, diagnosis

Summary: Aim: Epidemics caused by highly pathogenic avian pathogenicity within organisms. The in vivo The overall objective of the EUROFLU consortium influenza viruses (HPAIV) are a continuing threat characterisation of immune responses and is to understand the molecular determinants to human health and to the world’s economy. This investigation of mechanisms of neurotropism and the mechanisms responsible for virulence, now becomes clearly evident after the emergence in chicken and in mice upon HPAIV infection host specificity, inter-species transmission and of the HPAIV H5N1 subtypes that have infected will allow the correlation and verification of pathogenicity of HPAIV, which will eventually and killed humans in Asia and Europe. The global EUROFLU’s experimental and computational accelerate the development of effective diagnostic dimension of current HPAIV infections of birds results in natural and in experimental host tools, anti-HPAIV vaccines and drugs. and humans highlights an urgent need to increase species, respectively. To achieve this goal an interdisciplinary international and multidisciplinary research consortium of experts in virology, immunology effort collaborations to develop new diagnostics, and bio-computing has been established. vaccines and drugs. Problem: EUROFLU will provide European authorities, Influenza is currently considered one of the policy makers and the community with advanced The EUROFLU consortium integrates most severe threats to human health and animal scientific knowledge supporting a successful interdisciplinary experimental and computational welfare. During the past 10 years outbreaks of control of avian influenza. The specific research research approaches carried out by 11 partners HPAIV have occurred mostly in Central and East aims are: from four EU Member States and from one Asia, and since 1999 these viruses have also been Associated Member State. The overall objective found in the Middle East as well as in Eastern a) Defining virus/cell surface-interactions: of EUROFLU is to study the molecular factors and Southern Europe. The unprecedented spread This section of the scientific work aims to and mechanisms of HPAIV transmission and of the H5N1-type HPAIV in poultry and wild characterise the specific interaction between pathogenesis. In order to build a profound waterfowl throughout Asia, Europe and more domains of the haemagglutinin subunit 1 scientific platform that will help to support the recently in West Africa, has been associated with (HA1) and the host-cell receptor (termed as European policy makers in the fight against at least 241 transmissions to humans with a case recognition and targeting markers (RTMs)) as HPAIV, EUROFLU will focus on three major fatality rate (CFR) of more than 50%. The virus one of the main factors determining host range. research tasks: also spread to several other mammalian species This task will be approached by advanced including cats. Should these viruses acquire the computational and biochemical analysis. Focus a) Identifying, characterising and validating capacity for easy human-to-human transmission, will be on human and avian isolates of the HPAIV factors that are involved in the the outbreak of a new influenza pandemic with a H5 and H7 types of HPAIV and results will recognition and targeting of the virus to the considerably high CFR could be expected. be compared with data gained from human cellular host receptor, thereby defining host H1 and H3 virus strains including the 1918 range through advanced computational and Several infections of humans with the recent ‘Spanish Flu’. biochemical analyses. Special focus will be HPAIV H5N1 viruses have been characterised by put on the viral haemagglutinin protein, which an unusually strong cytokine response in the host b) Analysing intra-cellular virus/host-interactions: binds to sialic-acid coupled cell receptors of (‘cytokine storm’) that appears to contribute to viral This part of the projects aims to understand the human and avian hosts. pathogenicity. There are also first indications of a molecular mechanisms of interaction b) Revealing viral and cellular factors and neuro- and an endothelial-tropism of recent HPAIV in between HPAIV and host-cell factors and the mechanisms that regulate virus replication mammals. The molecular basis of the latter findings role of these interactions for viral within the infected cell (and which can that may determine virulence and host cell tropism is pathogenesis. The analyses will involve reverse therefore determine cell tropism and host not known and needs to be urgently defined. genetics, genomic and proteomic specificity) employing diverse virology, techniques and will elucidate the similarities molecular- and cell-biology methods. Focus To date there is no vaccine against H5N1 and and differences of how HPAIVs replicate in will be on virus/host interactions between viral variants resistant to current antiviral drugs are avian and mammalian host cells and how they factors of HPAIV and their crosstalk to cellular emerging. This highlights the urgent need for fast evade cellular defences. Furthermore, the factors. and specific diagnosis, an effective vaccine and current knowledge on HPAIV’s misuse of c) Using chicken and mouse models for in vivo new efficient antiviral drugs. cellular activities to support their propagation analysis of HPAIV infections of birds and will be extended. 62 mammals to monitor HPAIV transmission and Coordinator: Paul-Ehrlich-Strasse 28 84505 Bratislava 45 c) Determining virus/host-interactions in animal Dr Stephan Pleschka 72076 Tübingen Slovakia hosts. The studies in this section of the project Justus-Liebig-Universität Germany Tel: +42 12 59 30 24 18 Institute for Medical Virology Tel: +49 70 71 96 72 54 [email protected]

will use mouse and chicken model systems for: euroflu Frankfurter Strasse 107 [email protected] Dr Joachim Klein 69978 Tel Aviv / (i) the characterisation of immune responses 35392 Giessen Prof. Ursula Dietrich RiNA Network Israel Germany Georg-Speyer-Haus Taku Strasse 3 Tel: +97 23 64 06 709

after HPAIV infection (ii) the investigation de

Tel: +49 64 19 94 77 50 Paul-Ehrlich-Strasse 42-44 14195 Berlin [email protected] . of mechanisms of viral neurotropism and [email protected]. 60596 Frankfurt Germany Prof. Maria Sakarellos-Daitsiotis neuropathogenicity in mammals and in birds. uni-giessen.de Germany Tel: +49 30 84 41 66 33 University of Ioannina Tel: +49 69 63 39 52 16 [email protected] Dept of Chemistry Partners: [email protected] Dr Thorsten Wolff 45110 Ioannina Dr Ayub Darji frankfurt.de Robert Koch Institute Greece Centre de Recerca en Sanitat Dr Rachel Kreisberg-Zakarin Nordufer 20 Tel: +30 26 51 09 83 86 Expected results: Animal IBEXPERTS Ltd giessen

D-13353 Berlin [email protected] - a) VIRUS/CELL SURFACE INTERACTIONS: Campus de la UAB 74 Rambam Street Germany Prof. Stephan Ludwig The expected outcome is enhanced knowledge Bellaterra 43602 Ra’anana Tel: +49 30 45 47 22 78 Zentrum für Molekularbiologie 08193 Barcelona Israel [email protected] der Entzündung uni of HPAIV cell specificity and tropism by the Spain Tel: +97 29 74 11 769 Prof. Nir Ben-Tal Von-Esmarch-Strasse 56 . identification and characterisation of RTMs and Tel: +34 93 58 14 558 [email protected] Tel Aviv University 48149 Münster [email protected] Prof. Rudolf Toman The George S Wise Faculty of Germany through the investigation of HPAIV-RTM/host Prof. Oliver Planz Institute of Virology Life Sciences Tel: +49 25 18 35 77 91 cell receptor interaction. This will lead to the Friedrich-Loeffler Institute Dubravska Cesta 9 Dept of Biochemistry [email protected] www identification of viral and cellular structures that can be used for diagnosis and for the generation of new vaccines and might be interesting targets for drugs designed to block virus/cell interaction. b) INTRA-CELLULAR VIRUS/HOST INTERACTIONS: This analysis is expected to increase the understanding of: (i) HPAIV replication in avian and in mammalian host cells; (ii) the viral mechanisms aimed at evading cellular defences; (iii) the HPAIV usage of cellular mechanisms for their propagation.

The knowledge gained will allow the characterisation of viral conditions that define cell tropism (avian or human) of HPAIV and will also pave the way for new anti-HPAIV drugs interfering with virus replication. c) VIRUS/HOST INTERACTIONS IN ANIMAL HOSTS: The results of this analysis will elucidate basic virulence mechanism(s) in the studied mouse and SP5B-CT-2007-044098 chicken models and they will also give important The EUROFLU STREP is composed of four Work Packages (WP1-4) and four Scientific Teams insights into the course of H5N1 infections in (Computational, Virology, Biochemical and Model Organism). The management in WP4 is headed by the Coordinator who will be assisted by the Administrative Manager and a Management Board, which humans, which will help to adjust the medical includes members of the consortium. Each WP is headed by a Work Package Leader (WPL), who will treatment of patients. implement the scientific and organisational decisions of the management. The scientific teams will work on the indicated topics and will strongly interact within the consortium.

WPA Potential applications: Management Results gained in the EUROFLU project will provide the knowledge base for new therapeutic approaches in the development of improved WP1 leader WP2 leader WP3 leader diagnostic tools and of vaccines to block HPAIV infections. It will set the molecular basis for WP1 WP2 WP3 Identification of viral and Study of virus/host- HPAIV pathogenesis and new anti-viral drugs against influenza viruses cellular RTMs used for interaction in neurotropism study in binding of avian/human pathogenesis and independent of the specific virus strain and chickens and mice circumvent the constant risk of selecting resistant IV to the cell transmission Model Organism Team viral variants that make current drugs ineffective. Computational Team Virology Team The new understanding of virulence mechanisms will help to develop new strategies for the treatment of patients. WP2 Analysis of viral NS1, HA and NP proteins. WP1 Analysis of cellular WP3 Validation of viral and proteins Validation of viral and cellular RTMs through cellular RTMs through in vitro RTM/host cell Computational Team in vitro RTM/host cell receptor binding assay receptor binding assay

Biochemical Team Biochemical Team EUROFLU Avian influenza: impact of virus-host interactions on pathogenesis and ecology

Key words: Avian influenza virus, virus-host Acronym: FLUPATH interactions, virus-receptor interactions, host- EC contribution: €1 915 800 Starting date: 01/01/2007 specificity, expression profiling, virulence, Duration: 36 months Instrument: STREP pathology, ecology, innate immunity

Summary: Problem: Aim: Highly pathogenic avian influenza viruses (HPAIV) Lack of sufficient fundamental knowledge: avian This proposal aims to generate data on significant have acquired the unprecedented and alarming influenza represents one of the major concerns issues linked to AI outbreak management on capacity to infect humans. By establishing a for public health that has recently emerged from which scientific knowledge is currently lacking. permanent ecological niche in wild birds, HPAIV the animal reservoir. The increased relevance These issues are all related to virus-host will pose a continuous risk for poultry and fatal of avian influenza in the fields of animal and interactions. Four major tasks (work packages) human infections, especially if these birds excrete human health has highlighted the lack of scientific have been identified which address the objectives HPAIV without showing any clinical signs of information on the disease. This has hampered of task 3 of the EU call SSP5-B INFLUENZA: disease. These changes in the ecology of the the management of some of the recent crises, ecology and pathogenesis of avian influenza disease and behaviour of the virus may create resulting in millions of dead animals, concern infections: opportunities for a pandemic virus to emerge. over loss of human lives and management of the virus’s pandemic potential. For this reason, and Work Package 1 (WP1) addresses the issue Attempts to avoid or contain HPAIV outbreaks have for the devastating effects on the poultry industry, of pathogen-host interactions and virulence been largely unsuccessful. This can be directly linked international organisations such as WHO, OIE determinants at the molecular level. Little is to our lack of fundamental knowledge. Therefore, it and FAO have worked together and established a know about the host-response following infection is essential to increase our knowledge of the ecology coordinated set of guidelines and action plans to with viruses that differ in virulence and gene and pathology of avian influenza virus infections in combat the ongoing Asian epidemic. constellation. The contribution of specific viral poultry and other species. genes — or gene sequences — to pathogenesis, Due to the low profile of avian influenza until host- and tissue-tropism as well as their role in Full understanding of the ecology and pathogenesis 1997, a significant amount of information and interference with host defence mechanisms will of HPAIV requires a multi-disciplinary approach the specific tools necessary to manage avian be examined in vivo in different species in animal determining host-pathogen interactions and the role influenza epidemics adequately are lacking. This experiments as well as in vitro using professional played by the host immune response. To this end, the includes both the EU situation and the ongoing antigen presenting cells such as dendritic cells and FLUPATH consortium was established. H5N1 crisis. Recent outbreaks of HPAI have macrophages (see also WP4 below). State-of-the- affected avian species that are showing a reduced art technology such as gene-expression profiling FLUPATH is composed of 13 partners, six of susceptibility to this virus. If HPAI infection of the using DNA micro-arrays and identification of which are National Reference Laboratories for wild bird host becomes compatible with normal differentially expressed genes using gene libraries avian influenza. The consortium further includes behavioural patterns and migration, the result will obtained by suppression subtractive hybridisation five academic institutions and two institutions be the development of an endemic cycle in wild (SSH), will be used to identify specific host genes that specialise in animal science and health. The birds. The consequences of such a situation are which are involved in the reaction to infection participants, with expertise in chicken genomics, unpredictable and potentially very dangerous. by specific avian influenza virus genotypes. The micro array technology, pathology, receptors, involvement and importance of viral genes as well innate immunity and chicken immunology will use Retrospective analysis of recent outbreaks has as host genes will be evaluated by cross-validation multidisciplinary and complementary approaches permitted the identification of weak points in and in vitro experiments. to address key problems and unanswered the management system that represent areas of questions with respect to the ecology and uncertainty for which improvement is required. Work Package 2 (WP2) is mainly concerned pathogenesis of avian influenza. Several of these weak points can be directly linked with studying the ecology and pathology of to our lack of fundamental knowledge about the different avian influenza strains in different host FLUPATH will provide knowledge and tools for new importance of both viral as well as host factors in species. These studies will evaluate and compare strategies which will be tailored for the control and determining the outcome of infection. Therefore, host-pathogen interactions of different HPAI management of avian influenza at the European and it is essential to increase our effort to enhance and LPAI viruses. These evaluations include international level. This will limit the impact of the our knowledge about the ecology and pathology histopathological descriptions of virus infection, disease both in terms of human health and losses to of avian influenza virus infections in poultry and quantification of viral loads in different organs the poultry industry. The accompanying reduction in other species. and tissues by real-time RT-PCR and confocal animal slaughter and financial and economic losses immunohistochemistry. Apart from chickens, will place a significantly lower demand on EU and specific attention will be given to water birds such 64 Member States’ budgets. as ducks (mallard) which seem to be clinically less and natural killer (NK) cells. The role of the HA HA the of role The cells. (NK) killer natural and (M macrophages (DC), cells dendritic particular in system, defence innate the of compartments cellular different the with interact to and host its to adapt to virus the of requirements the analyse will studies These system. immune innate the with viruses LPAI HPAI and of interaction the on focusing transmissibility, and pathogenesis virus influenza avian determine that factors virulence of identification the with isconcerned 4(WP4) Package Work epithelium. airway human of cultures and ducks, and turkeys chickens, from explants tracheal including tissues and cells different using by studied be will transmission avian-to-human and avian-to-pig avian-to-avian, for mechanisms Molecular humans. and pigs to transmission interspecies for potential enhanced an with virus the provide might phenotypes receptor Different strains. pandemic new potentially of emergence and pathogenicity transmission, interspecies in viruses avian of activity neuraminidase and specificity receptor of role the determining of goal the with interactions virus-receptor of issue the address will 3(WP3) Package Work viruses. mammalian-adapted using by examined be will barrier species the cross to influenza avian of capacity Finally, the genetics. reverse of means by generated mutants specific or isolates natural either using system model mouse and a chicken in examined be will 627 lys PB2 mutations and NS1 the as such truncations mutations adaptive several of significance The viruses. human and avian for vessel’ a‘mixing as serve may pigs since influenza avian of transmission and maintenance in role their assess to order in conducted be will pigs on experiments Furthermore, virus. the of transmission in implicated are therefore and influenza avian pathogenic highly to susceptible Φ ) viruses will allow us to track changes in the present mammalian species. Work on currently circulating of its evolution, and its behaviour in avian and understanding of the origins of HPAIV, the patterns The FLUPATH proposal will improve our Ex investigated. be also will responses eliciting in protein this disease has had in the past. past. the in had has disease this that impact the in reduction anoticeable in result should This level. international and a European both at AI of management and control the for tailored be will which strategies new for tools and knowledge provide FLUPATH ultimately will P strategies to combat avian influenza. clearly be of high value when implemented in novel mammals, will be determined. This knowledge will which determine the transmissibility of the virus to information on critical viral and host factors, posed by HPAIV such as the current H5N1. Essential cell biological and molecular basis of the threat more complete understanding of the immunological, It is anticipated that this approach will permit a threat of a pandemic increase. situation and thus issue precise warnings, should the applications otential results pected in vivo in and and [email protected] 693 [email protected] 38 02 32 +31 Tel. Netherlands The Lelystad AA 8203 39 Houtribweg (CIDC- Lelystad) Lelystad Dierziekte Controle Instituut Centraal D Coordinator: r Ben Peeters r Ben in vitro in : cytokine cytokine : Tel: +32 92 64 73 69 73 64 92 +32 Tel: Belgium Merelbeke 9820 133 Medicine Salisburylaan Veterinary of Faculty Ghent of University D 69 43 [email protected] 08 98 04 +39 Tel: Italy (PD) Legnaro 10 35020 dell’Università Viale Virologia di Venezie Laboratorio delle Sperimentale Zooprofilattico Istituto D [email protected] 759 [email protected] 75 22 35 +61 Tel: Australia Geelong Rd 3220 Portarlington 5 24 Bag Post Laboratory Health Animal Australian Industries, Livestock CSIRO D 222 [email protected] 16 60 29 +33 Tel: France Ploufragan 22440 53 P B Ploufragan de site AFSSA Aliments des Sécurité de Sanitaire Française Agence D Partners: Tel: +44 20 88 16 25 92 25 [email protected] 16 88 20 +44 Tel: UK 1AA NW7 London Hill Mill Ridgeway The Medical for Research Institute National D 39 73 [email protected] 32 19 +44 Tel: UK 3NB KT15 Surrey Addlestone Lane Agency Woodham Laboratories Veterinary D [email protected] r r Kristien van R van r Kristien r I Jestin r Veronique r Mikhail Matrosovich r Mikhail r I T on laria Capua laria an Brown an S chat, D r John Lowenthal eeth Tel: +41 31 84 89 377 [email protected] 89 84 31 +41 Tel: Switzerland Mittelhäusern 293 3147 Sensemattstrasse and Immunoprophylaxis Virology of Institute D 57 88 [email protected] 53 19 51 +49 Tel: Germany Hannover 30559 17 Bünteweg Virology of Institute Hannover Hochschule Tierärztliche D D 18 42 [email protected] 27 15 13 +44 Tel: UK 9PS Midlothian EH25 Edinburgh and Genetics Genomics of Department Institute Roslin D 358 [email protected] 34 25 30 +31 Tel: Netherlands The Utrecht 3508 1 Yalelaan Immunology of Medicine Division Veterinary of Faculty University Utrecht D Tel. +31 32 02 38 108 [email protected] 38 02 32 +31 Tel. Netherlands The Lelystad 8200 15 Edelhertweg 65 Box PO BV ID-Lelystad D 303 [email protected] 89 94 28 +97 Tel: Israel Rehovot 76100 12 Box O. P. Sciences Animal of Sciences Dept Food Environmental and Agricultural of Faculty of Jerusalem University Hebrew The r A Herrler r Georg r D r D E van r Willem r A rtur S rtur nnemarie R nnemarie an Heller an Burt avid ummerfield ebel den

FLUPATH SSPE-CT-2007-044220 www.flupath.eu Influence of viral proteins of avian influenza virus on the innate immune response of birds

Acronym: INN-FLU Key words: Avian influenza virus, innate immunity, EC contribution: €1 800 000 Starting date: 01/01/2007 pathogenesis, susceptibility, transmission, wild Duration: 36 months Instrument: STREP bird species, monitoring

Summary: Problem: Aim: We are currently facing a continuing infection The continuing outbreaks of disease caused The principal objectives of the INN-FLU cycle caused by a highly pathogenic avian H5N1 by hpH5N1 in poultry and other birds and of consortium are to use synergies between the influenza virus (hpH5N1) in several countries in human cases in Asia, Europe and Africa are partners in order to improve knowledge about Asia. This virus may have the potential to become cause for serious concern. Unlike in the past for the susceptibility of different bird species for a pandemic virus. There have been outbreaks hpAIV, there are increased concerns about the AIV, the innate immune response in AIV infected of the H5N1 virus in poultry in several countries potential of this hpH5N1 virus to initiate a human animals and the ecology of AIV, important for in Asia since 1997. In 2005 hpH5N1 was first pandemic. It is, however, not possible to predict the transmission of the infection between wild observed outside of Asia in European poultry, with certainty if and when this might occur. and domestic birds. The work plan includes which was followed by a fast spread in Europe exclusively bird species due to the importance during February 2006. The spread of the hpH5N1 virus during the last of the disease in these species. To achieve this year was not to be predicted in the observed goal an interdisciplinary consortium of experts The most dangerous property of this hpH5N1 dimensions, but also emphasised the need for in molecular virology, genetics, immunology virus is its ability to infect and cause disease additional knowledge of the disease in poultry, and ornithology has been established. INN-FLU in other species than birds including cats, specifically water fowl. In February 2006 the will provide European authorities, policy makers tigers, dogs and stone martens. The INN-FLU virus entered the European Union with the first and the community with scientific knowledge consortium integrates nine partners from six description of hpH5N1 in Greece. Whether this enabling a better prediction of potential risks and EU Member States (Germany, Belgium, United occurred via migratory birds is not clear. a successful control of avian influenza. Kingdom, Poland, France and Greece). The project will contribute to an understanding of It is becoming increasingly evident that hpH5N1 The scientific aims of the consortium are: virus pathogenesis and virulence determinants in can spread via infected ducks and probably also chicken, determine host-pathogen interactions via other migratory species. The hpH5N1 virus a) comparing different hosts for the susceptibility and define the molecular basis for host specificity. was subsequently isolated from more stationary to HPAI H5N1 infection. Investigation on the It is based on four areas of research: living birds (e.g. swans) in many EU Member susceptibility of different bird species including a) Characterisation of the virulence of hpH5N1 in States. This raised great concern with regard to the pathogenesis, immune response and clinical several species. the potential of migratory birds for propagating picture will yield important data about the time b) Determination of host factors influenced by and spreading these viruses and it underlined of infection and disease symptoms which can be AIV infection during the very early phase of the important role of wild birds in viral spread. observed and looked at in the field. Observed infection. In addition, these H5N1 viruses become less differences in virulence between bird species can c) Research on whether different lines of chicken pathogenic to domestic ducks but remain possibly be correlated to differences in tissue breeds show differences in resistance to pathogenic to other domestic poultry. distribution of the virus (tissue tropism) and the hpH5N1 infection. local immune response; d) Investigation of contact points between wild Host factors which influence the outcome birds and domestic free range poultry and of the hpH5N1 infection and the function of b) defining the role of genetically-determined migratory behaviour of birds based on satellite viral elements which influence the virulence innate immunity in susceptibility and resistance tracking. of the virus are poorly understood. Research to HPAI H5N1 infection. The results will and technological developments will further inform us whether there are differences in Experiments will address the question of whether increase the knowledge of AIV and influenza susceptibility to hpAIV in certain chicken lines HA molecules from non-H5 and non-H7 strains virus in general. Basic research on virus structure and if this difference can be related to specific can give low pathogenic viruses the potential and function, viral pathogenesis, and the host components of the innate immune response. A to become highly pathogenic. Studies on the response due to infection will lead to more special emphasis will be laid on the interferon host-immune response will focus on the antiviral effective approaches to controlling influenza virus response during the very early phase of innate immune response in birds, the role of the infections. infection; influenza virus protein NS1 in contributing to virulence in chickens and expression of genes c) defining the influence of NS1 protein of AIV related to the interferon pathway including Mx. on the innate immune response of chickens. 66 Available data indicates that NS1 influences / eu . mechanisms in the cell which directly or Potential applications: Coordinator: Dr Thomas W Vahlenkamp, indirectly reduce the cytokine responses of Outbreaks of hpH5N1 in domestic poultry, wild

Dr Jürgen Stech flu infected hosts. As AIV field isolates with birds and humans have caused serious concerns. Friedrich-Loeffler-Institut - Federal Research Institute for truncations in the NS1 genes have been The unpredictable speed of the spread of the Animal Health

Boddenblick 5a inn

described it will be important to analyse their virus during the last years has left questions . significance. Tools to study the avian cytokine about how the virus became present in a number 17493 Greifswald - Insel Riems system have only recently become available of European countries in a very short time. The Germany and a significant number of these tools have INN-FLU project will add useful information for Tel: +49 38 35 17 172 [email protected] been established by partners of the INN-FLU surveillance studies on which birds should be www consortium; predominantly monitored and which clinical signs Partners: Prof. Dr Peter Staeheli Dr Zenon Minta can be expected. This will have great impact on University of Freiburg National Veterinary Research d) analysing the function of several AIV genes monitoring programmes and their design. The Dept of Virology Institute for their contribution to virulence in chicken. knowledge that will be gained in the INN-FLU Hermann-Herder-Strasse 11 Al Partyzantow 57 79104 Freiburg 24100 Pulawy The work is focused on the viral proteins project with regard to the molecular determinants Germany Poland haemagglutinin (HA) and neuraminidase (NA). important for innate immune response and Tel: +49 76 12 03 65 79 Tel: +48 81 88 63 051 [email protected] [email protected] Several mechanisms have been hypothesised that disease pathogenesis will improve and support Prof. Dr Bernd Kaspers Dr Didier Vangeluwe only the HA gene of the H5 and H7 can acquire vaccine developments to control AIV. Institut für Tierphysiologie Institut Royal des Sciences Veterinärstr 13 Naturelles de Belgique the property to become the highly pathogenic 80539 München 29 Rue Vautier pathotype of AIV. The aim of this task is to Germany 1000 Brussels determine whether only the introduction of a Tel: +49 89 21 80 37 58 Belgium [email protected] Tel: +32 26 27 43 55 polybasic cleavage site would be sufficient to lead muenchen.de Didier.Vangeluwe@ to the hpAIV phenotype and whether something Dr Thierry van den Berg naturalsciences.be Veterinary and Agrochemical Dr Rima Zoorob other than H5 and H7 genes can also support the Research Centre Centre National De La generation of hp AIV; Groeselenberg 99 Recherche Scientifique 1180 Brussels FRE 2937 Belgium 7 Rue Guy Moquet e) determining the ecology and transmission of Tel: +32 23 79 06 30 BP 8-94801 Villejuif avian influenza. This will involve the investigation [email protected] France Dr Colin Butter Tel: +33 14 95 83 500 of contact frequencies between wild birds and Institute for Animal Health [email protected] domestic fowl in open pens. Monitoring of wild High Street Dr Marios Georgiadis Compton Aristotle University of birds and recording of contact points between Newbury RG20 7NN Thessaloniki wild birds and domestic poultry in free-range Berks 54124 Thessaloniki UK Greece pens will yield important epidemiological data for Tel: +44 16 35 57 72 58 Tel: +30 23 10 99 99 30 future models applicable for wild life monitoring [email protected] [email protected] programmes.

Expected results: It is expected that the results of the INN-FLU project will contribute significantly to a better SSPE-CT-2006-044372 understanding of why the new hpH5N1 isolates are able to kill different wild bird species (including water fowl). Infection studies with the hpH5N1 AIV of different species of birds will answer the questions about the induction of disease symptoms as well as the virus excretion in these animals. Also the age dependency for the occurrence of disease symptoms will be explored. This is important since the first line of defence against avian influenza worldwide is surveillance. The results obtained from the infection experiments in combination with the data on bird surveillance during their migration and possible contact points with free range poultry will give a profound basis for an objective, knowledge-based risk assessment. Experiments with the different chicken lines will determine different susceptibility patterns between certain chicken breeds.

Consequently, an increase in the level of genetic resistance provides a possible means of enhancing protection of flocks against AIV infections. The analysis of the innate immune response which is important as the first line of defence against influenza viruses will increase the knowledge about crucial molecular virus-host interactions. INN-FLU Comparative structural genomics on viral enzymes involved in replication

Key words: RNA viruses, genomics, structural Acronym: VIZIER genomics, antiviral drugs, crystal structure, EC contribution: €12 905 986 Starting date: 01/11/04 bioinformatics, protein production, high- Duration: 48 months Instrument: Integrated Project throughput, screening

Summary: characterisation of emerging viruses and pre- determination. This project aims to impact the antiviral - clinical drug design. e) Target validation to assess the function design field through the identification of potential of enzymes and design strategies for virus new drug targets against RNA viruses and their inhibition. use in a comprehensive structural characterisation Aim: f) Training, implementation and dissemination. of a diverse set of viruses. The common strategies To address society’s needs, scientists need to This organisation will allow in record time, the used for the development of antiviral drugs are anticipate potential threats in order to be ready full characterisation of a viral target that can mainly based on the knowledge accumulated should they arise. The participants of the VIZIER quickly be used to design drugs, either by the through studies of virus genetics and structure. project have created a team that brings together pharmaceutical industry through the VIZIER The VIZIER project proposes to fill the existing gap the leading authorities on RNA viruses available Industrial Platform or by any R&D institution. between the necessary scientific characterisation in the EU or elsewhere as well as many leading of emerging viruses and pre-clinical drug design. European structural biologists. This team includes three partners with P4 facilities, as well as Expected results: leaders in the field of structural genomics. The VIZIER will produce an unprecedented wealth Problem: development of protocols for high-throughput of data on replicases of RNA viruses with a RNA viruses include more than 350 different (HTP) protein production means that a concerted window into the antiviral drug development. major human pathogens and most of the programme of structure determination is now A representative set of RNA-based viruses etiological agents of emerging diseases: viruses appropriate and feasible. Because they are both that belong to three major classes, profoundly of gastroenteritis (more than 1 million deaths the most likely to emerge and the most prone different in their replicative strategies, annually), measles (more than 45 million cases to genetic variability, the VIZIER consortium will will be characterised by a concerted and and 1 million deaths annually), influenza (more characterise RNA viruses that do not include a multidisciplinary effort unparalleled to date. At than 100 million cases annually), DNA stage in their replicative cycle. These virus the end of this programme, the percentage of (approximately 300 million cases annually), classes employ profoundly different replicative sequenced genomes of RNA virus species that enteroviruses and encephalitis (several million mechanisms driven by poorly characterised infect vertebrates will virtually double from cases of meningitis annually), hepatitis C virus replication machineries. Although virus-specific, 30% to 55%. As an example, the genomic (more than 150 million infected people in the the latter are the most conserved and essential characterisation of Flavivirus (ssRNA+) and world). The SARS outbreak has dramatically viral components and, thus, the most attractive Arenavirus (ssRNA-) genera, which include demonstrated how high the economic cost of an targets for antiviral therapy. a large number of human pathogens, will be epidemic caused by an emerging virus could be. systematically achieved. This negative impact is actually widening every In the framework of this project, the core day, as many governments are forced to make enzymes/proteins of the replication machinery A dramatic advancement is expected in the costly arrangements to cope with the threat carefully selected among 300 different RNA number and diversity of 3D structures of the of bio-terrorism, which lists some deadly RNA viruses, including strains of medical interest, replicative subunits, now in the one-digit range. viruses in its arsenal. will be characterised. One unique feature of VIZIER will aim at the identification of lead VIZIER, compared to other structural genomics molecules inhibiting the replicative enzymes, To meet these challenges, science needs to look projects, is the integration of major structural but will not enter into the broad field of drug for new therapeutic and prophylactic substances effort within a broad multidisciplinary study, development. Offers of cooperation will be made active against RNA viruses since those currently having virology upstream and target validation to the pharmaceutical and biotechnology industry available are scarce and of poor potency. The (candidate drug design) downstream. As a result, for further drug development, on a contractual common strategies used for the development the implementation plan of the VIZIER project is basis, and through the VIZIER Industrial Platform, of antiviral drugs are mainly based on the structured into six interacting scientific sections: which connects upfront scientific results to the knowledge accumulated through studies of virus pharmaceutical industry. genetics and structure. Yet, it is a strange paradox a) Bioinformatics, for genome annotation, target that genomic and structural characterisation of selection and data integration. Potential applications: RNA viruses was not accepted as a priority until b) Virus production and genome sequencing. With no equivalent integrated programme in very recently. The VIZIER project proposes to fill c) HTP protein production. the world, the VIZIER project will undoubtedly 68 the existing gap between the necessary scientific d) HTP crystallisation and structural have a profound impact on the field of structural challenging, innovative, and promising. promising. and innovative, challenging, is scientifically improvement to further bottlenecks putative the and target, its with together drug a finding of concept the Indeed, field. the leading by trend this reinforce undoubtedly will VIZIER, in proposed that as such section validation astrong to coupled crystallography HTP viruses. emerging future for power anticipatory the providing and process, whole the up speeding of iscapable biology structural that believed iswidely It studies. confirmatory other and toxicology of process costly and alengthy by isfollowed activity inhibitor the of discovery However,initial selected. are ‘hits’ and enzymes, purified or cells infected on screened are compounds of millions or Thousands manner. ablind in compounds screening on relies often still discovery drug current Indeed, step. design drug the to up implemented methods and concepts through considerable be to isexpected design drug on impact scientific The design. drug for targets as considered be then can proteins viral These proteins. viral of structures crystal new of bank Data Protein the in deposition the to as well as genomics) (viral viruses new of sequencing the to significantly very contribute will VIZIER that isexpected it particular, In viruses. emerging of genomics machinery. They will also identify entirely new new entirely identify also will They machinery. replication viral RNA the of understanding limited) (currently our on impact asubstantial have will products above the All leads). (drug targets protein selected for ligands or inhibitors and functions, protein assigned structures, 3D their domains, protein viral soluble sequences, genomic virus RNA are products The strategies. and technologies products, new develop will VIZIER programmes. programmes. healthcare global for priority atop - isbecoming project VIZIER the of goal ultimate -the viruses RNA against development drug Consequently, anticipation. scientific for need the and fashion atimely in drugs designing of difficulty the confirm Chikungunya) (H5N1, SARS, outbreaks viral are. Recent viruses RNA the as drugs by controlled poorly isso them of none infections, human different of fraction alarge for responsible also are parasites viral and cellular DNA-based diverse Although enterprises. industrial of development the for also but described, issues health the for only not value, strategic strong of being as isseen information such Collectively detail. of level ahigh with drugs, specific of development the for targets I S [email protected] UK Oxford Road Mansfield Oxford CEH R N [email protected] [email protected], Netherlands The Leiden 2300 9600 Box L4-36 P.O. Room E4-P, Microbiology Lumc Medical of Laboratory Dept Virology Molecular Center Medical University Leiden [email protected] [email protected], France Lyon 69338 418 Postale Case Cassin René Avenue 55 A Partners: [email protected] France Marseille Livon 13284 Charles Boulevard 58 Pharo Du Jardin Mediterranee la de Universite D Coordinator: [email protected] Italy Genova 24 16152 Perrone F Corso R N Consiglio [email protected] Germany Heidelberg 1 69117 Meyerhofstrasse Laboratory Biology E [email protected] UK 2JD OX1 Oxford Square Wellington Offices University O of University S and T [email protected] France Mundolsheim 67452 71073 b.p. Edison Thomas Rue 2 Bioxtal tu-dresden.de jacques.rohayem@mailbox. Germany Dresden 10 01069 Helmholtzstrasse D T [email protected] France Paris Roux 75724 Docteur du Rue Rage 28 La De Laboratoire I [email protected] Slovakia Bratislava 9 84506 Cesta Dubravska Zoology of Institute S [email protected] Sweden Solna 17182 Dept Virological S nfectious D nfectious nstitut Pasteur nstitut echnische Universitat Universitat echnische wedish I wedish he Chancellor, Masters Masters Chancellor, he lovak A lovak uropean Molecular Molecular uropean ciences esearch Council esearch icerche lma Consulting Group Consulting lma r Bruno Canard r Bruno resden atural E atural cholars of the the of cholars cademy of of cademy nvironment nvironment nstitute for for nstitute ationale D ationale isease Control isease xford elle elle [email protected] UK 0AX CB3 Cambridge Park Castle House Sheraton Phasing Global [email protected] Spain Madrid 28006 117 Serrano C/ S Consejo [email protected] Italy Pavia 207 27100 Abbiategrasso Via Pavia of University [email protected] France Paris 75480 Lafayette Rue 213 D le I [email protected] Germany 74 Hamburg Strasse 20359 Nocht Bernhard T for N Bernhard [email protected] Sweden Stockholm 17177 5 Nobelsvag and Biophysics Biochemistry of Dept I Karolinska [email protected] Russia Moscow Biology 119899 Chemical and of Physical Institute Belozersky N A S Moscow [email protected] Italy Cagliari Monserrato 09042 4.5 554-Km Ss Universitaria Della Citta Cagliari D Universita [email protected] Leuven 58-59 3000 Begijnhof Groot D R K ULeuven luebeck.de [email protected] Germany Lübeck 160 23538 Allee Ratzeburger Biochemie für Institut Lübeck zu Universität [email protected] Sweden Uppsala B 75105 10 Olofstgatan St University Uppsala I nstitut de R de nstitut nvestigaciones Cientificas nvestigaciones evelopment eveloppement ropical Medicine ropical uperior D uperior tate University tate ocht I ocht nstitutet echerche pour pour echerche egli S egli esearch & esearch nstitute nstitute tudi D tudi e i

VIZIER LSHG-CT-2004-511960 www.vizier-europe.org Immune response to viral respiratory infections and vaccination in the elderly

Acronym: RespViruses Key words: New respiratory viruses, immune EC contribution: €1 770 361 Starting date: 01/01/2007 response of the elderly, mouse models, diagnosis, Duration: 36 months Instrument: STREP siRNA, EGS

Summary: Problem: Expected results: During the last years some new respiratory viruses We will study the innate and acquired immune We expect to gain detailed knowledge on (HMPV/hCoVNL63/SARS/ Bocavirus/flu H5N1) response of the elderly against the new and the innate and acquired immune response to have emerged. In addition to other viruses, such known respiratory viruses. Clinical and basic emerging respiratory viruses in the elderly. as RSV, EBV and Paramyxoviruses, they are able research aspects will be addressed equally, and A thorough basic investigation of emerging to induce severe respiratory diseases in high-risk new diagnostic assays to evaluate the immune respiratory viruses will provide further handles to patients, in particular young children and the status of the elderly will be developed. Within develop antiviral strategies based on siRNA and elderly. the project an animal model for investigation external guide sequences (EGSs). Furthermore, of elderly people’s immune response will be known antivirals will be evaluated with the final established. In this model antiviral agents will goal of supporting the elderly’s immune response be tested for their ability to support the patient’s whilst reducing mortality in the elderly caused by immune response. Up to 40 000 elderly sera respiratory infections. collected during the last nine years will be tested for antibodies to emerging respiratory viruses (HCoV-NL63/HMPV/RSV/EBV/fluH5N1), providing a wide view of the epidemiology of these viruses. For a precise view of elderly people’s immune response to the viruses, a defined and uniform panel of relevant clinical data related to respiratory infections will be collected. Internationally standardised prospective data mining and a new multi-lingual software tool will be developed within the project and used by the partners.

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Coordinator Oliver Schildgen, UKB, working on virus growth in cell culture © European Commission consult any team member if necessary. if member team any consult also will and information, project acquiring for tool asoftware and system management acontact website, project the develop and create will They tools. surveillance in experience has already Hamann Mattes developed. be will disease respiratory around data medical relevant acquiring for software multilingual Therefore, data. medical of evaluation and collection and information project the of presentation and communication internal -the project the of areas two cover will Hamann Mattes SME departments, IT in consulter and developer software As access. can we that systems delivery appropriate using therapeutics antiviral potential as exploited further be can that compounds lead potent identify isto project the of goal long-term vitro in compounds these test will partners The study. the of subject the are that viruses respiratory the of sequences RNA conserved highly target that sequences guide external and siRNAs stabilised suitably synthesising and designing by project the in role acrucial plays chemistry oligonucleotide in expertise and knowledge profound its with RNA-TEC, SME Belgian The programmes). FP6 and FP5 FAIR, BIOTECH, (BRIDGE, projects European funded several in participated has and (vaccines) diseases animal of prevention of areas in active isalso INGENASA pets. or animals livestock affect that viruses for mainly field, this in history a25-year has it and sector diagnostic the for products of commercialisation and production development, research, the to dedicated company biotechnology SME is an INGENASA diseases. respiratory in development assay diagnostic for prototypes new develop to help will apartner, as expression protein recombinant and cloning acid nucleic production, antibodies monoclonal immunoassays, enzyme in skilled INGENASA, Having budget. project the of 50% approximately receive will SMEs These Spain. and Germany Belgium, in based are and RespViruses, in partners are SMEs European Three contribution: (SME) enterprise medium and Small mining. data clinical and surveillance for tool software new strategies; vaccination and treatment optimised therapies; antiviral new assays; diagnostic New P applications otential and also in suitable animal models. A models. animal suitable in also and : Tel: +49 22 82 87 11 697 11 [email protected] 82 22 +49 Tel: Germany Bonn 53105 Sigmund-Freud-Strasse 25 Virology of Dept and Parasitology Immunology Microbiology, Medical for Institute Bonn of University O Coordinator liver S liver childgen : INGENASA INGENASA Lazaro Beatriz Belgium 17-19 Leuven 3000 Minderbroedersstraat Provisorium 2 RNA-Tec S Brian Germany Nidderau 5 61130 Naumburgerstrasse Datenbanklösungen Software-und Maßgefertigte Hamann Matthias France Dijon 21079 d’Arc BP77908 Jeanne Dijon Boulevard 1 Hospital University Manoha Catherine Netherlands The Amsterdam 1105 15 Microbiology Meibergdreef Medical of Dept Virology Experimental of Laboratory Centre Amsterdam Medical Academic Hoek der van Lia Italy Siena 53100 Lotto III - Piano I - Scotte Le Policlinico Siena of University Cusi Grazia Maria Partners: Germany Aachen 30 52074 Pauwelsstrasse Aachen UK Virologie Forschungsgebiet und Aachen Lehr- Hospital University Kleines Michael Spain 39 Madrid 28037 Nobeljas Garcia nos C/H proat

RespViruses LSHM-CT-2006-037276 www.medical-surveillance.com Networking, Training, Socio-Economic and Legal Issues

This heterogeneous chapter includes projects that put influenza research into a wider context – be it the context of other related diseases, the development of networks, the link between science and policy and/or the social and legal context of the disease.

EPIZONE and VIRGIL are large Networks of Excellence with a broad-based mission to integrate Europe- wide efforts to ‘improve research on preparedness, prevention, detection and control of epizootics’ and to ‘achieve common research objectives in the area of antiviral drug resistance’, respectively. Both networks tackle highly important and topical problems and include major components of influenza: in EPIZONE as one of the most important epizootics, and in VIRGIL as a disease for which resistance to common antiviral drugs is increasingly being reported and might present a major problem in the case of a pandemic. FLU-LAB-NET is a smaller Coordination Action exclusively focusing on influenza with the goal of strengthening the network of Community Reference Laboratories and National Reference Laboratories for this disease. Improving the sharing of methodological, virological, genetic, epidemiological and clinical information on influenza will deliver a much needed research base for these laboratories with the key task of providing fast and reliable information to public health authorities, 72 policy makers and citizens in the case of outbreaks. HEALTHY POULTRY aims to serve policy makers by providing science- based decision support with regard to epizootic poultry diseases. The development of new integrated prevention and control strategies as well as their dissemination via platforms of experts, decision makers and stakeholders should help to construct this important bridge between research and the application of its result in public policy.

ConFluTech and FLUTRAIN both provide training and technology transfer in the field of avian influenza to experts in affected resource- poor countries. Since the vast majority of recent outbreaks of highly pathogenic avian influenza outbreaks were located in China and Southeast and Central Asia, capacity building in these countries is essential to control the disease at its source.

Two recently launched projects supported by the EU Public Health Programme target the social and legal context of influenza: ECORAIP aims to provide practical information and guidelines on how best to target the rural population most likely to be in direct contact with the epizootic. A key goal is to customise public health information campaigns to populations in different regions. PHLawFlu consists of two components, one of which will develop a network of interdisciplinary expertise in public health law, while the other will focus on disease preparedness specifically in the case of human pandemic influenza and prepare an inventory of European public health laws pertaining to this problem. One important goal will be to detect inconsistencies and regulations that do not take into account modern public health evidence.

This last chapter of the present catalogue thus provides examples of the connections and interdependencies between influenza research and a number of other areas. Related scientific fields such as other infectious diseases or more remote disciplines like social sciences or law frequently make important contributions to the implementation of research results, be it through public policy, training and technology transfer or awareness raising and information campaigns. Development and enhancement of laboratory networks for avian influenza

Acronym: FLU-LAB-NET EC contribution: €930 000 Starting date: 01/05/2007 Key words: Avian influenza, laboratory, network, Duration: 36 months Instrument: Coordination Action FLU-LAB-NET

Summary: Aim: Potential applications: FLU-LAB-NET provides new opportunities To share and exchange methodological, By reinforcing and enhancing the existing for enhancement and reinforcement of the virological, genetic, epidemiological and clinical Community Reference Laboratory and National Community Reference Laboratory and National information on influenza. The network will Reference Laboratories network for avian Reference Laboratory network for avian present up-to-date, quality information on influenza within the European Union, FLU- influenza (AI) within the EU. This will strengthen influenza activities for scientists, policy makers, LAB-NET will facilitate improvements and harmonisation and development of laboratory professionals and the public. It will also encourage harmonisation of laboratory and diagnostic and diagnostic methods, coordination of research the identification of duplicate areas of work methods. Following the development and efforts and sharing of expertise. Rapid responses including surveillance and research projects at a implementation of FLU-LAB-NET as a web-based, to national and global emergencies with data European level. global interactive community, the EU Member sharing will be key areas of exploitation, State, third country and INCO partner laboratories contributing to a European laboratory task force will have facilities to optimise rapid, formal capability for AI in animal species. Rapid, formal Expected results: interactive communications forums. This will in interactive communications will be addressed a) Enhancement and reinforcement of the turn be extended to participating laboratories through web-based forums. existing Community Reference Laboratory and involved in influenza research in domestic National Reference Laboratory network for mammals, and provide a hub to allow fostering of Laboratories involved in influenza research avian influenza within the European Union formal links and coordination with corresponding on domestic mammals will also participate. Member States. human, swine and equine influenza networks. FLU-LAB-NET will also foster formal links and b) Development and implementation of coordinate with corresponding human, swine web-based, global interactive communities In addition, FLU-LAB-NET will allow for the and equine influenza networks. FLU-LAB-NET facilitating rapid, formal interactive coordination of research efforts and data provides opportunities for identification and communications forums. exchange, as well as development and sharing development of the complementarities of global, c) Strengthening harmonisation and development of expertise. Rapid responses to national and multi-disciplinary influenza research programmes. of laboratory and diagnostic methods, global emergencies, with data sharing, will also Strategically important third country and INCO coordination of research efforts and sharing of be a key area of exploitation, contributing to partners are also included in this network, in expertise. a European laboratory task force capability for order to raise laboratory standards and benefit d) Facility for rapid responses to national and AI in animal species. FLU-LAB-NET provides from knowledge sharing. This will promote global emergencies, with data sharing. opportunities for identification and development greater trust, understanding and early access to e) Extension of knowledge sharing and of the complementarities of both EU and global, information that may be of importance to both laboratory support to strategically important multi-disciplinary influenza research programmes. veterinary and public health in the EU. third country and INCO partner laboratories. This will promote greater trust, understanding f) Participation of laboratories involved in and early access to information that may be of influenza research in domestic mammals. importance to both veterinary and public health Problem: g) Fostering of formal links and coordination in the EU. The current global H5N1 crisis has resulted in with corresponding human, swine and equine a significantly increased demand for laboratory influenza networks. capacity and capabilities for AI diagnosis and surveillance. The rapidly expanding growth in AI work has highlighted the benefits of closer collaboration within existing AI laboratory networks, which we propose to address. The spread to humans has also highlighted real concerns of the pandemic potential of H5N1, reinforcing the need for closer integration between veterinary and public health laboratories. 74 eu .

Coordinator: Dr Veronique Jestin Dr Eveline Wodak David Graham Dr Yatinder Singh Binepal

Dr Ian Brown Agence Française de Sécurité AGES IVET Moedling Agri-food and Biosciences Kenya Agricultural Research net

Veterinary Laboratories Agency Sanitaire des Aliments Robert Koch Gasse 17 Institute Institute - - Weybridge AFSSA - site de Ploufragan Moedling Veterinary Sciences Biotechnology Center Woodham Lane B P 53 Austria Division P.O. Box 57811

New Haw 22440 Ploufragan [email protected] Stoney Road 00200 Nairobi lab Addlestone KT15 3NB France Dr Zenon Minta Stormont Kenya - Surrey [email protected] National Veterinary Research Belfast BT4 3SD [email protected]; UK Pat Raleigh Institute Northern Ireland [email protected] flu

Tel: +44 19 32 35 73 39 Central Veterinary Research Dept of Poultry Diseases UK Emina Rešidbegović . [email protected] Laboratory Al Partyzantów 57 [email protected] Veterinary Faculty Poultry Dept of Agriculture and Food 24100 Pulawy Christine Monceyron Centre Partners: Laboratories Poland Jonassen Zmaja od Bosne 90 Dr Thierry van den Berg Backweston Campus [email protected] National Veterinary Institute 71000 Sarajevo

Veterinary & Agrochemical Staccumny Lane Dr Miguel A S P Torres Section for Virology and Serology Bosnia-Herzegovina www Research Centre Celbridge Fevereiro P.O. Box 8156 Dep [email protected] Avian Virology & Immunology Co Kildare Laboratorio Nacional de 0033 Oslo Prof. Chukwudozie Daniel Unit Ireland Investigacao Veterinarial Norway Ezeokoli Dept of Small Stock Diseases [email protected] Estrada de Benfica 701 christine.monceyron-jonassen@ The University of the West Groeselenberg 99 Ilaria Capua Lisboa vetinst.no Indies 1180 Brussels Istituto Zooprofilattico Portugal Dr Gabriela Goujgoulova School of Veterinary Medicine Belgium Sperimentale delle Venezie [email protected] National Diagnostic Research Faculty of Medical Sciences [email protected] Laboratorio di Virologia agricultura.pt Veterinary Medical Institute Clinics Laboratory Poul Henrik Jørgensen Viale dell’Università 10 Dr Olga Zorman Rojs 15 Pencho Slaveikov Blvd St Augustine Debra Elton Danish Institute for Food and 35020 Legnaro University of Ljubljana 1606 Sofia Trinidad and Tobago Animal Health Trust Veterinary Research Italy Veterinary Faculty NRL of AI and NDV West Indies Lanwades Park Hangoevej 2 [email protected] Institute of Poultry Health and 190 Lomsko shose Blvd [email protected] Kentford Aarhus N Dr Kyriacos Georgiou Protection 1231 Sofia [email protected] Newmarket CB8 7UU Denmark Veterinary Services Gerbičeva 60 Bulgaria Prof. Hualan Chen Suffolk [email protected] 1417 Athalassa 1000 Ljubljana [email protected] Chinese Academy of UK Dr Martin Beer Nicosia Slovenia Dr Aurelia Ionescu Agricultural Sciences [email protected] Friedrich-Loeffler-Institute Cyprus [email protected] Institute for Diagnosis and Harbin Veterinary Research Kristen Van Reeth Federal Research Institute for [email protected] Miroslav Mojžiš Animal Health Institute University of Gent Animal Health Mrs Ieva Rodze State Veterinary Institute Bucharest Animal Influenza Laboratory of Sint-Pietersnieuwstraat 25 FLI Insel Riems National Diagnostic Centre of Zvolen Romania Ministry of Agriculture Gent Boddenblick 5a Food and Veterinary Service National Reference Laboratory [email protected] 427 Maduan St Belgium 17493 Greifswald-Insel Riems Lejupes Str 3 for AI and ND [email protected] 150001 Harbin [email protected] Germany Riga Poddrahami 918 Prof. Richard Hoop China William John Pagett [email protected] Latvia 96086 Zvolen Institute of Veterinary Bacteriology [email protected] Nederlands Instituut Mr Ants Jauram [email protected] Slovakia National Reference Centre for Dr Khalid Naeem voor onderzoek in de Estonian Veterinary and Food Prof. Vilmos Palfi [email protected] Poultry Diseases National Agricultural Research Gezondheidszorg Laboratory Central Veterinary Institute Liisa Sihvonen Dept of Poultry Diseases Centre European Influenza Kreutzwaldi 30 Tabornok Utca 2 Finnish Food Safety Authority Winterthurerstr 270 National Reference Laboratory Surveillance Scheme Tartu Budapest Evira 8057 Zürich for Poultry Diseases Otterstraat 118-124 Estonia Hungary Virology Unit Switzerland Park Road Utrecht [email protected] [email protected] Mustialankatu 3 [email protected] 45500 Islamabad The Netherlands Vasiliki Rousi Dr Ben Peeters 00790 Helsinki Dr Wlodek Stanislawek Pakistan [email protected] Ministry of Rural Development Central Institute for Animal Finland Investigational and Diagnostic [email protected] Concepción Gómez-Tejedor and Food Disease Control, Division of [email protected] Centre-Wallaceville Vladimir Savić Laboratorio Central de Centre of Athens Veterinary Virology Prof. Sándor Belák Biosecurity New Zealand Croatian Veterinary Institute Veterinaria (Algete) Institutions P.O. Box 2004 The National Veterinary Institute Ministry of Agriculture and Forestry Poultry Centre Ctra De Algete, km. 8 25 Neapoleos St Houtribweg 39 Ulls vag 2B P.O. Box 40742 Heinzelova 55 28110 Algete Athens 8203 AA Lelystad Uppsala 5018 Upper Hutt 10000 Zagreb Madrid Greece The Netherlands Sweden New Zealand Croatia Spain SSPE-CT-2007-044453 [email protected] [email protected] [email protected] [email protected] [email protected] [email protected] FLU-LAB-NET Training and technology transfer of avian influenza diagnostics and disease management skills

Acronym: FLUTRAIN EC contribution: €1 809 133 Starting date: 01/03/2007 Key words: Avian influenza, training, diagnostic Duration: 36 months Instrument: Coordination Action tools, exchange

Summary: Problem: Aim: The present worldwide avian influenza (AI) crisis Avian influenza infections have become of Avian influenza infections caused by several has highlighted the need for comprehensive increasing relevance both from the animal and subtypes are endemic in vast areas of the world, training and the transfer of technology to EU human health perspectives. With the extension particularly in developing countries and countries accession and international cooperation target of the H5N1 epidemic from Asia to Eastern where poverty is widespread. Under these (INCO) countries with the clear goal of aiding Europe and Africa, notwithstanding the efforts circumstances AI infections are very difficult to these countries in combating AI with the most of international organisations, there is clear control, due to the lack of funds to train staff, up-to-date diagnostic and disease management evidence that in the current situation the attempts produce or purchase diagnostic reagents or apply procedures. to stop the infection’s progress are insufficient. modern diagnostic technology. The objective There are several factors that are contributing to of FLUTRAIN is to bridge the gap of knowledge The FLUTRAIN project will first approach the the spread, an important one being the delay in between EU scientists and colleagues in AI need for training by providing two workshops identifying primary outbreaks in a given area in affected countries. This project has the objective over the duration of the project that will call on a timely manner. This has highlighted the need of supporting countries affected by AI infections experts in the AI field to pass on their expertise for comprehensive training and the transfer of through training and the transfer of knowledge in the diagnosis and management of AI to technology to accession and INCO countries and technology. participants from accession and INCO countries. with the clear goal of aiding these countries Training opportunities will also be provided in in combating AI using the most up-to-date partner laboratories in order to consolidate the diagnostic and disease management procedures. information and practical experience gained during the workshops. In addition, a CD-ROM will be prepared containing essential information provided during the workshops. A website will be developed which will enable participants and the general public to access the training programmes and will include on-line discussion forums between trainees and trainers.

Another goal of FLUTRAIN will be the transfer of technology to accession and INCO countries. This will include the provision of new, simplified and cost-effective diagnostic methods and reagents. It will also involve the transfer of deliverables, both for serological and virological diagnosis, that have been (or will be) developed in three European projects - AVIFLU, Lab-on-Site and FLUAID. Finally, FLUTRAIN will identify and supply funds, in a bilateral manner, for one-off specific support missions that will target specific AI problems in recipient countries.

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Training in avian influenza techniques at the FLUTRAIN © coordinator’s institute (IZSVe, Italy –2006) Ex e) Generation of an interactive website. website. interactive an of Generation e) the containing aCD-ROM of Generation d) can that trainers of agroup of Generation c) and general addressing Twob) workshops cost molecular, and classical of Generation a) ELISA) that can be exported to accession and and accession to exported be can that ELISA) PCR, (reagents, tools diagnostic effective distributed to trainees and partners. and trainees to distributed be will that workshops the of material diagnosticians. laboratory scientists\ other to on knowledge their pass AI. to related topics specific INCO countries. results pected : Tel. +39 04 98 08 43 71 43 [email protected] 08 98 04 +39 Tel. Italy Venezie Legnaro delle Sperimentale Zooprofilattico Istituto Disease Newcastle and Influenza Avian for Reference Laboratory National and OIE\FAO D Coordinator: r I laria Capua Capua laria Italy Italy Brescia Lombardia dell’Emilia-Romagna e della Sperimentale Zooprofilattico Istituto D [email protected] Sweden Sciences of Agricultural University Swedish D [email protected] Belgium Medicine Merelbeke Veterinary of Faculty University Ghent D [email protected] Belgium Brussels Center Research Agrochemical and Veterinary D [email protected]/ Netherlands The Lelystad (CIDC-Lelystad) Health Science and Animal for Institute D [email protected] UK Surrey Addlestone Lane Woodham Dept Agency Virology Laboratories Veterinary D Partners: [email protected] Italy Padua of University Health Public of Dept D [email protected] Denmark Research and Food for Veterinary Institute Danish D [email protected] r Maura Ferrari r Maura r S R van r Kristien r T Koch r Guus Banks r Jill r A Henrik r Poul hierry van den Berg den van hierry andor Belak andor lessandra Piccirillo Piccirillo lessandra eeth [email protected] Sweden AB Uppsala Biotech Svanova D [email protected] Ireland Ltd Carlow Productions Capture Mc Kevin Mr r Malik Merza r Malik D ermott ermott A National Center for Veterinary Veterinary Diagnosis for Center National D [email protected] Nigeria Vom Veterinary Institute Research National D [email protected] Turkey Medicine Veterinary University Firat D [email protected] Slovenia Ljubljana Faculty Veterinary Ljubljana of University D [email protected] Croatia Institute Zagreb Veterinary Croatian D [email protected] Romania Bucharest Health and Animal Diagnosis for Institute D [email protected] USA Iowa Research Laboratory Poultry Agriculture of Southeast Department US D [email protected] Bulgaria Institute Medical Veterinary Research and Diagnostic National D [email protected] Vietnam Hanoi Dong-da St Giai-Phong Lane 11-78th Health Animal of Dept sso r T r T r A r O S r Vladimir r I r D r Georgi Georgiev Georgiev r Georgi onescu A onescu ony Joannis Joannis ony hanh Long T Long hanh ykut O ykut ennis S ennis lga Zorman- lga c iated p iated zdarendeli enne urelia urelia avic avic artners: o R ojs

FLUTRAIN SSPE-CT-2006-044212 www.flutrain.eu Capacity building for the control of avian influenza through technology transfer and training

Acronym: ConFluTech EC contribution: €547 255 Starting date: 01/07/07 Key words: Avian influenza, polymerase chain Duration: 36 months Instrument: SSA reaction, epidemiology, disease outbreak

Summary: Problem: Aim: Avian influenza (AI) or ‘bird flu’ is a highly Avian influenza is a highly contagious viral The overall aim of this proposal is to facilitate contagious viral infection which can affect all infection which can affect all species of birds and technology transfer and training to promote species of birds and can manifest itself in different can manifest itself in different ways depending capacity building in INCO (International ways depending mainly on the pathogenicity of mainly on the pathogenicity of the virus Cooperation) target countries, with a particular the virus involved and on the species affected. involved and on the species affected. The highly emphasis on countries that border the EU, for The highly pathogenic avian influenza (HPAI) pathogenic avian influenza (HPAI) virus causes better control of avian influenza and general virus causes serious disease with high mortality serious disease with high mortality (up to 100%). outbreaks of infectious diseases in livestock. (up to 100%) - notifiable to the OIE. Worryingly, Worryingly, HPAI has been shown to infect and This will be achieved through the organisation of HPAI has been shown to infect and cause death cause death in humans in a number of countries technical workshops and training courses in the in humans. Up to now, a total of 103 deaths have such as Vietnam, Turkey and Iraq. Besides the fact following fields: been recorded due to HPAI infection in a number that a number of countries were surprised by the of countries such as Vietnam, Turkey and Iraq outbreaks, an even greater number of developing a) Molecular diagnostic tools for pathogen (WHO, 21 March 2006). countries do not have adequate tools to detect detection and differentiation. and differentiate HPAI and are lacking experience b) Standardisation and validation of diagnostic Beside the fact that a number of countries were to manage the outbreak of the disease. Thus there tools according to OIE instruction. surprised by the outbreaks, an even greater is an urgent need for technology transfer and c) Epidemiological tools for monitoring and number of developing countries do not have training. modelling avian influenza outbreaks. adequate tools to detect and differentiate HPAI d) Management of disease outbreaks. and are lacking experience to manage the outbreak of the disease. Thus, there is an urgent need for technology transfer and training. To fulfil these gaps, the partners of this consortium will:

a) organise technical workshops to facilitate technology transfer particularly in the field of molecular diagnostic tools for pathogen detection and differentiation, to reinforce epidemiological analysis for monitoring and modelling of avian influenza especially and to respond to outbreaks of infectious diseases of livestock in general;

b) provide training through organisation of seminars and short-term courses in well qualified laboratories of a number of EU Member States;

c) organise technical workshops, courses and training in the INCO target countries to improve the technical experimental level of the staff and laboratories in charge of livestock infectious diseases.

78 de . Expected results: Potential applications: Partners: The project will result in the transfer of The following technical and scientific advances Prof. Sándor Belák Iran technologies required for specific and rapid are expected: Swedish University of Tel: +98 21 66 44 69 diagnosis of diseases, disease surveillance and a) Transfer of knowledge and technology for Agricultural Sciences [email protected] Ulls väg 2B Dr Lokman Taib Omer management of disease outbreak. The impact of surveillance systems to exploit and understand borstel 75189 Uppsala Dohuk University - the project in the introduction of techniques disease transmission pathways, establishment Sweden Dohuk Research Center Tel: +46 18 67 41 35 Faculty of Veterinary Medicine fz . required for the implementation of the required of reliable indicators and tools for disease [email protected] Dohuk control measures. In addition, the project will monitoring activities (analytical and modelling) Prof. J M Sánchez-Vizcaíno Iraq Catedrático de Sanidad Animal Tel: +96 47 50 45 04 789 serve as a platform for technology transfer for to describe the present situation and predict Universidad Complutense [email protected] the improvement of livestock disease control the future course of AI as well as for a better Dpto Sanidad Animal Prof. Dr Darem Tabbaa www measures and will contribute to reducing the response to diseases. Avda Puerta de Hierro s/n Faculty of Veterinary Medicine 28040 Madrid Al-Baath University risk of the spread of transboundary diseases to b) Standardised diagnostic tools and methods. Spain Hama Europe. The objectives of the project contribute to c) Tools in decision making, communication and Tel: +34 91 39 44 082 Syria [email protected] Tel: +96 33 35 12 640 the realisation of EU policy regarding food security information systems. Dr Viorel Alexandrescu [email protected] and food safety in the developing countries. Cantacuzino Institute Prof. Dr Labib Sharif Romania Faculty of Agriculture and Moreover, they contribute to reducing the risk of Tel: +40 21 31 84 410 Veterinary Medicine diseases spreading to Europe and to protecting [email protected] The Jordan University of Europe’s livestock. Dr Iyisan, Ayse Selma Science and Technology Pendik Veterinary Control Jordan Cad No 10 Tel: +96 22 72 01 000 81480 Pendik sharifqjust.edu.jo Ankara Istanbul Prof. Rogdakis Emmanouil Turkey Production – Agricultural Tel: +90 21 63 90 12 80 University of Athens [email protected] Laboratory of General and Dr Hokobyan Hohannes Special Animal Technology State Veterinary Inspection Greece Armenia Tel: +30 21 05 29 44 31 Tel: +37 41 09 53 841 [email protected] [email protected] Ass. Prof. Dr Rositsa Kotseva Prof. Jafarov Mamedtagi National Reference Laboratory Ganja State Agricultural of Influenza and Acute Academy Respiratory Diseases Azerbaijan 26 Yanko Sakazov Blvd Tel: +99 42 25 61 731 Sofia [email protected] Bulgaria Dr Giorgi Meskhishvili Tel: +35 92 83 10 030 Georgian State Zootechnical [email protected] Veterinary Academy Dr Adama Diallo Coordinator: Tbilisi Animal Production and Health Section Jabbar Ahmed Georgia FAO/IAEA Joint Division Research Center Borstel Tel: +99 59 92 67 278 Wagramer Strasse 5 Parkalle 22 [email protected] P.O. Box 100 23845 Borstel Ass. Prof. Prviz Shayan 1400 Vienna Germany University of Tehran Austria Tel: +49 45 37 18 84 28 Faculty of Veterinary Medicine Tel: +43 12 60 02 60 52 [email protected] Dept of Parasitology [email protected] SSPE-CT-2006-044462 ech T lu F on C European vigilance network for the management of antiviral drug resistance

Acronym: VIRGIL EC contribution: €9 000 000 Starting date: 01/05/2004 Key words: Antiviral, drug, resistance, hepatitis, Duration: 48 months and beyond Instrument: Network of Excellence influenza, flu, HCV, HBV, virus

Summary: Problem: all aspects of antiviral drug resistance, regardless VIRGIL (for Vigilance against Viral Resistance) is The development of new antiviral therapies over of the type of virus concerned (surveillance, the first European surveillance network capable of recent decades constitutes major progress in the diagnosis, basic virology and modelling, host- addressing current and emerging antiviral drugs treatment of viral infections with a considerable inherent factors, pharmacology, technological resistance developments in the field of influenza impact on life expectancy and quality for innovation, socio-economic impact). It currently and viral hepatitis. Coordinated by Inserm (the patients. In the case of highly contagious viruses comprises more than 70 laboratories, including French Institute for Health and Medical Research) such as influenza, antiviral drugs can also be six biotech small and medium enterprises (SMEs), and supported by a grant from the Priority 1 Life used to control epidemics, and, in the case in more than 16 Member States of the European Sciences, Genomics and Biotechnology for Health of the emergence of a pandemic virus could Union and beyond. programme in the Sixth Framework Programme even contribute to slowing or preventing its of the EU, the network’s activities started in propagation. The other goals of VIRGIL are to: May 2004 with the initial task of integrating a) provide the pharmaceutical industry with the fragmented European capacities and major One of the consequences, however, of this centralised management and standardised expertise in the field into a single coherent success is the high frequency of drug resistance, computer tools to conduct large-scale clinical Network of Excellence. VIRGIL initially gathers 60 which may be due to the patient (host), or to the trials on viral hepatitis and influenza; organisations, including more than 60 academic virus, or to a combination of both. In particular, b) rationalise the use of antiviral drugs for the laboratories and seven companies from 14 the emergence of resistant viral strains during benefit of patients and health systems and to European countries and beyond. treatment (Darwinian selection), in the absence contribute to the development of new more of adequate follow-up, can lead to treatment effective and less expensive drugs; VIRGIL is structured into seven integrated failure, or even epidemic diffusion of the resistant c) transpose these concepts of excellence to other platforms centred on the patients with each strains. It has been estimated, for example, that regions of the world, including developing focusing on a topic contributing towards the after four years of hepatitis B antiviral therapy countries where the viruses studied are project objectives, from surveillance of resistant with lamivudine, 70% of patients present endemic; viruses in Europe, innovation in diagnostic tools, resistances. Rational and logical use of antiviral d) constitute the basis for a future European analysis of the virological mechanisms of antiviral drugs is essential to minimise the clinical and Virology Foundation. treatment efficacy and resistance development epidemiological impact of resistance. Treatment and pharmacology to studies on host (patient- strategies must therefore be adapted in real time In the first phase, three disease models were related) factors and the socio-economic impact to changes in viral populations. This monitoring selected (influenza, hepatitis B and hepatitis C) of antiviral drug resistance. VIRGIL benefits requires a multidisciplinary approach, as it for the development and testing of working tools. from a central management unit that offers is based on epidemiology, clinical follow-up, Chronic hepatitis (15 million people infected industrial partners a single contact point as well diagnostics and basic research and involves public in Europe) is responsible for two thirds of all as integrated services for conducting large-scale institutions as well as private laboratories. cases of cirrhosis and liver cancer, and 50% of European clinical trials. Finally, the network patients present drug resistances. Influenza, with is also committed to training students and its annual epidemic waves, is responsible for researchers within and outside of the participating Aim: 21 000 deaths per year in France and 114 000 organisations and to spreading VIRGIL’s practices The resources and skills allowing control of the hospitalisations. The network is also reactive of excellence and the resulting knowledge various aspects of resistance already exist in and can respond to emergency situations such as throughout the medical community and the European private and public laboratories, but influenza pandemics. general population. until recently they were fragmented, constituting a major obstacle to progress of knowledge. The primary goal of VIRGIL is to gradually integrate Results: resources and skills dispersed throughout Europe Less than three years after the creation of VIRGIL, to achieve common research objectives, including the preliminary results obtained on antiviral the study of the socio-economic dimension of drugs used to treat influenza and hepatitis B and antiviral drug resistance. VIRGIL was therefore C demonstrate the pioneer role in Europe of an designed to be a virtual institute organised into integrated approach linking basic research and 80 seven collaborative platforms able to deal with clinical research. they are used rationally, and if the emergence of of emergence the if and rationally, used are they if useful be only will drugs These oseltamivir. of stocks up set have EU the of countries all time, present the At WHO. the to according high, remains still pandemic a of emergence of risk the and birds, domestic and wild in infections epizootic numerous inducing Africa, and Asia in circulate to continues virus H5N1 The pandemic. influenza an of event the in mobilised be could programme this of context the in developed infrastructures and skills the influenza, seasonal on benefit immediate their from Apart territory. its in circulating strains resistant monitor to skills necessary the develop autonomously to country European each allow to is objective The UK). London, Agency, Protection (Health Zambon Maria Dr by directed team VIRGIL the by up set been also has centres reference influenza national WHO of directors for tests resistance NI on programme training A developed‘. be to needs resistance NI on knowledge when time the at ‘just arrives initiative this him, to According ).‘ (NIs Inhibitors Neuraminidase to resistance on knowledge of diffusion and study surveillance, real-time the in akey role plays VIRGIL influenza, of aspects various on working people of efforts the integrating ‘By says: UK), (London, centres reference influenza world WHO four the of one of Hay, Alan Prof. director impact. epidemiological their and property) infectious (symptoms, features clinical associated their morphological), (genetics, strains resistance of characteristics the of analysis combined allow will approach VIRGIL The patients). immunosuppressed (children, population the of subgroups susceptible in viruses, influenza drug-resistant of emergence of conditions the seasons, consecutive several over study, to laboratories reference influenza national of network WHO the and (EISS), Scheme Surveillance Influenza European the with acollaboration initiated has VIRGIL vitro demonstrated been have cross-resistance of absence and synergy whose molecules these of combinations using trials clinical of design the allow will This treatments. for targets viral new as well as interferons, effective more new molecules, antiviral between antagonisms and synergies identified have studies several C, hepatitis of case the In guidelines. official of establishment the for authorities health by adopted be could results these Liver), the of Study the for Association (European EASL as such societies scientific and VIRGIL between links the of result a As B. hepatitis of treatment the for strains) resistant multi and entecavir (adefovir, drugs antiviral marketed newly to resistances characterise precisely to first the were teams VIRGIL . in vitro in

in in established a number of standardised criteria for for criteria standardised of anumber established has VIRGIL existence, of years two first its During trials clinical large-scale of conduct the for place important an Europe Making network. dense sufficiently a to according EU the throughout up set surveillance continuous by controlled is strains viral resistant represented by VIRGIL’s partners. According According VIRGIL’s by partners. represented excellence of regions various with them linking by tissue economic European the in (biotechs) SMEs of integration the promote and groups pharmaceutical major with services trial clinical centralised develop to plans VIRGIL ribavirin. and peg-interferon with therapy dual to HCV of resistances and tenofovir, or entecavir as such molecules new to HBV of resistances characterise to VIRGIL by databases these on initiated been have trials clinical Several allowed: has process standardisation This trials. various in obtained results the of comparison allow will VIRGIL of members all by criteria these of adoption as step, isacrucial This trials. clinical in collection data in France therefore constitutes a major advantage. amajor constitutes therefore France in pole competitiveness Rhône-Alpes the in VIRGIL of SMEs‘. of Installation growth sustainable the for condition essential isan market international ‘access the to Paris), Transfert, (INSERM VIRGIL of coordinator logistic Weinbach, Jerome Dr to c) the design and development of on-line on-line of development and design the c) VIRGIL of members all by adoption the b) to form consent informed an of elaboration the a) the questions raised by particular studies. particular by raised questions the of afunction as interest of patients of selection allow also will They etc.). resistance secondary or primary follow-up, treatment, of composition (profile, patients on information of exchange secure and standardised allowing databases values; cut-off load viral on based drugs C antiviral Band hepatitis to resistance drug secondary and primary of definitions standardised of EU legislation; current of requirements ethical the with complies strictly research this that ensure to designed was form this site, intranet VIRGIL’s from downloadable and 10 languages Translated into Cpatients. Bor hepatitis from taken samples biological on research perform

VIRGIL LSHM-CT-2004-503359 www.virgil-net.org 82 transfert.fr 139 jerome.weinbach@inserm- 30 50 15 +33 Tel: France Paris 75013 Watt Rue 7 Transfert Inserm D Manager: Project 971 [email protected] 81 26 47 +33 Tel: France Lyon Thomas 69003 Albert France de Cours 151 Universitaire Institut Liver and Dept. 271 Unit INSERM Zoulim Fabien Prof. S Coordinator: cientific Coordinator: cientific r Jerome Weinbach r Jerome Partners: Belgium Belgium Paediatrics of Dept de Louvain Catholique Université E Poland Biotechnology of Dept Gdansk of University Bielawski Krysztof Switzerland Genève de Université N Francesco UK Birmingham of University D Greece Hospital General Papageorgiou Germanidis Georgios Spain de Provincial Clinic Hospital Bruix /Jordi Forns Xavier Spain Hebron Valle Universitario Hospital Buti Maria R Italy Molecular of Medicine Institute Venetian A Netherlands The Centre Medical Erasmus S France des d’Observation Réseaux Cohen Jean-Marie Ltd UK Virology Retroscreen O John Netherlands The Medical Center University Leiden S Willy Leuven Belgium Universiteit Katholieke N Johan Germany Düsseldorf Universitätsklinikum Bode Johannes Germany Munchen Maximilians-Universat Ludwig Mueller D /Helmut Pape Gerd UK Council Research Medical British A Germany Heidelberg Universitätsklinikum R UK Laboratory Health Public Agency Central Protection Health Cane /Pat Zambon Maria France XII-Val-de- Marne Paris Université Pawlotsky Jean-Michel Germany Hannover Hochschule Medizinische Manns Michael France 1 Bernard Lyon Claude Universite Lina Bruno Médicale France et Santé Recherche la la de de National Institut T /Christian Zoulim Fabien Barcelona Rotterdam Epidémies des et Maladies repo / Christian Brechot /Christian repo tienne S tienne S olko afaele E afaele Bartenschlager alf lfredo A lfredo Hay lan avid Mutimer avid iepolder /T iepolder xford paan chalm eyts steban Mur / Mur steban okal lberti egro homas homas Universität des Saarlandes des Universität S Medicine and Science, of Technology College Imperial Karayiannis T Howard III Spain Carlos AEIS-Hospital S Vincente Gilbert D Gilbert S Germany Hamburg Universitätsklinikum Petersen Joerg Italy Del Raffaele San Centro Fondazione Guidotti Luca Sweden University Uppsala D Helena Freiburg Germany Universitätsklinikum D N /Michael Baumert /T Blum Hubert France I Fourier Grenoble Joseph Université R Cientificas Spain de Investigaciones Superior Consejo O Juna Marburg Germany Philipps-Universität Garten Wolfgang D Hans Animals Germany of for Diseases Centre Virus Research Federal O France Pasteur Institut S Sanità di Italy Superiore Istituto I Germany Italy Bari of University S /T Pastore Guiseppe Ltd UK Pharmaceuticals Riotech T Mark Sweden Weiland Ola Institute Karolinska S Matti Germany Berlin Virchow-Klinikum Campus Charité T Parma di Italy Ospedaliera Azienda Missale /Gabriele Ferrari Carlo Germany Koch-Institute Robert T Germany Giessen Justus-Liebig-Universitaet- S UK of Oxford University the of and Scholars Masters Chancellor, Klenerman N France Scientifique la de Recherche National Centre UK Eppendorf Tabor Monte Universitätsklinikum sabella D sabella tefan Zeuzem tefan téphane Bressanelli / Bressanelli téphane tephan Pleschka tephan ylvie van der Werf der van ylvie homas Berg homas Wolff horsten antantonio obertus R obertus arius Moradpour arius icole Zitzmann / Paul /Paul Zitzmann icole liver Planz liver rtin hursz ieter Klenk / Klenk ieter aellberg anielson eléage homas / Peter / Peter homas onatelli oriano uigrok homas homas eresa eresa asal / asal Germany Aachen of University T Christian Luxembourg Public- Recherche de Centre S Jean-Claude UK Westfield College and Mary Queen Foster Graham Netherlands The MedlawConsult E Muenster of University S Cesalpino Italy Andrea Fondazione Levrero Massimo Netherlands The for Institute Netherlands Paget John UK of Cambridge University the of and Scholars Masters Chancellor, S /D Grenfell Bryan Israel University Bar-Ilan A France SA bioMérieux Baccala /Glaucia Vernet Guy SA France Pharma BioAlliance A Gilles Austria Innsbruck of University Puerstinger Gerhard Sweden AB Tripep A Switzerland Bern of University Cerny R Juerg Luxembourg du Santé Germany Research Services Health tephan Ludwig tephan vert-Ben Van Veen Van vert-Ben mith vidan N vidan nders Vahlne nders eichen /A eichen venard eumann rautwein chmit ndreas ndreas erek erek Tel: +35 72 28 92 885 [email protected] 92 28 72 +35 Tel: Cyprus Nicosia 1678 20537 Box P.O. St Kallipoleos 75 Cyprus of Sciences University Biological of Dept GKostrikis Leondios Prof. 82 25 [email protected] 35 26 21 +90 Tel: Turkey Istanbul 34390 Çapa Virology Laboratory Dept Medicine of Microbiology Faculty Istanbul I S Prof. [email protected] 646 49 97 95 74 Tel: Russia St Moscow 111123 Novogireyevskaya 3a of Institute Epidemiology Research Central Diagnostics Molecular for Center Chulanov Vladimir Prof. asso stanbul Üniversitesi Üniversitesi stanbul c elim Badur elim iate m iate e mb ers:

VIRGIL LSHM-CT-2004-503359 www.virgil-net.org Development of new integrated strategies for prevention, control and monitoring of epizootic poultry diseases Acronym: HEALTHY POULTRY Key words: Avian influenza, epizootic poultry EC contribution: €1 119 404 Starting date: 01/11/2004 diseases, disease control and prevention, policy Duration: 36 months Instrument: STREP and decision making

Summary: Problem: Aim: When highly pathogenic strains of influenza The European Union aims at assuring a high level The primary aim of the project is to provide break out in poultry, the consequences can be of animal health and animal welfare without scientifically-based support to decision makers in devastating (around 30 million chickens had to be compromising the functioning of the internal the field of epizootic poultry disease prevention killed during the avian influenza (AI) outbreak in market. Nevertheless, in the last decade, several and control. The Netherlands in 2003). Destruction of infected epizootics of AI occurred throughout the EU. The objectives of the project are: birds is the primary means of control today but, These had a devastating veterinary and economic a) to develop new integrated strategies for as the Dutch crisis proves, even such drastic impact. Moreover, fear amongst the population prevention, control and monitoring of epizootic measures cannot totally avoid disaster. Given increased because of a possible impact on human poultry diseases; the global spread of the H5N1 AI subtype, policy health as well, particularly during the last couple b) to analyse these strategies in a comprehensive makers across Europe are looking at revising their of years. Finally, control of AI currently coincides way; strategies to better prepare for future outbreaks with severe problems related to socio-ethical c) to provide guidelines for the implementation of of epizootic diseases in poultry. They want to issues and animal welfare. these strategies in EU Member States; know how best to avoid infection and, when d) to develop user friendly toolboxes for strategy infections do occur, how best to limit their spread Intensive trade contacts (of animals and evaluation; and impact. poultry products) between Member States pose e) to disseminate project results to a broad considerable risks to poultry in the EU once a relevant audience. HEALTHY POULTRY brings together seven single Member State is struck by AI. academic institutions from Italy, the Netherlands, Quite obviously, strategies and measures for Germany and Hungary to assess current scientific prevention and control of AI need improvement understanding of epizootic AI. Researchers to fulfil the EU objectives. Future prevention and will then use this scientific basis to suggest control of AI should be more efficient, ethically new strategies for their prevention, control acceptable and less costly. Self evidently, because and monitoring. They will analyse different of the single market context of EU livestock approaches and then provide guidelines for production, only a comprehensive approach at the the implementation of these strategies in EU level of the EU is likely to be successful. HEALTHY Member States for specific situations at the POULTRY aims at addressing these issues. regional level. The partners will complement their recommendations with a ‘risk assessment toolkit’. A geographic information system (GIS) system will allow policy makers to evaluate the possible consequences of different strategies on the health of poultry flocks, the geographical spread of a disease and economic outcomes of particular interventions.

The results of this project will be disseminated through two important groups: a platform of experts and decision-makers (the ‘users’ of the project results), and representatives of major stakeholders in the poultry business who would be most affected by the implementation of new policies. Close co-operation between these panels and the project research teams could quickly lead to new epizootic disease policies that could circumvent disaster if H5N1 ever gets into poultry 84 stocks. /

Expected results: Potential applications: org . The results of this project will provide a The primary field of application of the results is comprehensive, EU-wide scientific basis for the policy — and decision — making with regard formulation of future EU policies with regard to prevention and control of epizootic poultry to prevention and control of epizootic poultry diseases, i.e. avian influenza, particularly at diseases, particularly AI. the level of the EU and of Member States. Part Coordinator: poultry

Dr Helmut Saatkamp - of the results will also be valuable for other Wageningen University The guidelines for implementation of these stakeholders within the poultry production chains, Dept of Social Sciences Business Economics Chair policies will result in strategies which are more e.g. integrated production chains, animal health (formerly Farm Management tailor made with regard to specific regional services, product boards etc. Group) conditions, e.g. with regard to the density of Hollandseweg 1 6706 Wageningen healthy animals and holdings (i.e. herds), organisation The Netherlands . and trade and structure of livestock production. In Tel: +31 31 74 82 232 [email protected] this way, the veterinary impact of epizootic poultry diseases will be reduced. In turn, the financial and Partners:

Prof. Dr H W Windhorst www economic losses will be largely reduced, as well as and Dr B Grabowski the risks for human health. University of Vechta Institute for Spatial Analysis and Planning of Intensive Implementation of prevention and control Agriculture (ISPA) strategies which comply more with general public Driverstrasse 22 Dr P van Horne 49364 Vechta Agricultural Economics demands on public health, socio-ethical and Germany Research Institute animal welfare issues will lead to an increase Tel: +49 44 41 15 348 LEI-Animal [email protected] P.O. Box 35 in acceptance by the general public of the EU Dr S Marangon and 6700 Wageningen policies in this respect. In turn, it will improve the Dr L Busani The Netherlands livestock sector’s ‘licence to produce’, i.e. its social Istituto Zooprofilattico Tel: +31 31 74 79 761 Sperimentale delle Venezie [email protected] sustainability. Centro Regionale di Dr O Biró Epidemiologia Veterinaria Dept of State Veterinary Viale dell’Università 10 Medicine and Agricultural The delivery of user friendly toolboxes and 35020 Legnaro Economics decision support systems (DSS) will enable Italia Faculty of Veterinary Science Tel: +39 04 98 08 42 55 Szent István University epizootic livestock disease decision makers to [email protected] 1400 Budapest PB 2 evaluate strategy options at the earliest moment Prof. Dr J A Stegeman / Dr Hungary possible, i.e. during the time of development of M Nielen / Dr M Bos Tel: +36 14 78 41 83 Utrecht University [email protected] ideas. Epidemiology Unit Dr V Guberti Dept of Farm Animal Health Istituto Nazionale per la Fauna Faculty of Veterinary Medicine Selvatica P.O. Box 80 163 Via Ca Fornacetta 9 3508 Utrecht 40064 Ozzano E (BO) The Netherlands Italy Tel: +31 30 25 31 013 Tel: +39 05 16 51 22 43

[email protected] [email protected] SSPE-CT-2004-513737 HEALTHY POULTRY HEALTHY Network of Excellence for epizootic disease diagnosis and control

Acronym: EPIZONE EC contribution: €14 000 000 Starting date: 01/06/2006 Key words: Epizootic diseases, network of Duration: 60 months Instrument: Network of Excellence excellence

Summary: strategically driven state-of-art research of world- Aim: Epizootic diseases in agriculture and aquaculture renowned quality. The mission of EPIZONE is to improve research on animals constitute major risks for food production. preparedness, prevention, detection, and control Such diseases spread very fast in high densities of of epizootic diseases within Europe to reduce the susceptible animals through animals, vectors or Problem: economic and social impact of future outbreaks animal products. Outbreaks in Europe have had Epizootic diseases in agriculture and aquaculture of foot-and-mouth disease, classical swine fever, enormous social and economic impact, and need animals constitute major risks for food production. avian influenza and other relevant epizootic to be addressed across the whole production Such diseases spread very fast in high densities diseases like bluetongue and African swine fever, chain of animal-related food. of susceptible animals through animals, vectors through increased excellence by collaboration. or animal products. Outbreaks in Europe showed The objective of EPIZONE is to improve research enormous social and economic impact, and need on preparedness, prevention, detection, and to be addressed across the whole production Expected results: control of epizootics by improvement of excellence chain of animal-related food. EPIZONE will be developed for integration of through collaboration. EPIZONE will be developed scientists in health and production of animals, at for the integration of scientists in health and the European level. Benefits of EPIZONE primarily production of animals, at the European level. concern consumers and stakeholders throughout the food supply chain but also the agriculture The benefits of EPIZONE primarily concern administrations and biotechnology companies. consumers and stakeholders throughout the food supply chain but also the agriculture administrations and biotechnology companies. EPIZONE includes 18 institutes from 12 countries. Partners maintain networks worldwide, linked to EPIZONE, and most have (inter)national reference lab-based tasks for control of epizootics. The management structure of EPIZONE will generate durable interactions between partners. Initially more than 250 key scientists with international reputations, complementary expertise and skills are identified within the partners.

EPIZONE will generate a worldwide network of institutes contributing to available expertise and spreading of excellence. EPIZONE includes the FAO as a world oriented organisation, and an SME specialised in dissemination of knowledge via the Internet. Organisational work packages will develop integration activities, including communications, meetings, and training/ continuous professional development. Scientific work packages will undertake jointly executed research on epizootics selected on importance in Europe and cover four thematic areas: Diagnostics; Intervention Strategies; Surveillance and Epidemiology; Risk Assessment. Given the network structure, the technical resources and 86 the scientific excellence, EPIZONE will assure [email protected] Netherlands The Lelystad AA 8203 2004 Box P.O. Virology of Dept CIDC-Lelystad Control Animal for Disease Institute Central D Coordinator: r Piet van R van r Piet ijn Tel: +44 19 32 35 78 40 78 [email protected] 35 32 19 +44 Tel: UK 3NB KT15 Surrey Addlestone Haw New Agency Laboratories Veterinary D 41 24 [email protected] 23 83 14 +44 Tel: UK 0NF GU24 Surrey Pirbright Road Ash Laboratory Health Pirbright Animal for Institute D 223 [email protected] 17 35 38 +49 Tel: Riems Germany Greifswald-Insel 17493 5a Boddenblick for Health Institute Animal Research Federal Friedrich-Loeffler-Institute B Martin Dr 159 [email protected] 38 02 32 +31 Tel: Netherlands The Lelystad AB 8200 65 Box P.O. BV Lelystad Sciences Group Animal the of Part en Diergezondheid Dierhouderij voor Instituut D Partners: r A D r Linda Poel der van r Wim nthony Fooks nthony ixon eer Tel: +34 91 62 02 300 02 62 91 +34 Tel: Spain Valdeolmos Technology and Research Food and Agriculture for Institute Health National Animal of Center D 705 emmanuel.albina@cirad 93 75 46 +33 Tel: France Montpellier Développement pour le enAgronomique Recherche Centre Internationale de Coopération D 38 43 67 18 +46 Tel: Sweden Uppsala 75189 Anstalt [email protected] [email protected] Agence Française de Sécurité Sécurité de Française Agence D Statens Veterinarmedicinska Veterinarmedicinska Statens D 33 [email protected] 78 34 72 +45 Tel: Denmark Kalvehave 4771 Lindholm Research and Food for Veterinary Institute Danish S Prof afssa.fr 250 p.vannier@ploufragan. 16 60 29 +33 Tel: France Maisons-Alfort 94701 19 Leclerc BP Général Av 27-31 Aliments des Sanitaire r Marisa A r Marisa r E r Philippe Vannier r Philippe r Ulla Carlsson r Ulla mmanuel A mmanuel øren A øren rias lexandersen lbina 06520 Ankara 06520 714) (PK Km 7. Road Eskisehir Ankara Institute FMD D 051 [email protected] 63 88 81 +48 Tel: Poland Partyzantow al Pulawy 24-100 Research Institute Veterinary National Pejsak Zygmunt Prof. 15 25 [email protected] 34 18 93 +86 Tel: China Gansu 730046 Lanzhou 1 Xujiaping CAAS Research Institute Veterinary Lanzhou D 69 43 [email protected] 08 98 04 +39 Tel: Italy (Pd) Legnaro 35020 dell’Università Venezie le V delle Sperimentale Zooprofilattico Istituto D Tel: +32 23 79 05 18 [email protected] 05 79 23 +32 Tel: Belgium Ukkel 1180 99 Groeselenberg Agrochemie en Diergeneeskunde in onderzoek voor Centrum D 277 00 36 [email protected] 87 22 31 +90 Tel: Turkey r Fuat Özyörük r Fuat r Y r I r Frank Koenen r Frank laria Capua laria in Hong in Rome United the Nations of Organization Agriculture and Food D 079 35 [email protected] 59 18 45 +86 Tel: China Harbin nangang st Maduan, 427 CAAS Research Institute Veterinary Harbin D 56 02 [email protected] 29 02 03 +39 Tel: Italy Brescia 25124 9 Bianchi Via Romagna Lombardia dell’Emilia e della Sperimentale Zooprofilattico Istituto D 40 88 [email protected] 53 19 51 +49 Tel: Germany Hannover 30559 17 Bünteweg Virology of Institute Medicine Veterinary of University Moennig Volker Prof. [email protected] 555 65 74 31 +31 Tel: Netherlands The Wageningen 6700 599 Box P.O. Professionals Internet Value Digital O Peter 28 55 [email protected] 05 57 06 +39 Tel: Italy r Keith S r Keith Chen r Hualan r S ylvia Bellini ylvia ude O ude umption phuis

EPIZONE FOOD-CT-2006-016236 www.epizone-eu.net Public health law to support pandemic influenza preparedness

Acronym: PHLawFlu Key words: Law, disease, human rights, public EC contribution: €500 000 Starting date: Planned for 07/2007 health, generic preparedness, pandemic influenza, Duration: 36 months Instrument: Public Health Programme expertise, network

Summary: Problem: Aim: The project consists of two substantive Human pandemic influenza poses global challenges a) To strengthen legal tools in support of disease components. Component 1 will develop a to human health protection. Law is an important preparedness. network of interdisciplinary expertise in public tool in the armoury of states in disease control, b) To support the development of coherent health law. This will be achieved by establishing a but our preliminary research suggests that national European legal responses to the human pandemic Platform for European Public Health Law Expertise laws across Europe are disparate and inadequate influenza threat. by means of a website to facilitate exchange of in addressing disease control and prevention. We c) To develop and support an interdisciplinary data and expertise. The research team will identify have shown that, in their pandemic preparedness network of expertise, knowledge and shared expertise in public health law in Member States, planning, a substantial number of countries experience in public health law in relation to establish a network of expertise and facilitate anticipate the need to reform their public heath communicable disease control, including human communication and exchange by means of laws. Given that the framing of laws by Member pandemic influenza control, across the European conferences, seminars and web contact. States varies considerably, the opportunity exists Union. Component 2 will develop a profile of public for national public health law reforms to result in health law supporting disease preparedness greater coherence. Preparations related to adoption across Europe, by means of case studies focusing of the International Health Regulations also offer on a defined list of public health measures an incentive to reform. The objective of this project related to communicable disease control and is to strengthen Member States’ legal frameworks interventions for the purposes of comparison. in support of pandemic influenza preparedness in a timely manner. While our analysis of Member States’ legislative frameworks will inform the control of generic There is considerable potential for the use of law disease threats, the focus of the case studies as a tool in disease control and prevention. WHO will be on pandemic influenza control. The noted that effective control of the SARS outbreak objective of the cases studies will be to identify in 2003 relied more on traditional control measures commonalities, inconsistencies and gaps in embedded in legislation such as contact tracing and disease control regulation across states. A quarantine, than on modern medical technology. Our database of European laws addressing pandemic preliminary research suggests that there are marked influenza will be developed in English and will be inconsistencies in laws within Europe. Laws may not available through the website. be sufficiently or uniformly grounded in the emerging public health evidence base. Consequently the public health benefits which should accrue through the judicious application of laws may not be fully realised.

Much national public health law addresses nineteenth century perceptions of public health threats and is grounded in nineteenth century mores. Many national disease control laws fail to recognise contemporary issues of human rights and health ethics. We have earlier demonstrated that a substantial number of EU Member States have identified the need to update their domestic legislation in response to the threat of pandemic influenza. This project aims to provide knowledge and build expert capacity which will enable European states to frame laws which assist public health programmes in relation to pandemic influenza, and in so doing to strengthen a coherent Europe-wide 88 approach to pandemic influenza preparedness. disease threats to health. to threats disease non-communicable and communicable all to relation in tool health apublic lawas of use The P Ex c) Dissemination of knowledge and expertise expertise and knowledge of Dissemination c) supporting laws of aprofile of Development b) European maintained Amulti-disciplinary a) applications otential linked to a Platform for European Public Heath Heath Public European for aPlatform to linked law health public on expertise of network legal responses to pandemic threats. pandemic to responses legal coherent develop to seek they as regions global other assist to expertise provide time, over also, may network The lawreform. health public and development policy coherent support and law, health public in challenges health public pragmatic to allied capacity scholarly building in assist EU states, across responses influenza pandemic on knowledge of sharing improved 1, ensure will Component under developed network the through including Dissemination, States. Member across planning preparedness influenza of support lawin health public on laws. national in commonalities and differences identify will and 32 states across quarantine, and measures control border duties, workers’ healthcare closures, school vaccination, example for interventions, and measures health public defined on focus will study The threat. pandemic the to response legal EU-wide coherent and aco-ordinated of formulation the in assist will measures control disease to approaches legal national of Analysis task. this for resource preparedness important an provide will laws control disease of profile The EU account. take lawinto fully plans national that ensuring and EU legislation existing of role and contribution the identifying plans, of evaluations and comparisons making and plans national sharing EU includes the in planning preparedness generic in cooperation for framework The Union. European the across control influenza pandemic human law. health public for relevance has work whose professionals legal and health public of network aEuropean co-ordinate and develop to aims project The Expertise. Law results pected : : [email protected] UK Hertfordshire of Care University Primary Community in and Research for Centre R Prof. Coordinator: obyn Martin obyn [email protected] Lisboa de Portugal Nova Universidade Saúde de Pública Nacional Escola Faria de Lobato Paula Prof. [email protected] Universitaire France Hospitalier Centre A Prof. tu-dresden.de joachim.siegert@mailbox. Dresden Germany Universität Technische D [email protected] UK and Medicine Hygiene Tropical of School London D Partners: r Joachim S r Joachim r R ichard Coker ichard nne Marie D Marie nne iegert uguet

PHLawFlu 800042 European Content for public health awareness of Rural population on Avian Influenza Prevention

Acronym: ECORAIP EC contribution: €199 956 Starting date: 01/04/2007 Key words: Avian flu, awareness campaigns, rural Duration: 18 months Instrument: Public Health Programme population

Summary: Problem: Aim: The population living in the rural areas of Europe In the past decade outbreaks of avian influenza The general objectives of this project are: is crucial to the potential transformation of avian infection among poultry have been reported influenza into a deadly pandemic influenza. This worldwide and have in some cases also led to a) to reduce the risk of human infection in rural project aims to provide practical information and sporadic cases in humans. In Europe a 2003 areas and thus control a pandemic at its onset, guidelines on preventing and managing these outbreak in The Netherlands affected more than by promoting awareness on its risk to the rural potential threats, aiming specifically at the rural 80 people. Avian influenza infection in humans population and proposing preventive measures population, where a gap of information can be can be severe and life-threatening. Highly targeting the rural population’s specific needs. observed. pathogenic avian influenza is a threat to public The measures proposed will be customised to health because it may evolve into an efficient correspond to the particularities of the three The project will take into account the special and dangerous human pathogen. WHO has European areas (central-north, south, eastern); needs of the rural population and its different expressed concern that the avian influenza virus b) to empower European/international/national/ characteristics across three European regions may re-assort its genes with those from a human local initiatives with more efficient tools that which are representative of central-north, influenza virus, thereby acquiring the ability to set the focus on the population living in the south, and eastern Europe. Up-to-date scientific move easily from human to human and thus rural areas of Europe, providing guidelines knowledge will be evaluated and criteria will be triggering a pandemic. for health information campaigns on the defined to integrate or customise the available preparation and prevention of avian influenza material of public health campaigns, so that this pandemics. These guidelines will be developed can be disseminated in a feasible and effective to be specific for the rural population’s way to the population. The channels and characteristics (including animal farming networks used for the dissemination of the public practices) and needs; health material will also be assessed, reviewing c) to increase public health awareness on the existing efforts targeting the rural population. topics of avian flu, and more specifically to Following this review, the most effective fill the existing information gap by promoting communication strategies and information the preparedness of those who are involved in material will be identified and proposed as a best small-scale poultry and general animal farming, practice model. and all those who are more likely to be exposed to wildlife; d) to use this project as a pilot for implementing similar projects which might aim at developing a model for public health campaigns targeting the rural population for health hazards, such as the avian flu pandemic.

90 prevention of avian flu. avian of prevention the for campaigns their designing in this use to expected are who veterinarians, and agronomists experts, disease communicable leaders, municipality authorities, municipality and regional authorities, health public including stakeholders, key to available made be will guideline model The include: will developed be will that guidelines of model The P Ex d) suggestions for different modes of of modes different for suggestions d) material of types different the for suggestions c) avian on acampaign by addressed be to issues b) campaign; the of content the a) and material model the of testing Pilot e) public for material and guidelines Model d) of evaluation and examination Collection, c) public identified the of country per Alist b) related characteristics life rural of Achecklist a) applications otential characteristics which could be related to the the to related be could which characteristics life rural of table the and influenza avian to population. population. target the access successfully to order in acampaign such by used be could These level). municipality the at seminars broadcasting, radio special (i.e. these among choose to how on criteria including dissemination, produced); be not will presentation actual the while developed be will scenarios presentations audiovisual for (i.e. choose to what on criteria and visual) audio, (printed, used be can that prevention; flu analysis. questionnaires’ evaluation the of results the and experience testing pilot the from results the comprising report arelevant of production characteristics. life rural of checklist the against guidelines, relevant the of and campaigns health public of material collected the of evaluation and review the of results the following designed be will material and guidelines model The eastern). south, (central-north, region European each in population rural the for specific prevention, and control epidemic flu avian of topics the on campaigns health this. allow not does budget limited the and project the of scope the isoutside it since done be not will material identified of impact the of analysis aformal Moreover, place. take will translation formal no language, corresponding the comprehend can who experts by evaluated and read be will material identified all Although campaigns. health public of material the flu. avian to regards with population rural the targeting guidelines and campaigns health eastern). south, (central-north, regions European three in epidemic flu avian of spread results pected : : [email protected] Germany Dresden 01061 10 Dresden Helmholtzstr Universität Technische Saxony-Anhalt Public Health Association Research Kirch Wilhelm [email protected] Italy Milan 20157 Grassi B G Via Sacco Universitario L Polo – Ospedaliera Azienda Risk Prevention Health and for Pesticides Centre International A Partners: [email protected] Greece Athens St 11527 Asias Mikras 75 School Medical Athens of University Kapodistrian and National A Coordinator: thena Linos Linos thena ngelo Moretto ngelo for the Environment and Public Public and Health Environment Institute the for International Cyprus D Philip [email protected] Poland Lodz 91-348 St Teresy 8 Occupational Medicine of Institute Nofer Hanke Wojtek [email protected] Cyprus Nicosia 1105 St Iroon Health 5 Public of Harvard School with Association in emokritou

ECORAIP 800123 www.ecoraip.eu Additional classifications

92 Primarily Animal Health Projects:

NOVADUCK, FLUTEST, FLUPATH, NEW-FLUBIRD, FLURESIST, FLU-LAB-NET, FLUTRAIN, AIV VACC DIAGNOSIS, INN-FLU, RIVERS, ConFluTech, HEALTHY POULTRY, LAB-ON-SITE, FLUAID, EPIZONE, AVIFLU, ESNIP, ESNIP 2, ECORAIP Primarily Human Health Projects:

FLUPOL, FLUINNATE, PANFLUVAC, Intranasal H5vaccine, FluVac, EUROFLU, VIRGIL, VIZIER, SARS/FLU VACCINE, UNIVERSAL VACCINE, FLUVACC, RespViruses, CHIMERIC VACCINES, NOVAFLU, FLUPAN, MUCADJ, FLUSECURE, EISS, PHLawFlu Research Framework Programme 5 (FP5): MUCADJ, ESNIP, FLUPAN, AVIFLU, NOVAFLU Research Framework Programme 6 (FP6): FLUPOL, FLUINNATE, PANFLUVAC, Intransasal H5vaccine, FluVac, EUROFLU, NOVADUCK, FLUTEST, FLUPATH, NEW-FLUBIRD, FLURESIST, FLU-LAB-NET, FLUTRAIN, AIV VACC DIAGNOSIS, INN-FLU, RIVERS, ConFluTech, VIRGIL, HEALTHY POULTRY, LAB-ON-SITE, VIZIER, SARS/FLU VACCINE, UNIVERSAL VACCINE, FLUVACC, FLUAID, EPIZONE, RespViruses, CHIMERIC VACCINES, ESNIP 2 Programme of community action in the field of Public Health: FLUSECURE, EISS, PHLawFlu, ECORAIP Projects focusing (almost) exclusively on influenza: FLUPOL, FLUINNATE, PANFLUVAC, Intranasal H5vaccine, FluVac, EUROFLU, NOVADUCK, FLUTEST, FLUPATH, NEW-FLUBIRD, FLURESIST, FLU-LAB-NET, FLUTRAIN, AIV VACC DIAGNOSIS, INN-FLU, RIVERS, ConFluTech, UNIVERSAL VACCINE, FLUVACC, FLUAID, CHIMERIC VACCINES, NOVAFLU, AVIFLU, FLUPAN, ESNIP, ESNIP 2, MUCADJ, FLUSECURE, EISS, ECORAIP Projects addressing a broader range of (viral or other infectious) diseases, but with a significant part devoted to influenza: VIRGIL (large network dealing with antiviral drug resistance in influenza, hepatitis B and hepatitis C), HEALTHY POULTRY (dedicated to control of all epizootic poultry diseases), LAB-ON-SITE (diagnostic tests for nine animal diseases, including avian influenza), VIZIER (structural characterisation of a diverse set of viruses, including influenza), RespViruses (immune response of the elderly to respiratory viruses) and EPIZONE (large network dealing with epizootic diseases), SARS/FLU VACCINE (combined vaccine against SARS and influenza), PHLawFlu (general public health law but with a focus on pandemic influenza) INDEX

94 6///////////////////////////////////////////////////////////////////////////////////////////////////////////////////Vaccines 36///////////////////////////////////////////////////////////////////////////////// Diagnostics and Surveillance 56////////////////////////////////////////////////////////////////////////////////////////////////////////////////// Biology 72/////////////////////////////////////////Networking, Training, Socio-Economic and Legal Issues 16...... Aiv Vacc Diagnosis 48...... Aviflu 26...... Chimeric Vaccines 78...... ConFluTech 90...... Ecoraip 54...... EISS 86...... Epizone 50...... Esnip 52...... Esnip 2 62...... Euroflu 74...... Flu-Lab-Net 22...... Fluaid 60...... Fluinnate 30...... Flupan 64...... Flupath 58...... Flupol 42...... Fluresist 34...... Flusecure 38...... Flutest 76...... Flutrain 12...... Fluvac 24...... Fluvacc 84...... Healthy Poultry 66...... Inn-Flu 10...... IntranasalH5vaccine 46...... Lab-on-Site 32...... Mucadj 40...... New-Flubird 14...... Novaduck 28...... Novaflu 8...... Panfluvac 88...... PHLawFlu 70...... RespViruses 44...... Rivers 18...... Sars Flu Vaccine 20...... Universal Vaccine 80...... Virgil 68...... Vizier

European Commission

EUR 22822 — Influenza Research — EU funded projects 2001-2007

Luxembourg: Office for Official Publications of the European Communities

2007 — 96 pp. — 21.0 x 29.7 cm

ISBN 978-92-79-05420-4

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interdisciplinary cooperation in the area of zoonoses and emerging infectious diseases. Commission’s Framework Programmes for Research, but also through its Public Health Programmes. It demonstrates It Programmes. Health Public its through also but Research, for Programmes Framework Commission’s moreresponserapiditsthe as to well field,as thisresearch fundingin EU ofrecord trackterm longscope and the recent developments. The inclusion of both animal and human health projects underlines the importance of human infections with virusanHPAI H5N1 makes pandemic influenza a paradigm of the potential health threat that emerging infectious diseases pose the to world. The present catalogue assembles 38 projects funded between 2001 and 2007 mostly through the European pandemic outbreaks, such as the 'Spanish Flu' which took more than 50 million lives in 1918-1919. Birds are natural are Birds 1918-1919. in lives million 50 than more took which 'Spanish Flu' the outbreaks,pandemicas such reservoirs of influenza viruses and highly pathogenic variants of these avian influenza viruses (HPAI) economiclosses.poultrycanincreasingspreadingof highnumberepizoonoticspopulations, Therapidly in to cause leading Everyseasonalyear influenza thoughtisresponsiblebe to dangerousmorefor500than 000more deaths worldwide.much addition,In to lead to potential the have virusescirculating the of antigenssurface the in changes major