DOCSLIB.ORG

Infection Both in Vitro and in Vivo Mycobacterium Tuberculosis Control

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Infection Both in Vitro and in Vivo Mycobacterium Tuberculosis Control CD4+ T Cells Mediate IFN-γ-Independent Control of Mycobacterium tuberculosis Infection Both In Vitro and In Vivo This information is current as Siobhán C. Cowley and Karen L. Elkins of September 28, 2021. J Immunol 2003; 171:4689-4699; ; doi: 10.4049/jimmunol.171.9.4689 http://www.jimmunol.org/content/171/9/4689 Downloaded from References This article cites 43 articles, 30 of which you can access for free at: http://www.jimmunol.org/content/171/9/4689.full#ref-list-1 Why The JI? Submit online. http://www.jimmunol.org/ • Rapid Reviews! 30 days* from submission to initial decision • No Triage! Every submission reviewed by practicing scientists • Fast Publication! 4 weeks from acceptance to publication *average by guest on September 28, 2021 Subscription Information about subscribing to The Journal of Immunology is online at: http://jimmunol.org/subscription Permissions Submit copyright permission requests at: http://www.aai.org/About/Publications/JI/copyright.html Email Alerts Receive free email-alerts when new articles cite this article. Sign up at: http://jimmunol.org/alerts The Journal of Immunology is published twice each month by The American Association of Immunologists, Inc., 1451 Rockville Pike, Suite 650, Rockville, MD 20852 Copyright © 2003 by The American Association of Immunologists All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. The Journal of Immunology CD4؉ T Cells Mediate IFN-␥-Independent Control of Mycobacterium tuberculosis Infection Both In Vitro and In Vivo Siobha´n C. Cowley1 and Karen L. Elkins1 Although IFN-␥ is necessary for survival of Mycobacterium tuberculosis infection in people and animal models, it may not be sufficient to clear the infection, and IFN-␥ is not a reliable correlate of protection. To determine whether IFN-␥-independent mechanisms of immunity exist, we developed a murine ex vivo culture system that directly evaluates the ability of splenic or lung lymphocytes to control the growth of M. tuberculosis within infected macrophages, and that models in vivo immunity to tuber- ␥-culosis. Surprisingly, CD4؉ T cells controlled >90% of intracellular M. tuberculosis growth in the complete absence of IFN stimulation of macrophages, via a NO-dependent mechanism. Furthermore, bacillus Calmette-Guerin-vaccinated IFN-␥-deficient Downloaded from mice exhibited significant protection against M. tuberculosis challenge that was lost upon depletion of CD4؉ T cells. These findings demonstrate that CD4؉ T cells possess IFN-␥-independent mechanisms that can limit the growth of an intracellular pathogen and are dominant in secondary responses to M. tuberculosis. The Journal of Immunology, 2003, 171: 4689–4699. uberculosis is a significant global health problem, as one- macrophages (11). These findings, along with the extreme suscep- third of the world’s population is estimated to be infected tibility of people (12) and mice (13, 14) with disruptions in the http://www.jimmunol.org/ T with Mycobacterium tuberculosis, and 8 million new ac- IFN-␥ or p40 gene to M. tuberculosis infection, has led to pro- tive cases occur annually. The only current vaccine, live Myco- posed use of IFN-␥ as a correlate of protection for new vaccines bacterium bovis bacillus Calmette-Guerin (BCG),2 is of variable against tuberculosis (15). efficacy in different geographic regions, ranging from as little as However, numerous examples in the literature indicate that the 0% to as much as 80% protection (1). However, development of levels of IFN-␥ produced by a mouse in response to a candidate improved vaccine candidates is limited by our incomplete under- vaccine do not always correlate with the effectiveness of that vac- standing of the cellular and molecular immune mechanisms re- cine during M. tuberculosis challenge (16). Similarly, a recent in- quired for the elimination of a M. tuberculosis infection (2). vestigation evaluated the efficacy of human BCG vaccination using M. tuberculosis is a facultative intracellular pathogen that sur- several assays and found that mycobacterial growth inhibition did by guest on September 28, 2021 vives and grows primarily within macrophages in the host. In both not correlate with IFN-␥ responses (17). Furthermore, mycobac- mice and humans, the immune response has difficulty eliminating terial infection of human and mouse macrophages disrupts a piv- a M. tuberculosis infection, resulting in the establishment of a otal part of IFN-␥ intracellular signaling, resulting in inhibition of latent infection in humans and a chronic disease state in mice. IFN-␥-induced gene expression (18, 19). It also remains unclear Similar to other intracellular pathogens, immunity to M. tubercu- whether the activity of IFN-␥ in vivo is primarily bactericidal or losis is cell mediated. Murine studies using in vivo Ab depletion, ␥ gene-disrupted mice, or adoptive transfer indicate that CD4ϩ T only bacteriostatic. Thus, although IFN- is essential for the de- cells are key to the control of infection (3–7). Mice with disrup- velopment of an immune response that prolongs the life span of an ␤ infected animal, it is not sufficient to eliminate an M. tuberculosis tions in the genes for 2-microglobulin, TAP, or CD8 are more susceptible to M. tuberculosis than wild-type (WT) mice, also im- infection, as mice infected with M. tuberculosis develop a long- plicating a role for CD8ϩ T cells and the MHC class I pathway term chronic infection. (8–10). Both CD4ϩ and CD8ϩ T cells have been shown to pro- We are interested in investigating the other important immune duce IFN-␥ during an M. tuberculosis infection (2). In experimen- mechanisms that contribute to the control of M. tuberculosis in- tal models, IL-12 has a central role in regulating Th1 T cell pro- tracellular growth, particularly because understanding these mech- duction of IFN-␥, and IFN-␥ is clearly key in the activation of anisms may provide improved correlates of protection for vaccine research. To more readily discover IFN-␥-independent mecha- nisms, we have developed a murine ex vivo culture system that Laboratory of Mycobacterial Diseases and Cellular Immunology, Division of Bacte- rial, Parasitic, and Allergenic Products, Center for Biologics Evaluation and Research/ directly evaluates the ability of splenic or lung lymphocytes to Food and Drug Administration, Rockville, MD 20852 control the growth of M. tuberculosis within infected macro- Received for publication May 6, 2003. Accepted for publication August 21, 2003. phages, and that reflects the in vivo components of the immune The costs of publication of this article were defrayed in part by the payment of page system known to be involved in controlling an M. tuberculosis charges. This article must therefore be hereby marked advertisement in accordance infection. Using this system, we demonstrate that a very substan- with 18 U.S.C. Section 1734 solely to indicate this fact. tial proportion of the ability of CD4ϩ T cells to control M. tuber- 1 Address correspondence and reprint requests to Dr. Siobha´n Cowley or Dr. Karen Elkins, LMDCI/DBPAP/CBER/FDA, 1401 Rockville Pike, HFM 431, Rockville, MD culosis growth occurs in the absence of IFN-␥ stimulation of the 20852. E-mail address: [email protected] or [email protected] macrophage. Furthermore, we show that IFN-␥-deficient mice vac- 2 Abbreviations used in this paper: BCG, bacillus Calmette-Guerin; BMM␾, bone cinated with BCG exhibit limited but readily demonstrable pro- ␥ marrow-derived macrophage; cDMEM, complete DMEM; GKO, KO; KO, knock- tection against a subsequent virulent M. tuberculosis aerosol chal- out; MOI, multiplicity of infection; PPD, purified protein derivative; TB, tuberculosis; ϩ ϩ WT, wild type. lenge that is dependent on CD4 T cells. Thus, CD4 T cells have Copyright © 2003 by The American Association of Immunologists, Inc. 0022-1767/03/$02.00 4690 IFN-INDEPENDENT CONTROL OF M. tuberculosis BY CD4ϩ T CELLS IFN-␥-independent mechanisms that can affect control of M. tu- libitum) for 2–3 mo to eliminate the BCG infection (22). This reduced the berculosis growth during secondary challenge. BCG numbers in the spleens and lungs to undetectable levels, as deter- mined 1 wk postremoval of the antibiotic treatment; however, at later time points (ϳ30 days), regrowth of BCG was evident in the spleens of the Materials and Methods GKO animals. To generate a population of Listeria-specific T cells, mice Animals were sublethally infected with 104 L. monocytogenes strain EGD intrad- ermally, and spleens were harvested 4–6 wk later. Six- to twelve-week-old male specific pathogen-free C57BL6/J, 129S1/ ␥ SvImJ, or IFN- R-deficient mice on a C57BL6/J background were pur- Depletion of T cell subsets in vivo chased from The Jackson Laboratory (Bar Harbor, ME). Animals were housed in sterile microisolator cages in a barrier environment at the Center In vivo depletion of CD4ϩ cells in BCG-vaccinated mice was performed for Biologics Evaluation and Research. Mice were fed autoclaved food and by i.p. administration of 500 ␮g of the anti-CD4 Ab GK1.5 twice weekly water ad libitum. All experiments were performed under protocols ap- for the duration of the experiment. Successful in vivo depletion of CD4 proved by the Center for Biologics Evaluation and Research/Food and cells was confirmed (Ͻ0.2% positive cells remaining) in the spleens of Drug Administration Institutional Animal Care and Use Committee. depleted animals using flow cytometry (see below). Culture and infection of bone marrow-derived macrophage Harvesting and enrichment of splenocytes (BMM␾) with bacteria
Load more

Recommended publications
  • Meta Analyses – Pros and Cons
    Meta analyses – pros and cons Emily S Sena, PhD Centre for Clinical Brain Sciences, University of Edinburgh @camarades_ CAMARADES: Bringing evidence to translational medicine CAMARADES • Collaborative Approach to Meta-Analysis and Review of Animal Data from Experimental Studies • Look systematically across the modelling of a range of conditions • Data Repository – 30 Diseases – 40 Projects – 25,000 studies – from over 400,000 animals CAMARADES: Bringing evidence to translational medicine Why do we do meta-analysis of animal studies? • Animal models are generally performed to inform human health but when should you be convinced to move to the next step? • Systematic reviews & meta-analyses: – assess the quality and range of evidence – identify gaps in the field – quantify relative utility of outcome measures – inform power/sample size calculations – assess for publication bias – try to explain discrepancies between preclinical and clinical trial results – inform clinical trial design CAMARADES: Bringing evidence to translational medicine Systematic review and meta-analysis • Pros – What have we learnt about…….. • Translation? • Quality? • The 3Rs? • Cons – What are the limitations? • As good as the data that goes in? • Rapidly outdated • The Impact CAMARADES: Bringing evidence to translational medicine Systematic review and meta-analysis • Pros – What have we learnt about…….. • Translation? • Quality? • The 3Rs? • Cons – What are the limitations? • As good as the data that goes in? • Rapidly outdated • The Impact CAMARADES: Bringing evidence
    [Show full text]
  • Melanoma: Epidemiology, Risk Factors, Pathogenesis, Diagnosis and Classification
    in vivo 28: 1005-1012 (2014) Review Melanoma: Epidemiology, Risk Factors, Pathogenesis, Diagnosis and Classification MARCO RASTRELLI1, SAVERIA TROPEA1, CARLO RICCARDO ROSSI2 and MAURO ALAIBAC3 1Melanoma and Sarcoma Unit, Veneto Institute of Oncology, IOV- IRCCS, Padova, Italy; 2Melanoma and Sarcoma Unit, Veneto Institute of Oncology, IOV-IRCCS and Department of Surgery, Oncology and Gastroenterology, University of Padova, Padova, Italy; 3Dermatology Unit, University of Padova, Padova, Italy Abstract. This article reviews epidemiology, risk factors, Epidemiology pathogenesis and diagnosis of melanoma. Data on melanoma from the majority of countries show a rapid At the start of 21st century, melanoma remains a potentially increase of the incidence of this cancer, with a slowing of fatal malignancy. At a time when the incidence of many the rate of incidence in the period 1990-2000. Males are tumor types is decreasing, melanoma incidence continues to approximately 1.5-times more likely to develop melanoma increase (1). Although most patients have localized disease than females, while according to other studies, the different at the time of the diagnosis and are treated by surgical prevalence in both sexes must be analyzed in relation with excision of the primary tumor, many patients develop age: the incidence rate of melanoma is grater in women metastases (2). than men until they reach the age of 40 years, however, by The incidence of malignant melanoma has been increasing 75 years of age, the incidence is almost 3-times as high in worldwide, resulting in an important socio-economic men versus women. The most important and potentially problem. From being a rare cancer one century ago, the modifiable environmental risk factor for developing average lifetime risk for melanoma has now reached 1 in 50 malignant melanoma is the exposure to ultraviolet (UV) in many Western populations (3).
    [Show full text]
  • Adaptive Enrichment Designs in Clinical Trials
    Annual Review of Statistics and Its Application Adaptive Enrichment Designs in Clinical Trials Peter F. Thall Department of Biostatistics, M.D. Anderson Cancer Center, University of Texas, Houston, Texas 77030, USA; email: [email protected] Annu. Rev. Stat. Appl. 2021. 8:393–411 Keywords The Annual Review of Statistics and Its Application is adaptive signature design, Bayesian design, biomarker, clinical trial, group online at statistics.annualreviews.org sequential design, precision medicine, subset selection, targeted therapy, https://doi.org/10.1146/annurev-statistics-040720- variable selection 032818 Copyright © 2021 by Annual Reviews. Abstract All rights reserved Adaptive enrichment designs for clinical trials may include rules that use in- terim data to identify treatment-sensitive patient subgroups, select or com- pare treatments, or change entry criteria. A common setting is a trial to Annu. Rev. Stat. Appl. 2021.8:393-411. Downloaded from www.annualreviews.org compare a new biologically targeted agent to standard therapy. An enrich- ment design’s structure depends on its goals, how it accounts for patient heterogeneity and treatment effects, and practical constraints. This article Access provided by University of Texas - M.D. Anderson Cancer Center on 03/10/21. For personal use only. first covers basic concepts, including treatment-biomarker interaction, pre- cision medicine, selection bias, and sequentially adaptive decision making, and briefly describes some different types of enrichment. Numerical illus- trations are provided for qualitatively different cases involving treatment- biomarker interactions. Reviews are given of adaptive signature designs; a Bayesian design that uses a random partition to identify treatment-sensitive biomarker subgroups and assign treatments; and designs that enrich superior treatment sample sizes overall or within subgroups, make subgroup-specific decisions, or include outcome-adaptive randomization.
    [Show full text]
  • Use of in Vivo and in Vitro Models to Guide Dose Selection for Neonatal Infections
    Use of In Vivo and In Vitro Models to Guide Dose Selection for Neonatal Infections Professor William Hope Antimicrobial Pharmacodynamics & Therapeutics University of Liverpool September 2016 Disclosures • William Hope has received research funding from Pfizer, Gilead, Astellas, AiCuris, Amplyx, Spero Therapeutics and F2G, and acted as a consultant and/or given talks for Pfizer, Basilea, Astellas, F2G, Nordic Pharma, Medicines Company, Amplyx, Mayne Pharma, Spero Therapeutics, Auspherix, Cardeas and Pulmocide. First of all BIOMARKER OUTCOME OF CLINICAL DOSE INTEREST/IMPORTANCE •Decline in Log10CFU/mL •Linked to an outcome of clinical interest •Survival 7 6 5 4 CFU/g) CNS 10 3 Effect (log 2 1 0 0.01 0.1 1 10 100 1000 Total doseDose 5FC (mg/kg) PHARMACOKINETICS PHARMACODYNAMICS Concept of Expensive Failure COST Derisking White Powder Preclinical Program Clinical Program Concept from Trevor Mundell & Gates Foundation And this… Normal Therapeutics Drug A, Dose A1 versus Drug B, Dose B1 Pharmacodynamics is the Bedrock of ALL Therapeutics…it sits here without being seen One of the reasons simple scaling does not work is the fact pharmacodynamics are different Which means the conditions that govern exposure response relationships in neonates need to be carefully considered …Ignore these at your peril Summary of Preclinical Pharmacodynamic Studies for Neonates** • Hope el al JID 2008 – Micafungin for neonates – Primary question of HCME – Rabbit model with hematogenous dissemination • Warn et al AAC 2010 – Anidulafungin for neonates – Primary question
    [Show full text]
  • Role and Limitations of Epidemiology in Establishing a Causal Association Eduardo L
    Seminars in Cancer Biology 14 (2004) 413–426 Role and limitations of epidemiology in establishing a causal association Eduardo L. Franco a,∗, Pelayo Correa b, Regina M. Santella c, Xifeng Wu d, Steven N. Goodman e, Gloria M. Petersen f a Departments of Epidemiology and Oncology, McGill University, 546 Pine Avenue West, Montreal, QC, Canada H2W1S6 b Department of Pathology, Louisiana State University Health Sciences Center, New Orleans, LA, USA c Department of Environmental Health Sciences, Mailman School of Public Health, Columbia University, New York, NY, USA d Department of Epidemiology, University of Texas MD Anderson Cancer Center, Houston, TX, USA e Department of Biostatistics, Bloomberg School of Public Health, Johns Hopkins University, Baltimore, MD, USA f Department of Health Sciences Research, Mayo Clinic College of Medicine, Rochester, MN, USA Abstract Cancer risk assessment is one of the most visible and controversial endeavors of epidemiology. Epidemiologic approaches are among the most influential of all disciplines that inform policy decisions to reduce cancer risk. The adoption of epidemiologic reasoning to define causal criteria beyond the realm of mechanistic concepts of cause-effect relationships in disease etiology has placed greater reliance on controlled observations of cancer risk as a function of putative exposures in populations. The advent of molecular epidemiology further expanded the field to allow more accurate exposure assessment, improved understanding of intermediate endpoints, and enhanced risk prediction by incorporating the knowledge on genetic susceptibility. We examine herein the role and limitations of epidemiology as a discipline concerned with the identification of carcinogens in the physical, chemical, and biological environment. We reviewed two examples of the application of epidemiologic approaches to aid in the discovery of the causative factors of two very important malignant diseases worldwide, stomach and cervical cancers.
    [Show full text]
  • (GIVIMP) for the Development and Implementation of in Vitro Methods for 829 Regulatory Use in Human Safety Assessment
    1 2 Draft GUIDANCE DOCUMENT ON GOOD IN VITRO METHOD PRACTICES (GIVIMP) 3 FOR THE DEVELOPMENT AND IMPLEMENTATION OF IN VITRO METHODS FOR 4 REGULATORY USE IN HUMAN SAFETY ASSESSMENT 5 6 FOREWORD 7 8 A guidance document on Good In Vitro Method Practices (GIVIMP) for the development and 9 implementation of in vitro methods for regulatory use in human safety assessment was 10 identified as a high priority requirement. The aim is to reduce the uncertainties in cell and 11 tissue-based in vitro method derived predictions by applying all necessary good scientific, 12 technical and quality practices from in vitro method development to in vitro method 13 implementation for regulatory use. 14 The draft guidance is coordinated by the European validation body EURL ECVAM and has 15 been accepted on the work plan of the OECD test guideline programme since April 2015 as a 16 joint activity between the Working Group on Good Laboratory Practice (GLP) and the 17 Working Group of the National Coordinators of the Test Guidelines Programme (WNT). 18 The draft document prepared by the principal co-authors has been sent in September 2016 to 19 all 37 members of the European Union Network of Laboratories for the Validation of 20 Alternative Methods (EU-NETVAL1) and has been subsequently discussed at the EU- 21 NETVAL meeting on the 10th of October 2016. 22 By November/December 2016 the comments of the OECD Working Group on GLP and 23 nominated experts of the OECD WNT will be forwarded to EURL ECVAM who will 24 incorporate these and prepare an updated version.
    [Show full text]
  • Transforming Clinical Trials with Digital Tech, and What That Means for the Patient
    ■ SCRIP 100 SPONSORED BY: Transforming Clinical Trials With Digital Tech, And What That Means For The Patient Many stakeholders within the pharma and biotech industries are management systems, safety systems and data repositories. A witnessing radical shifts taking place in how clinical trials are central data storage location provides sponsors and sites access conceived, designed and conducted. This transformation relies in real time, and increases productivity by allowing information heavily on applying the power of digital technologies. to be quickly shared and managed in a secure fashion. As new technologies emerge, they will converge through networks Additionally, the continuous streaming of data to cloud-based plat- and cloud-based platforms to create a new digital health care ecosys- forms could accelerate clinical trials and decrease protocol amend- tem. Collectively, they will have a greater impact on clinical trials than ments, resulting in reduced clinical trial costs. Also, sponsors can use any one technology would achieve separately. At the center of this cloud-based platforms for data submission to regulatory agencies, ecosystem is the patient, demonstrated by the uptick in personalized which has the potential to accelerate drug development, streamline therapies and a growing emphasis on patient-reported outcomes. regulatory review and enhance regulatory decision-making.2 While the mean projected return on new drug research and devel- DEMOCRATIZATION OF AI, DATA AND ALGORITHMS opment (R&D) investments by a dozen large cap biopharma firms fell from 10.1% in 2010 to 1.9% in 2018,1 an opportunity remains The influx of big data is fueling algorithms that are the building blocks for emerging digital technologies to improve R&D productivity.
    [Show full text]
  • Psychological Approaches in the Treatment of Specific Phobias: a Meta-Analysis ☆ ⁎ Kate B
    Available online at www.sciencedirect.com Clinical Psychology Review 28 (2008) 1021–1037 Psychological approaches in the treatment of specific phobias: A meta-analysis ☆ ⁎ Kate B. Wolitzky-Taylor, Jonathan D. Horowitz, Mark B. Powers 1, Michael J. Telch Laboratory for the Study of Anxiety Disorders, Department of Psychology, The University of Texas at Austin, United States Received 21 August 2007; received in revised form 11 February 2008; accepted 27 February 2008 Abstract Data from 33 randomized treatment studies were subjected to a meta-analysis to address questions surrounding the efficacy of psychological approaches in the treatment of specific phobia. As expected, exposure-based treatment produced large effects sizes relative to no treatment. They also outperformed placebo conditions and alternative active psychotherapeutic approaches. Treatments involving in vivo contact with the phobic target also outperformed alternative modes of exposure (e.g., imaginal exposure, virtual reality, etc.) at post-treatment but not at follow-up. Placebo treatments were significantly more effective than no treatment suggesting that specific phobia sufferers are moderately responsive to placebo interventions. Multi-session treatments marginally outperformed single-session treatments on domain-specific questionnaire measures of phobic dysfunction, and moderator analyses revealed that more sessions predicted more favorable outcomes. Contrary to expectation, effect sizes for the major comparisons of interest were not moderated by type of specific phobia. These findings provide the first quantitative summary evidence supporting the superiority of exposure-based treatments over alternative treatment approaches for those presenting with specific phobia. Recommendations for future research are also discussed. © 2008 Elsevier Ltd. All rights reserved. Keywords: Specific phobia; Meta-analysis; Exposure treatment Contents 1.
    [Show full text]
  • AS04 Data Sciences Project (Educating for the Future Working Group)
    Paper AS04 Data Sciences Project (Educating for the Future Working Group) Sascha Ahrweiler, PHUSE, Wuppertal, Germany Aldir Medeiros Filho, London, United Kingdom ABSTRACT According to Google Trends, the word “Data Science” is currently at a peak interest for worldwide searches. Companies like Uber and Amazon have built entire business models using data science methodology. The healthcare sector has also seen new players especially in consumer devices, which increased awareness of healthy lifestyles by applying advanced analytics. The pharmaceutical industry is adapting to these new technologies. FDA approved or cleared devices are already used mainly for exploratory purposes in clinical trials and create huge volumes of data. We can create valuable insights when we connect these new data sources to clinical data and apply Data Science methods like Machine Learning, Deep Learning or Artificial Intelligence. This paper discusses the Data Sciences Project within Educating for the Future Working Group, with a view to educate the broader PHUSE community in data science techniques in healthcare so they can be prepared in dealing with new challenges. INTRODUCTION This paper introduces the scope of work of Data Sciences Project within the PHUSE Educating for the Future (EftF) Working Group, why we decided to first focus our attention on the clinical drug program development domain as well and tentatively delineate and differentiate the potential role of a Clinical Data Scientist within this domain. The PHUSE Working Group, “Educating for the Future“(EftF) was initiated prior to the PHUSE CS Symposium (CSS) event in 2018. The goal of this EftF Working Group is to keep up with the evolving industry and to educate the PHUSE community at large on relevant topics.
    [Show full text]
  • Making Sense of Animal Research
    Making sense of animal research REPLACE REDUCE REFINE Drug Development and the 3Rs Number of Development Phase Questions Addressed Duration Tools and 3Rs Principles Applied Compounds Tested Discovery of New Drug What target enzyme or protein? What test system to show drug-protein 3 years 1000‘000 Non animal research: interaction? What are chemical lead structures? Computer modeling High throughput screening Imaging techniques Optimization What is structure-activity relationship? What molecules have not only 2-3 years 10‘000 In vitro studies: intrinsic activity but also drug-like properties (absorbability, metabolic Cell cultures stability)? TissuesOrgans 100 Preclinical Development How can kg-amounts efficiently be synthesized? What are suitable 1-2 years 5 Animal and in vitro tests: formulations? What is pharmacokinetic metabolic and safety profile Ethical review; Improved housing and care; of the drug candidate? Refined sampling techniques; Smarter study design; Imaging technologies; Chip technology; Microdosing; Stem cells; International harmonization; Replacing higher by lower species; etc… Clinical Testing What is tolerable dose range? What is dose-effect relationship? 4-5 years 1-2 Human clinical trials: Is drug sufficiently safe at efficacious doses? Phase I, II and III (safety, efficacy, variability) Registration What are sources of variability? What is adverse effect profile? 1 year 0,1 The pace of medical research is fast, but there are still many health challenges ahead. Animals continue to play an invaluable role in meeting these challenges - both in experimental research, and in ensuring maximum safety of treatments before their use in hu- mans. The process for developing a medicine does not start with animal intro research, which represents only about 10% of the whole R&D proc- ess.
    [Show full text]
  • Reproducibility and Rigor in Animal-Based Research
    ILAR Journal, 2020, Vol. 60, No. 1, 17–23 doi: 10.1093/ilar/ilz015 Advance Access Publication Date: 5 November 2019 Review Article Reproducibility and Rigor in Animal-Based Research Downloaded from https://academic.oup.com/ilarjournal/article-abstract/60/1/17/5612731 by guest on 15 May 2020 Malcolm Macleod1,* and Swapna Mohan2 1Centre for Clinical Brain Sciences, University of Edinburgh, Edinburgh, United Kingdom; and 2National Institutes of Health, 8600 Rockville Pike, Bethesda, Maryland (previously in the Division of Policy and Education at the NIH Office of Laboratory Animal Welfare, NIH). *Corresponding Author: Malcolm R .Macleod, PhD FRCP, Centre for Clinical Brain Sciences, University of Edinburgh, 49 Little France Crescent, Edinburgh EH16 4TJ, United Kingdom. E-mail: [email protected]. Abstract Increasing focus on issues of research reproducibility affords us the opportunity to review some of the key issues related in vivo research. First, we set out some key definitions, to guide the reader through the rest of the paper. Next we consider issues of epistemology, of how animal experiments lead to changes in our understanding of biomedicine and, potentially, to the development of new therapeutics. Here we consider the meaning of statistical significance; the importance of understanding whether findings have general truth; and the advances in knowledge which can result from ‘failed’ replication. Then, we consider weaknesses in the design, conduct and reporting of experiments, and review evidence for this from systematic reviews and from experimental studies addressing these issues. We consider the impact that these weaknesses have on the development of new treatments for human disease, and reflect on the response to these issues from the biomedical research community.
    [Show full text]
  • Role of Epidemiology in the Prevention of Adverse Human
    William Halperin, MD MPH DrPH Jun Tashiro, BA (MD/MPH expected May 2011) National Nanotechnology Initiative: Human and Environmental Exposure Assessment Workshop February 24- 25, 2009 Charge • Plenary –Characterize health of exposed populations and environments (biota) • Breakout discussions –Identify emerging needs –Build a dialogue toward consensus –Identify ongoing projects Outline 1. Cascade of prevention 2. Role of Surveillance 3. Meta-analysis of literature 4. Meta-analysis of research in progress 5. Meta-analysis of grant opportunities 6. Identification of holes 7. Future directions Basics of Prevention: The Cascade of Prevention Design Toxicologic Testing Substitution/Elimination Engineering Controls Environ. Exposure Monitoring Personal Exposure Monitoring Biological Monitoring Medical Screening Clinical Care Rehabilitation Accommodation References What is Surveillance and Where is it in the Cascade? • “Ongoing systemic collection, analysis, and dissemination of exposure and health data on groups of workers for the purpose of early detection of disease and injury as well as patterns of occurrence presumably leading to prevention of subsequent disease.” Source: Schulte P, Geraci C, Zumwalde R, et al. Occupational risk management of engineered nanoparticles. Journal of Occupational & Environmental Hygiene. Apr 2008;5(4):239-249. Where is Surveillance? Design Toxicologic Testing Substitution/Elimination Engineering Controls Environ. Exposure Monitoring Personal Exposure Monitoring Biological Monitoring Medical Screening Clinical
    [Show full text]