Novel Protein and Poxvirus-Based Vaccine Combinations for Simultaneous Induction of Humoral and Cell-Mediated Immunity This information is current as Claire L. Hutchings, Sarah C. Gilbert, Adrian V. S. Hill and of September 24, 2021. Anne C. Moore J Immunol 2005; 175:599-606; ; doi: 10.4049/jimmunol.175.1.599 http://www.jimmunol.org/content/175/1/599 Downloaded from References This article cites 50 articles, 25 of which you can access for free at: http://www.jimmunol.org/content/175/1/599.full#ref-list-1 http://www.jimmunol.org/ Why The JI? Submit online. • 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 by guest on September 24, 2021 *average 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 © 2005 by The American Association of Immunologists All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. The Journal of Immunology Novel Protein and Poxvirus-Based Vaccine Combinations for Simultaneous Induction of Humoral and Cell-Mediated Immunity1 Claire L. Hutchings, Sarah C. Gilbert, Adrian V. S. Hill, and Anne C. Moore2 The presence of both cell-mediated and humoral immunity is important in protection from and clearance of a number of infectious pathogens. We describe novel vaccine regimens using combinations of plasmid DNA, poxvirus and protein to induce strong Ag-specific T cell and Ab responses simultaneously in a murine model. Intramuscular (i.m.) immunization with plasmid DNA encoding the middle Ag of hepatitis B (DNA) concurrently with a commercial hepatitis B virus (HBV) vaccine (Engerix-B) followed by boosting immunizations with both modified vaccinia virus Ankara (MVA) encoding the middle Ag of HBV and Engerix-B induced high levels of CD4؉ and CD8؉ T cells and high titer Ab responses to hepatitis B surface Ag (HbsAg). Substitution of Downloaded from Engerix-B with adjuvant-free rHBsAg induced similar T cell responses and greatly enhanced Ab levels. Repeated immunizations with recombinant or nonrecombinant MVA mixed with Ag induced higher titers of Abs compared with immunization with either Ag or Engerix-B further demonstrating this novel adjuvant effect of MVA. The poxviruses NYVAC, fowlpox (FP9) and ALVAC, and to a lesser extent, adenovirus, also displayed similar adjuvant properties when used in combination with rHBsAg. The use of poxviruses as an adjuvant for protein to concurrently induce Ag-specific T cells and Abs could be applied to the development of vaccines for many diseases, including HIV and malaria, where both cell mediated and humoral immunity may be important for http://www.jimmunol.org/ protection. The Journal of Immunology, 2005, 175: 599–606. he major aim of vaccination is to generate sufficiently sponse is detectable in individuals with acute self-limited HBV but strong immune responses of the correct nature to protect is absent in individuals with chronic infection (10–12). This sug- T against disease. Most vaccines that are in used in humans, gests that a cell-mediated immune response may be important in except bacillus Calmette-Gue´rin, have been developed by optimiz- eliminating virus at an early stage of infection. A vaccine that ing their capacity to induce Ab responses. Novel candidate vac- effectively induces both cellular and humoral immunity may im- cines that have been developed to date either induce immune re- prove the efficacy of a HBV vaccination. sponses that are dominated by cellular or humoral immune Recombinant DNA and viral vectors have been used for a num- by guest on September 24, 2021 responses. For example, vaccines against intracellular pathogens, ber of years in laboratory animals and in human clinical trials as such as HIV or liver stage malaria, have been designed either to vectors for the Ag(s) of interest to induce strong effector and mem- induce strong T cell responses (1–3) or to induce Ab responses ory CD4ϩ and CD8ϩ T cell responses in both the prevention and against the extracellular form of the virus or parasite (4, 5). How- treatment of disease (2, 13–16). Widely used viral vectors include ever, it is now widely believed that the generation of both cellular modified vaccinia virus Ankara (MVA) and NYVAC, both atten- and humoral responses may be important in vaccine efficacy for uated forms of vaccinia virus, the avipoxviruses FP9 and ALVAC, diverse causes of disease including viruses, bacteria, parasites, and and also nonreplicating adenovirus (ADV) (2, 15–21). We and cancer (4, 6–8). The licensed prophylactic vaccine to hepatitis B others have previously demonstrated in a pre-erythrocytic malaria virus (HBV),3 Engerix-B, is an example of a vaccine that induces vaccine model, where induction of IFN-␥ secreting T cells corre- Ab responses. It induces protective Abs in Ͼ85% of healthy re- cipients (9) and comprises the small surface Ag, S (HBsAg), pro- lates with protection, that heterologous prime-boost immunization duced in yeast cells (Saccharomyces cerevisiae) adsorbed to the with DNA and viral vectors is protective in mice, primate and adjuvant aluminum phosphate (alum). A vigorous CD8ϩ CTL re- human challenge models (2, 13, 16, 22, 23, 50). With respect to HBV vaccines, DNA and MVA encoding the middle Ag of HBV, comprising S and Pre-S2 Ags, have been designed in this labora- Wellcome Trust Centre for Human Genetics, University of Oxford, Oxford, United tory as a potential therapeutic vaccine for chronic HBV infection. Kingdom This prime-boost regimen has demonstrated potent induction of Received for publication January 30, 2004. Accepted for publication April 29, 2005. specific T cells in mice and humans and is currently in phase II The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked advertisement in accordance clinical trials (J. Schneider, personal communication). with 18 U.S.C. Section 1734 solely to indicate this fact. In this study, we examined methods of inducing strong cellular 1 This work was supported by the Wellcome Trust. A.V.S.H. is a Wellcome Trust and Ab responses in a mouse model of candidate and licensed Principal Fellow. HBV vaccines. Although this model cannot be used to assess pro- 2 Address correspondence and reprint requests to Dr. Anne Moore, Wellcome Trust tection against HBV challenge, immunogenicity of Engerix-B can Centre for Human Genetics, University of Oxford, Roosevelt Drive, Oxford, OX3 7BN, U.K. E-mail address: [email protected]. be tested in mice, and this clinically licensed vaccine functions as 3 Abbreviations used in this paper: HBV, hepatitis B virus; HBsAg, small hepatitis B a good standard to compare our candidate vaccines to. We report surface Ag; MVA, modified vaccinia virus Ankara; ADV, adenovirus; ADVnr, non- here that prime-boost immunization with DNA and MVA (D/M) recombinant adenovirus; CMI, cell-mediated immunity; DC, dendritic cell.; LN, lymph node; DLN, draining LN; FP, fowlpox; i.d., intradermal; SFC, spot-forming induces potent T cell responses but poor levels of Ab against the cells. encoded HBsAg. Conversely, repeat immunization with Engerix-B Copyright © 2005 by The American Association of Immunologists, Inc. 0022-1767/05/$02.00 600 ENHANCED IMMUNOGENICITY BY COADMINISTRATION OF POXVIRUS AND PROTEIN induced a weak T cell response but greatly increased Abs to rH- Aldrich) to each well. Optical densities at 405 nm were measured for each BsAg compared with D/M. The concurrent administration of these well. End point titers were determined at the x-axis intercept of the dilution curve, at three times the absorbance (A ) given for naive mouse serum vaccines induced both T cells and Abs to HBsAg. We sought to 405 diluted from 1/200 accordingly. analyze and further improve this coinduction of cell-mediated im- munity (CMI) and humoral immunity by combining the viral vec- Statistical analysis tors MVA, FP, ALVAC NYVAC, and ADV with the main com- Statistical analysis was performed using SPSS for Windows version 10. ponent of Engerix-B, rHBsAg. Here we show that poxviruses can Mann-Whitney U tests were used to determine differences between groups. enhance T cell and Ab responses to a coadministered protein while retaining strong T cell immunogenicity for their encoded recom- Results binant Ag thereby demonstrating a method of strongly inducing Immunogenicity of DNA/MVA.HBs and protein immunization both types of immunity against a target Ag. This combination vac- T cell and Ab responses induced by the prime-boost regime of cine approach may be of benefit to the development of efficacious DNA/MVA were compared with homologous protein immuniza- vaccines where this type of immunity is required to protect against tion with or without alum (Fig. 1). pathogen challenge. Immunization with DNA/MVA induced high levels of IFN-␥ producing cells against peptide encoding a dominant CD8ϩ Materials and Methods epitope and whole rHBsAg in the spleen (Fig. 1A) and facial DLN Animals and immunizations (Fig. 1B). Immunization with HBsAg or Engerix-B induced min- Female BALB/c (H-2d) mice (BMSU, John Radcliffe Hospital, Oxford, imal peptide and rHBsAg responses in spleen. However, the pep- U.K.), 4–6 wk old, were used in all experiments. Before immunization tide-specific response induced in axial DLN when immunized s.c. Downloaded from mice were anesthetized i.p.