SMALLPOX VACCINE A M EDICAL D ICTIONARY , B IBLIOGRAPHY , AND A NNOTATED R ESEARCH G UIDE TO I NTERNET R E FERENCES
J AMES N. P ARKER , M.D. AND P HILIP M. P ARKER , P H .D., E DITORS
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ICON Health Publications ICON Group International, Inc. 4370 La Jolla Village Drive, 4th Floor San Diego, CA 92122 USA Copyright 2004 by ICON Group International, Inc. Copyright 2004 by ICON Group International, Inc. All rights reserved. This book is protected by copyright. No part of it may be reproduced, stored in a retrieval system, or transmitted in any form or by any means, electronic, mechanical, photocopying, recording, or otherwise, without written permission from the publisher. Printed in the United States of America. Last digit indicates print number: 10 9 8 7 6 4 5 3 2 1
Publisher, Health Care: Philip Parker, Ph.D. Editor(s): James Parker, M.D., Philip Parker, Ph.D. Publisher's note: The ideas, procedures, and suggestions contained in this book are not intended for the diagnosis or treatment of a health problem. As new medical or scientific information becomes available from academic and clinical research, recommended treatments and drug therapies may undergo changes. The authors, editors, and publisher have attempted to make the information in this book up to date and accurate in accord with accepted standards at the time of publication. The authors, editors, and publisher are not responsible for errors or omissions or for consequences from application of the book, and make no warranty, expressed or implied, in regard to the contents of this book. Any practice described in this book should be applied by the reader in accordance with professional standards of care used in regard to the unique circumstances that may apply in each situation. The reader is advised to always check product information (package inserts) for changes and new information regarding dosage and contraindications before prescribing any drug or pharmacological product. Caution is especially urged when using new or infrequently ordered drugs, herbal remedies, vitamins and supplements, alternative therapies, complementary therapies and medicines, and integrative medical treatments. Cataloging-in-Publication Data Parker, James N., 1961Parker, Philip M., 1960Smallpox Vaccine: A Medical Dictionary, Bibliography, and Annotated Research Guide to Internet References / James N. Parker and Philip M. Parker, editors p. cm. Includes bibliographical references, glossary, and index. ISBN: 0-597-84630-8 1. Smallpox Vaccine-Popular works. I. Title.
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Disclaimer This publication is not intended to be used for the diagnosis or treatment of a health problem. It is sold with the understanding that the publisher, editors, and authors are not engaging in the rendering of medical, psychological, financial, legal, or other professional services. References to any entity, product, service, or source of information that may be contained in this publication should not be considered an endorsement, either direct or implied, by the publisher, editors, or authors. ICON Group International, Inc., the editors, and the authors are not responsible for the content of any Web pages or publications referenced in this publication.
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Acknowledgements The collective knowledge generated from academic and applied research summarized in various references has been critical in the creation of this book which is best viewed as a comprehensive compilation and collection of information prepared by various official agencies which produce publications on smallpox vaccine. Books in this series draw from various agencies and institutions associated with the United States Department of Health and Human Services, and in particular, the Office of the Secretary of Health and Human Services (OS), the Administration for Children and Families (ACF), the Administration on Aging (AOA), the Agency for Healthcare Research and Quality (AHRQ), the Agency for Toxic Substances and Disease Registry (ATSDR), the Centers for Disease Control and Prevention (CDC), the Food and Drug Administration (FDA), the Healthcare Financing Administration (HCFA), the Health Resources and Services Administration (HRSA), the Indian Health Service (IHS), the institutions of the National Institutes of Health (NIH), the Program Support Center (PSC), and the Substance Abuse and Mental Health Services Administration (SAMHSA). In addition to these sources, information gathered from the National Library of Medicine, the United States Patent Office, the European Union, and their related organizations has been invaluable in the creation of this book. Some of the work represented was financially supported by the Research and Development Committee at INSEAD. This support is gratefully acknowledged. Finally, special thanks are owed to Tiffany Freeman for her excellent editorial support.
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About the Editors James N. Parker, M.D. Dr. James N. Parker received his Bachelor of Science degree in Psychobiology from the University of California, Riverside and his M.D. from the University of California, San Diego. In addition to authoring numerous research publications, he has lectured at various academic institutions. Dr. Parker is the medical editor for health books by ICON Health Publications. Philip M. Parker, Ph.D. Philip M. Parker is the Eli Lilly Chair Professor of Innovation, Business and Society at INSEAD (Fontainebleau, France and Singapore). Dr. Parker has also been Professor at the University of California, San Diego and has taught courses at Harvard University, the Hong Kong University of Science and Technology, the Massachusetts Institute of Technology, Stanford University, and UCLA. Dr. Parker is the associate editor for ICON Health Publications.
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About ICON Health Publications To discover more about ICON Health Publications, simply check with your preferred online booksellers, including Barnes&Noble.com and Amazon.com which currently carry all of our titles. Or, feel free to contact us directly for bulk purchases or institutional discounts: ICON Group International, Inc. 4370 La Jolla Village Drive, Fourth Floor San Diego, CA 92122 USA Fax: 858-546-4341 Web site: www.icongrouponline.com/health
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Table of Contents FORWARD .......................................................................................................................................... 1 CHAPTER 1. STUDIES ON SMALLPOX VACCINE ................................................................................ 3 Overview........................................................................................................................................ 3 Federally Funded Research on Smallpox Vaccine.......................................................................... 3 E-Journals: PubMed Central ....................................................................................................... 28 The National Library of Medicine: PubMed ................................................................................ 28 CHAPTER 2. CLINICAL TRIALS AND SMALLPOX VACCINE ............................................................. 43 Overview...................................................................................................................................... 43 Recent Trials on Smallpox Vaccine.............................................................................................. 43 Keeping Current on Clinical Trials ............................................................................................. 47 CHAPTER 3. BOOKS ON SMALLPOX VACCINE................................................................................. 49 Overview...................................................................................................................................... 49 Book Summaries: Online Booksellers........................................................................................... 49 Chapters on Smallpox Vaccine..................................................................................................... 49 CHAPTER 4. PERIODICALS AND NEWS ON SMALLPOX VACCINE ................................................... 51 Overview...................................................................................................................................... 51 News Services and Press Releases................................................................................................ 51 Academic Periodicals covering Smallpox Vaccine ....................................................................... 55 APPENDIX A. PHYSICIAN RESOURCES ............................................................................................ 59 Overview...................................................................................................................................... 59 NIH Guidelines............................................................................................................................ 59 NIH Databases............................................................................................................................. 61 Other Commercial Databases....................................................................................................... 63 APPENDIX B. PATIENT RESOURCES ................................................................................................. 65 Overview...................................................................................................................................... 65 Patient Guideline Sources............................................................................................................ 65 Finding Associations.................................................................................................................... 68 APPENDIX C. FINDING MEDICAL LIBRARIES .................................................................................. 71 Overview...................................................................................................................................... 71 Preparation................................................................................................................................... 71 Finding a Local Medical Library.................................................................................................. 71 Medical Libraries in the U.S. and Canada ................................................................................... 71 ONLINE GLOSSARIES.................................................................................................................. 77 Online Dictionary Directories ..................................................................................................... 77 SMALLPOX VACCINE DICTIONARY ...................................................................................... 79 INDEX .............................................................................................................................................. 105
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FORWARD In March 2001, the National Institutes of Health issued the following warning: "The number of Web sites offering health-related resources grows every day. Many sites provide valuable information, while others may have information that is unreliable or misleading."1 Furthermore, because of the rapid increase in Internet-based information, many hours can be wasted searching, selecting, and printing. Since only the smallest fraction of information dealing with smallpox vaccine is indexed in search engines, such as www.google.com or others, a non-systematic approach to Internet research can be not only time consuming, but also incomplete. This book was created for medical professionals, students, and members of the general public who want to know as much as possible about smallpox vaccine, using the most advanced research tools available and spending the least amount of time doing so. In addition to offering a structured and comprehensive bibliography, the pages that follow will tell you where and how to find reliable information covering virtually all topics related to smallpox vaccine, from the essentials to the most advanced areas of research. Public, academic, government, and peer-reviewed research studies are emphasized. Various abstracts are reproduced to give you some of the latest official information available to date on smallpox vaccine. Abundant guidance is given on how to obtain free-of-charge primary research results via the Internet. While this book focuses on the field of medicine, when some sources provide access to non-medical information relating to smallpox vaccine, these are noted in the text. E-book and electronic versions of this book are fully interactive with each of the Internet sites mentioned (clicking on a hyperlink automatically opens your browser to the site indicated). If you are using the hard copy version of this book, you can access a cited Web site by typing the provided Web address directly into your Internet browser. You may find it useful to refer to synonyms or related terms when accessing these Internet databases. NOTE: At the time of publication, the Web addresses were functional. However, some links may fail due to URL address changes, which is a common occurrence on the Internet. For readers unfamiliar with the Internet, detailed instructions are offered on how to access electronic resources. For readers unfamiliar with medical terminology, a comprehensive glossary is provided. For readers without access to Internet resources, a directory of medical libraries, that have or can locate references cited here, is given. We hope these resources will prove useful to the widest possible audience seeking information on smallpox vaccine. The Editors
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From the NIH, National Cancer Institute (NCI): http://www.cancer.gov/cancerinfo/ten-things-to-know.
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CHAPTER 1. STUDIES ON SMALLPOX VACCINE Overview In this chapter, we will show you how to locate peer-reviewed references and studies on smallpox vaccine.
Federally Funded Research on Smallpox Vaccine The U.S. Government supports a variety of research studies relating to smallpox vaccine. These studies are tracked by the Office of Extramural Research at the National Institutes of Health.2 CRISP (Computerized Retrieval of Information on Scientific Projects) is a searchable database of federally funded biomedical research projects conducted at universities, hospitals, and other institutions. Search the CRISP Web site at http://crisp.cit.nih.gov/crisp/crisp_query.generate_screen. You will have the option to perform targeted searches by various criteria, including geography, date, and topics related to smallpox vaccine. For most of the studies, the agencies reporting into CRISP provide summaries or abstracts. As opposed to clinical trial research using patients, many federally funded studies use animals or simulated models to explore smallpox vaccine. The following is typical of the type of information found when searching the CRISP database for smallpox vaccine: •
Project Title: A SAFER AND MORE EFFICACIOUS SMALLPOX VACCINE Principal Investigator & Institution: Yilma, Tilahun D.; Professor and Director; Interntl Lab Molecular Biology; University of California Davis Sponsored Programs, 118 Everson Hall Davis, Ca 95616 Timing: Fiscal Year 2003; Project Start 30-SEP-2003; Project End 31-JAN-2006
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Healthcare projects are funded by the National Institutes of Health (NIH), Substance Abuse and Mental Health Services (SAMHSA), Health Resources and Services Administration (HRSA), Food and Drug Administration (FDA), Centers for Disease Control and Prevention (CDCP), Agency for Healthcare Research and Quality (AHRQ), and Office of Assistant Secretary of Health (OASH).
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Summary: (provided by applicant): In the aftermath of the attacks of September 11 and the anthrax scare, we have a heightened awareness of US vulnerability to bioterrorism. One of the most feared infectious agents is variola virus, the causative agent of smallpox. Various strains of vaccinia virus (VV) are highly effective in preventing this disease, but have definite rates of complications. Severe illness or death is rare in people with normal immune responses, but considerably more common in individuals with cell-mediated immune defects. The number of individuals that are at risk from this normally innocuous vaccine has greatly increased with the spread of the human immunodeficiency virus (HIV), and it now becomes important to improve the efficacy and safety of this vaccine. We have worked extensively with VV as a recombinant vaccine for a number of diseases; our rinderpest vaccine was described as one of two outstanding rVVs in a leading journal (G. Ada, Nature 349:369, 1991). We have also developed strategies for attenuating VV while enhancing efficacy, with one of the most effective being the incorporation of the interferon-gamma (IFN-gamma) gene. We have shown that expression of IFN-gamma leads to a TH1 immune response essential against viral infection with no deleterious effects. We have also studied the effects of inactivating VV immunomodulating genes such as B8R, B13R, and B22R that are virulence factors in VV. Based on our past experience, we propose developing a safer and more efficacious vaccine for smallpox based on the New York City Board of Health (Wyeth) strain of VV that is currently used in the US. We will delete the B8R gene and insertionally inactivate the TK virulence gene with the human IFN-gamma gene to increase attenuation of the virus and the protective cell-mediated immune responses. This recombinant VV will be compared to the parental vaccine in both normal and simian immunodeficiency virus-infected macaques (used as a model for HIV-infected humans) to assess efficacy and safety for normal and immunodeficient individuals. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •
Project Title: ASSESSMENT OF IMMUNITY TO VACCINIA AND MVA Principal Investigator & Institution: Dolin, Raphael C.; Maxwell Finland Professor of Medicine; Dana-Farber Cancer Institute 44 Binney St Boston, Ma 02115 Timing: Fiscal Year 2003; Project Start 01-SEP-2003; Project End 31-AUG-2008 Summary: Despite its extraordinary effectiveness against smallpox infection, the parameters of protective immunity generated by vaccinia remain incompletely understood. Large numbers of individuals are at high risk for serious toxicities if vaccinated with or exposed to vaccinia, and the need to develop safer vaccines against smallpox underscores the importance of understanding the immunologic basis for such protection. This project will undertake a detailed examination of T and B cell responses to immunization with vaccinia and MVA under carefully controlled conditions in normal subjects, and to MVA in patients with atopic dermatitis or patients with hematologic malignancies and immune deficiency after hematopoietic stem cell transplantation. The studies will measure neutralizing and other binding antibodies, lymphoproliferative responses by 3H Thymidine incorporation, cytotoxic T-cell activity by a 51Cr release assay, and T-cell gamma interferon responses by ELISPOT. Normal subjects who were vaccinated with either vaccinia or MVA, will be challenged with vaccinia to determine whether protection against challenge was conferred. Measurements of immune responses will be conducted sequentially at multiple time points, including those before and after challenge. Rates and patterns of responses wilt be compared between vaccinia and MVA recipients, and between normals and the patient cohorts. The specificity of the B and T cell responses will be examined employing bioinformatics approaches and molecular techniques of the Research Resource Technical
Studies
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Development Component of the Center. These studies will measure antibody responses to a panel of recombinant vaccinia-encoded proteins identified by bioinformatics as potentially important in neutralization, including the EGF-like domain of DIL (SPGF). Since all subjects in our initial studies will be HLA-A*0201 positive, we will be able to utilize peptides from the recombinant vaccinia proteins to characterize class I allele specific T-cell responses by ELISPOT, cytokine flow cytometry, and peptide-MHC tetramers. Non-class I allele specific T-cell responses will be examined using whole recombinant proteins. Spectratype analysis will also be used to identify predominant Tcell epitopes. These studies wilt provide a comprehensive picture of the immune responses to vaccination with vaccinia and MVA, and identify epitopes against which important responses are directed. These studies will also demonstrate whether 2 groups of high-risk patients respond to MVA in a similar fashion to normals. The proposed studies in years 1 and 2 focus on MVA, but similar approaches would be utilized to study other candidate vaccines against smallpox, as well as vaccines against other agents important in biodefense. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •
Project Title: ASSESSMENT OF SMALLPOX SPECIFIC IMMUNE RESPONSES Principal Investigator & Institution: Denny, Thomas N.; Assistant Professor of Pathology,; Pediatrics; Univ of Med/Dent Nj Newark Newark, Nj 07103 Timing: Fiscal Year 2003; Project Start 30-SEP-2002; Project End 31-AUG-2004 Summary: This proposal, in response to RFA-AI-02-002, seeks funding to study longlasting immunity (e.g. immunological memory or recall) to smallpox (variola) in individuals who were previously vaccinated in the United States commercially approved smallpox vaccine (preparation of live Vaccinia virus). In the aftermath of September 11, 2001, it is now critical to better define mechanisms of smallpox protective immunity or disease resistance in the general population and in those who may be classified immune compromised and therefore, considered at a higher risk of infection. Though smallpox was considered a disease of historical interest only, since its earlier eradication, it as been known to be a significant pathogen for potential bioterrorist activities. Investigation of long-lasting immunity using current state-of-the-art techniques or methodologies may help determine which individuals are better prepared to serve as "first-line" responders. In addition, this information may help determine a better strategy for use of limited, non-universally administrable vaccine material available today. Specific Aim 1: To assess smallpox-specific long-lasting immunologic responses and correlate to donor age and time since last vaccination in healthy individuals that have previously received a smallpox vaccination. Hypothesis: Initial childhood vaccine administration will be associated with differing levels of smallpoxspecific host immune responses that will vary with number of immunizations given and the time since last vaccination. Specific Aim 2: To assess smallpox-specific long-lasting immunologic responses in HIV- and HIV+ individuals through collaboration with the NIAID DAIDS ACTG and correlate to age and infection status in individuals previously having smallpox vaccination. Hypothesis: HIV infection, CD4 or viral load status, time since last vaccination and host age will each be associated differing levels of smallpoxspecific host immune responses. Specific Aim 3: To viral epitope map and evaluate the CD4/CD8 immune subset critical memory component of virus-specific cell-mediated responses in healthy and immune comprised study participants. Herd immunity or natural boosting is absent from this model, it represents a unique opportunity to extensively study pathogen specific immunological memory. Hypothesis: Response
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level and/or frequency of CD4 or CD8 cells to various immunodominant viral epitopes will change with age, time since last vaccination or presence of disease (e.g, HIV). Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •
Project Title: AUGMENTING INNATE AND VACCINE IMMUNE RESPONSE WITH DER-G Principal Investigator & Institution: Zimmerman, Daniel H.; Cel-Sci Corporation 8229 Boone Blvd, Ste 802 Vienna, Va 22182 Timing: Fiscal Year 2003; Project Start 30-SEP-2003; Project End 31-MAR-2005 Summary: (provided by applicant): Concerns regarding the United States' vulnerability to a terrorist attack with smallpox have stimulated development of new vaccines, prophylactics and therapeutics. However, a new and successful smallpox vaccine intervention program, or one to treat exposed individual requires not only an effective vaccine, but also agents to treat the complications arising from a widespread vaccination program. Recently, we found that derG, an N terminal deamidated analogue of the human MHC II beta-chain (aa135-149) showed significant protective activity in infectious disease models of malaria and HSV and adjuvant activity for vaccines. The primary goals of this application are to determine the protective activity of derG for vaccinia infections, identify its mechanism of action to support its' development as an immunotherapeutic and/or prophylactic for vaccinia and smallpox infections as a single agent and as an adjuvant for vaccinia vaccines. The ultimate goal of these Phase I studies is to develop sufficient data to justify studies in a primate model of efficacy and animal safety and toxicology studies to support human clinical trials. In addition to the concern that smallpox may be used as a weapon of mass destruction, recombinant vaccinia viruses (VV) are also used as vaccines or as vectors for immunotherapy resulting in the recommendation that individuals working with VV vectors be vaccinated. However, the present vaccines, when available, are associated with serious complications in approximately 1:10,000 primary vaccinees, and mortality in about 1:10 6 primary vaccinations. Our strategy is to use derG as prophylaxsis and as an adjuvant for a replication incompetent vaccine prime to limit the toxicity by live vaccinia virus and to target mucosal activity. Thus, these studies will deliver derG and the vaccine by intranasal administration and will monitor both systemic and mucosal (pulmonary) immunity. We propose to use derG, together with an infectious, but replication incompetent VV (irradiated) vaccine to examine the hypothesis that immune augmentation with derG will prolong survival and also provide adjuvant activity for a replication incompetent vaccine, thereby providing an additional reduction in viral burden and prolongation of survival for vaccinia-infected mice as a model of smallpox infection. This hypothesis will be tested with the following Specific Aims: 1: Determine the immunoregulatory and adjuvant properties of derG during VV vaccination of mice. 2: Determine the immune augmenting and adjuvant activity of derG in mice sub-lethally infected with vaccinia as a model of smallpox. 3: Determine the therapeutic potential of immune intervention in the treatment of lethal vaccinia virus infections and identify surrogates of therapeutic activity. In these studies we will follow survival and ovarian viral titers after intranasal challenge with VV. The studies will also analyze the mechanism of action to identify immune surrogates for use in clinical studies. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen
Studies
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Project Title: BACTERIAL DEVELOPMENT
ANTIGENS
AND
ANTHRAX
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Principal Investigator & Institution: Hewlette, Erik; University of Maryland Balt Prof School Baltimore, Md 21201 Timing: Fiscal Year 2003; Project Start 04-SEP-2003; Project End 28-FEB-2008 Summary: The Middle Atlantic Region consortium proposes to establish a Regional Center of Excellence for Biodefense and Emerging Infectious Diseases Research (RCE), whose theme is "Defense against Bioterror and Emerging Infection Agents." The proposed Research Projects are: 1) Anthrax (new Bacillus anthracis antigens, tested in animal models; compounds to impede anthrax infection; mouse model for imaging germination and bacterial distribution; development of a mucosal live vector prime/parenteral boost anthrax vaccine); 2) Hemorrhagic Fever and other Emerging Viruses (identification/characterization of neutralizing human monoclonal antibodies reactive to functionally important determinants on Henipaviruses, Bunyaviruses, West Nile, Ebola and Marburg); 3) Poxviruses (subunit variola vaccine; identification of new targets of neutralizing antibody and of vaccinia immune globulin; and development of a mouse ectromelia virus model of smallpox pathogenesis/prevention); 4) Tularemia (conjugate tularemia vaccine; study possible therapy of F. tularensis infection using reagents already under clinical testing for sepsis; evaluate currently available bisphosphonate drugs as a first line of defense for individuals exposed to F. tularensis; attenuated, live-vector tularemia vaccine); 5) Low-Dose Enteric Pathogens (role of type 1 Cryptosporidium parvum candidate genes in pathogenesis/susceptibility to infection, as a prelude to vaccine development; Shigella dysenteriae 1 and EHEC vaccines; novel therapeutics for EHEC disease; diagnostics for detection of these pathogens in water, food, and environmental specimens); 6) Public Health Response Research (needle-free immunization and vaccine-adjuvanting strategies; immunogenetics of human immune response to smallpox vaccine; innovative diagnostic platforms for routine clinical use and in known or suspected bioterror events). Three Career Development Projects (to train the next generation of biodefense investigators) and 4 Developmental Projects (high-risk projects on biodefense agents) will be funded per year. Training will include a Media Training Course; a short course in "GMP Production and Process Development" (in collaboration with Aventis Pasteur Vaccines and Merck Vaccines); a Category A Bioterror Agent Clinical Surveillance Course; a "hands-on" course on working in BSL-3 facilities; and travel awards for RCE scientists and trainees to visit other RCE labs to learn techniques or perform collaborative experiments. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •
Project Title: CELL-MEDIATED IMMUNE RESPONSES TO VACCINIA VIRUSES Principal Investigator & Institution: Crowe, James E.; Associate Professor of Pediatrics; Pediatrics; Vanderbilt University 3319 West End Ave. Nashville, Tn 372036917 Timing: Fiscal Year 2003; Project Start 15-SEP-2003; Project End 28-FEB-2007 Summary: (provided by applicant): Detailed analysis of T cell responses from individuals enrolled in vaccinia virus vaccine trials may dramatically improve our understanding of the effects of dilution on vaccine immunogenicity and on the relationship of cell-mediated immunity to protection from adverse events following immunization. Our experiments will begin to address the level of cell-mediated immunity elicited in naive adults following vaccination with the Aventis Pasteur smallpox vaccine. Furthermore, these studies will establish a paradigm in which researchers can begin to incorporate improved measures of cell-mediated immunity in
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clinical environments, generating data that will provide useful surrogate biomarkers of immune responses related to adverse events or protection. Specifically, we will test the hypothesis that the lack of a vigorous response of host T cells to immunodominant cytolytic T cell epitopes following primary immunization is associated with adverse events that are related to failure to clear virus shedding rapidly. Previous work examining cell-mediated immunity to vaccinia virus during clinical vaccine trials is very limited. Small studies have examined the effects of smallpox vaccination and identified that human CTL memory responses to vaccinia virus do occur. Much of the work performed to date examining CM1 responses in humans to specific antigens/viruses have been performed using bulk culture proliferation techniques and standard cytotoxicity assays. While these assay provide a measure of T cell responsiveness to specific antigens, they fail to delineate which of the subsets of T cells are involved in protective memory responses or are associated with adverse events. We will examine many of these questions by taking advantage of new technology that allows for examination of T cell responses at the single cell level, and in an immunodominant epitope specific manner. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •
Project Title: CELLULAR IMMUNITY TO CATEGORY A-C VIRUSES IN HUMANS Principal Investigator & Institution: Ennis, Francis A.; Professor; None; Univ of Massachusetts Med Sch Worcester Office of Research Funding Worcester, Ma 01655 Timing: Fiscal Year 2003; Project Start 30-SEP-2003; Project End 31-MAR-2008 Summary: (provided by applicant): The proposed UMASS Center for Translational Research on Human Immunology and Biodefense is a comprehensive, interdepartmental collaboration to address, as its overall scientific theme, the role of human T lymphocyte responses in the immunopathogenesis of and protection from category A-C viral pathogens. The Center encompasses senior and junior investigators with significant prior expertise in human immunology and research on biodefense pathogens, including translation to clinical studies, and is organized around the following components: Project 1 (Poxviruses) will define immunodominant human T cell epitopes of vaccinia virus as markers of vaccine efficacy and advance knowledge toward the development of improved smallpox vaccines. Project 2 (Hantaviruses) will define human immune responses associated with immunopathogenesis of hemorrhagic fever with renal syndrome. Project 3 (Flaviviruses) will define protective and immunopathological cross-reactive human T cell responses to sequential virus infections. The Technical Development Component (TDC) will develop novel solidphase array and proteomics technologies for application in T cell epitope identification, detection and enumeration of virus-specific T cells, and identification of biomarkers of protective or pathological T cell responses. Core facilities will provide flow cytometry, MHC class I production, MHC class II production, and program administration services for use by the research projects and TDC. A Pilot Projects component will support pilot funding of promising novel research concepts proposed by UMMS faculty. An Education component will support short-term training of non-Center investigators. The proposed Center will address important NIAID research priorities related to these biodefense pathogens as well as career development for young investigators. The Center will facilitate synergistic interactions between the various investigators, to be assessed by periodic internal and external review that will greatly enhance the overall research productivity. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen
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Project Title: CUTANEOUS IMMUNITY AND VACCINIA Principal Investigator & Institution: Kupper, Thomas S.; Professor and Chair; DanaFarber Cancer Institute 44 Binney St Boston, Ma 02115 Timing: Fiscal Year 2003; Project Start 01-SEP-2003; Project End 31-AUG-2008 Summary: The overarching goal of this project is to understand the mechanisms by which transepidermal inoculation of human skin with vaccinia (scarification) leads to a protective immune response to smallpox, and to use this knowledge to help develop vaccination strategies that are both safe and effective, particularly for patients who currently are not vaccination candidates (e.g., patients with atopic dermatitis). Studies will be performed using both human and murine model systems. The potential for vaccinia to productively infect both normal and atopic skin tissue, cells, and artificial skin constructs, will be assessed by vanous techniques. In collaboration with investigators from Project 1, both skin and blood will be sampled at various time points from vaccinated normal volunteers. We will test the validity of our paradigm of cutaneous immune response, wherein Langerhans cells containing virus fragments from infected epidermis migrate to draining lymph nodes and differentiate into potent mature dendritic cells and activate naive T cells. These T cells expand clonally and differentiate into central memory and skin-homing effector memory T cells. Effector memory T cells extravasate from dermal vessels at the vaccine site and enter the papillary dermis and epidermis. Central memory cells traffic into secondary lymphoid tissues and provide long-term immunologic memory. We will characterize key cellular and humoral elements of the protective immune response to variola generated as a result of these events. We will test the extent to which atopic dermatitis patients and normal volunteers vaccinated with MVA develop similar key elements of this protective response. In murine models, we will manipulate the cutaneous microenvironment with biological response modifiers and determine whether these maneuvers improve the immune response to vaccinia. Transgenic mice with targeted epidermal expression of cytokines and chemokines that influence dendritic cell migration and function will also be studied. The efficacy of vaccination will be assessed by testing resistance to vaccinia challenge in these mice after prior vaccination with MVA or vaccinia, with a major goal being to enhance vaccination efficiency. The Harvard Skin Disease Research Center has studied both innate and acquired immune response mechanisms in skin for more than 15 years. The resources of the HSDRC will provide a rich environment for these studies and will enhance collaborative interactions with investigators leading projects 1, 2, and 4. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen
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Project Title: DEFENSE AGAINST BIOWARFARE AND EMERGING INFECTION AGENTS Principal Investigator & Institution: Levine, Myron Max.; Director; Medicine; University of Maryland Balt Prof School Baltimore, Md 21201 Timing: Fiscal Year 2003; Project Start 04-SEP-2003; Project End 28-FEB-2008 Summary: (provided by applicant): The Middle Atlantic Region consortium proposes to establish a Regional Center of Excellence for Biodefense and Emerging Infectious Diseases Research (RCE) whose theme is "Defense Against Bioterror and Emerging Infection Agents." The proposed Research Projects are: 1) Anthrax (new Bacillus anthracis antigens tested in animal models; compounds to impede anthrax infection; mouse model for imaging germination and bacterial distribution; and development of a mucosal live vector prime/parenteral boost anthrax vaccine); 2) Hemorrhagic Fever and
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Smallpox Vaccine
other Emerging Viruses (identification/characterization of neutralizing human monoclonal antibodies reactive to functionally important determinants on Henipaviruses, Bunyaviruses, West Nile, Ebola and Marburg viruses); 3) Poxviruses (subunit variola vaccine; identification of new targets of neutralizing antibody and of vaccinia immune globulin; and development of a mouse ectromelia virus model of smallpox pathogenesis/prevention); 4) Tularemia (conjugate tularemia vaccine; study possible therapy of Francisella tularensis infection using reagents already under clinical testing for sepsis; evaluate currently available bisphosphonate drugs as a first line of defense for individuals exposed to F. tularensis; and attenuated, live-vector tularemia vaccine); 5) Low-Dose Enteric Pathogens (role of type 1 Cryptosporidium parvum candidate genes in pathogenesis/susceptibility to infection as a prelude to vaccine development; Shigella dysenteriae 1 and EHEC vaccines; novel therapeutics for EHEC disease; and diagnostics for detection of these pathogens in water, food, and environmental specimens); and 6) Public Health Response Research (needle-free immunization and vaccine-adjuvanting strategies; immunogenetics of human immune response to smallpox vaccine; and innovative diagnostic platforms for routine clinical use and in known or suspected bioterror events). Three Career Development Projects (to train the next generation of biodefense investigators) and four Developmental Projects (high-risk projects on biodefense agents) will be funded per year. Training will include a Media Training Course; a short course in "GMP Production and Process Development" (in collaboration with Aventis Pasteur Vaccines and Merck Vaccines); a Category A Bioterror Agent Clinical Surveillance Course; a "hands-on" course on working in BSL-3 facilities; and travel awards for RCE scientists and trainees to visit other RCE labs to learn techniques or perform collaborative experiments. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •
Project Title: DEVELOPMENT OF A SAFER SMALLPOX VACCINE Principal Investigator & Institution: Cho, Michael W.; Assistant Professor; Medicine; Case Western Reserve University 10900 Euclid Ave Cleveland, Oh 44106 Timing: Fiscal Year 2003; Project Start 30-SEP-2002; Project End 31-AUG-2004 Summary: (provided by applicant): Smallpox, which is caused by variola virus, is a highly contagious disease with a high fatality rate. A successful, worldwide vaccination campaign during the 1950s-l970s, using live vaccinia virus, resulted in eradication of smallpox. However, there remains a remote, but distinct possibility that large stockpiles of the virus may have been produced and stored as a part of bioweapons program in some countries or by terrorist organizations that are presently hostile to the United States. Smallpox poses a grave danger as an agent of biological weapon because of its highly contagious nature. Since vaccination against the disease stopped during early 1970s in the U.S., a large number of the young generation is unvaccinated and vulnerable to possible bioterrorist attacks. Although existing smallpox vaccine is relatively safe, it is not without serious medical complications including eczema vaccinatum, vaccinia necrosum, and encephalitis. Given the high frequency of AIDS patients who are immunodeficient, use of the current vaccine could result in a serious public health disaster. Therefore, it is imperative to consider developing a secondgeneration smallpox vaccine that is safer, yet as effective as the existing vaccine. Presently, variola virus is not available to perform additional research and the immune correlate of protection against either variola or vaccinia virus is largely unknown. Given these circumstances, a study is proposed with a long-term goal of developing a safer smallpox vaccine, with the following specific aims: (1) to characterize humoral and cellular immune responses against vaccinia virus in macaques previously immunized
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with the virus; (2) to evaluate immunogenicity and degree of attenuation of three recombinant vaccinia viruses derived from two different vaccinia strains (Western Reserve and Wyeth) in mice; and (3) to compare immunogenicity and safety of the newly generated vaccinia virus(es) with those of currently licensed smallpox vaccine strain in macaques previously infected with chimeric SlV/HIV -1 (SHIV). A successful completion of the proposed projects would allow better understanding of immune correlates of protection against vaccinia virus and could facilitate development of a safer smallpox vaccine. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •
Project Title: DEVELOPMENTAL RESEARCH PROJECTS Principal Investigator & Institution: Schlievert, Patrick M.; Professor; University of Minnesota Twin Cities 200 Oak Street Se Minneapolis, Mn 554552070 Timing: Fiscal Year 2003; Project Start 04-SEP-2003; Project End 31-AUG-2005 Summary: Ten projects were submitted by the applicant. The principal strengths identified by reviewers include: 1) the scientific merit and wide scope of research project topics and approaches, 2) a comprehensive timetable for the evaluation of productivity of the projects, 3) a reasonable plan for soliciting projects and awarding funds, 4) the relevance of most of the proposed projects to the overall program plans, 5) the potential of several projects to become full-fledged research projects for the RCE application, and 6) the highly qualified investigators. Principal weaknesses include: 1) the lack of detail regarding the selection, evaluation and management of primary research projects for the RCE application, 2) some developmental research projects, especially those involving viral agents, require strengthening, 3) the insufficient detail regarding the recruitment of women and under-represented minorities, and 4) lack of information about establishment of long-term research partnerships within and outside the region. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen
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Project Title: DISCOVERY OF SUBUNIT VACCINES FOR SMALLPOX Principal Investigator & Institution: Sykes, Kathryn F.; Director of Vaccine Research; Macrogenics, Inc. 1500 E Gude Dr, Ste B Rockville, Md 20850 Timing: Fiscal Year 2003; Project Start 04-SEP-2003; Project End 29-FEB-2008 Summary: Eradication of smallpox as a natural pathogen happened 20 years ago. Eradication of smallpox as a biothreat is now our objective. New stockpiles of safe and efficacious smallpox vaccine are needed to protect both the civilian and military populations against deliberate release of the smallpox virus. Currently there is no commercially available vaccine, and the previously approved one is a live vaccinia inoculum associated with more adverse events than any other approved vaccine. Recent work has focused on improving the manufacturing process of the original vaccinia vaccine strain and testing other live attenuated viruses. We propose to discover new vaccine candidates from viral components. A subunit design would be safer and more easily controlled during manufacture. Since we anticipate obtaining multiple protective subunits, these can be mixed and matched to effectively defend against wild type, natural variants, and bioengineered smallpox isolates. To identify antigens of smallpox that carry vaccine potential, the goal of our proposed project is to screen all the genes of the closely related cowpox virus for their ability to protect against disease in its natural murine host. This genome-level approach is feasible because the viral genome databases are available and we have developed the platform technologies that make a comprehensive screen fast and efficient. We will establish the electronic and molecular
12
Smallpox Vaccine
protocols to synthetically generate a thousand codon-optimized gene sequences. It will be used to produce a library of high quality cowpox subgenes. An advanced library screening method employing multiplex arrays will enable us to screen all the subgenes for protection in one experiment. Vaccine candidates will be confirmed and immune characterized. Both the cowpox candidates and their variola homologs will be formatted three ways and evaluated in immune and cowpox protection assays. This project will uncover new subunit vaccine candidates against variola and prepare them for final validation in a primate challenge experiment. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •
Project Title: DNA DENGUE/VACCINIA
VACCINE
DELIVERY
FOR
BIODEFENSE
WITH
Principal Investigator & Institution: King, Alan D.; Chief Scientific Officer; Cyto Pulse Sciences, Inc. 7513 Connelley Dr, Ste C Hanover, Md 21076 Timing: Fiscal Year 2003; Project Start 15-APR-2003; Project End 31-JAN-2004 Summary: (provided by applicant): The long-term project objective is the commercialization of a safe, effective, easy to use, and painless polynucleotide vaccine delivery system that can be used in polynucleotide vaccines for biodefense against NIAID Category A, B and C Pathogens. Polynucleotide vaccines are on the forefront of vaccine development. They are important because of the fast development times possible and because cell mediated immune responses can be induced. The delivery system proposed here will be effective for most polynucleotide vaccines. This delivery system specifically addresses the requirement as presented in the NIAID Strategic Plan for Biodefense Research, February 2002, page 8. In addition to Biodefense, this system will provide effective polynucleotide vaccine delivery for less lethal viruses, some cancers and some third world diseases. The defense and commercial applications are extensive. The polynucleotide vaccine delivery system described here uses a microneedle array with the polynucleotide coated right on the needle in the array. There are hundreds of needles each about 0.15 mm long. This array in inserted into the skin with the needle penetrating to about the basal lamina. After insertion the polynucleotide leaves the needle surface and an electric field is used to permeabilize dendritic and epithelial cell membranes to permit the polynucleotide to enter the cell. The system will be tested with the WRAIR/Cyto Pulse dengue DNA vaccine which will be used as a model for hemorrhagic fever viruses and the USAMRIID vaccinia DNA plasmid which is the primary vaccine for small pox. The specific aims of this project are to design and develop to FDA QSR Standards the vaccine delivery system prototype and to test the prototype in a human trial. This is a fast-track application. In Phase I, a system design will be completed including the hand-piece, microneedle array and miniature waveform generator. The coating chemistry and specific waveforms will be optimized in mice. In Phase II, a prototype of the final design will be completed. Safety and efficacy will be demonstrated in mice and safety will be demonstrated in humans. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •
Project Title: DOMINANCE VACCINATION
IN
T
CELL
RESPONSES
TO
SMALLPOX
Principal Investigator & Institution: Sette, Alessandro B.; Head and Member; La Jolla Institute for Allergy/Immunolgy 10355 Science Center Dr San Diego, Ca 921211118 Timing: Fiscal Year 2003; Project Start 01-JUL-2003; Project End 31-DEC-2006
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Summary: (provided by applicant): Vaccination against variola virus (the causative agent of smallpox) is currently accomplished by vaccinia virus. Little is known about 1) the antigens and epitopes targeted by the cellular responses in humans immunized with vaccinia virus, and 2) which responses are crossreactive with variola virus and hence would be expected to contribute to the protection engendered by the vaccine. In the first part of the studies proposed herein, we will 1) determine immunodominant antigens recognized by Class I and Class II restricted responses in humans immunized with vaccinia virus, 2) map the epitopes recognized within each antigen, and 3) determine their degree of crossreactivity with homologous variola virus-derived sequences. We anticipate that these studies will lead to the definition of a broad range of epitopes, facilitate a rigorous definition of correlates of protection against smallpox infection in humans, and also enable the evaluation of the performance of different vaccine candidates. Vaccinia virus is also actively investigated as a potential vaccine delivery vehicle, either alone or in prime/boost regimens, for disease indications such as HIV, malaria and cancer. Thus, it should be noted that identification and characterization of the determinants recognized by humans infected/vaccinated by vaccinia virus would also enable the characterization and optimization of experimental vaccines utilizing vaccinia virus-derived vectors as a delivery system. The vaccinia-based vaccines currently available, while effective, are associated with significant and serious, albeit rare, side effects. Because of these side effects, and because of the worldwide eradication of variola virus, vaccination of the general population was deemed as no longer desirable. Recent renewed concerns have been raised over bioterrorist use of the virus. In the context of the studies proposed herein, a concern could be raised that if the vaccinia-induced protection is mediated by relatively few immunodominant and crossreactive antigens, a modified smallpox virus could be engineered that lacks those crossreactive epitopes. Under this terrifying scenario, the protection elicited by the vaccinia would be ineffective against the biological weapon. In the second part of the grant, we propose to counter this risk through the identification of variola virus-specific determinants derived from immunodominant antigens in the context of the vaccinia virus responses, but not crossreactive with the homologous variola virus sequences. These variola virus-derived epitopes would be incorporated in an optimized multideterminant vaccine construct, inserted in the currently available vaccinia vaccine. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •
Project Title: HETEROPOLYMER COMPLICATIONS
SYSTEM
TO
TREAT
VACCINIA
Principal Investigator & Institution: Pincus, Steven E.; Elusys Therapeutics 10 Bloomfield Ave Pine Brook, Nj 07058 Timing: Fiscal Year 2003; Project Start 15-MAR-2003; Project End 28-FEB-2005 Summary: (provided by applicant): Smallpox is a particularly dangerous biological weapon because it can be manufactured in large quantities, stored for an extended period of time, and delivered as an infectious aerosol. It is highly infectious and has a death rate as high as 25%. The only approved smallpox vaccine (live vaccinia virus) is available in very limited quantities (15.4 million doses) and is decades old. Current efforts are underway to produce enough vaccine to vaccinate the entire US population. A problem with this vaccination strategy is that a large segment of the population is susceptible to severe adverse reactions associated with the vaccine, including bloodborne dissemination of the vaccinia virus and even death. This threat is even greater now than when smallpox vaccinations were routine (prior to 1974) due to the growing population of immunosuppressed individuals. The broad long-term objective
14
Smallpox Vaccine
of the project is to develop bispecific antibodies (Heteropolymers, HPs) for treatment of complications associated with the administration of smallpox vaccine. In the present work, we hypothesize that vaccinia virus, the active component of smallpox vaccine, can be bound to erythrocytes (Es) via HPs, cleared to acceptor macrophages, and destroyed without killing the target cells. The HPs will consist of one monoclonal antibody (MAb) against a vaccinia protein expressed on the surface of extracellular (EEV) or intracellular (IMV) virus, cross-linked to a second MAb specific for E Complement Receptor Type I (CR1). The goal of the present work is to identify at least one anti-vaccinia HP that will be a candidate for use in future primate studies to establish efficacy against vaccinia complications following administration to immunocompromised animals. To achieve that goal, we will screen the anti-vaccinia MAb panel to identify the high affinity antibodies that bind EEV or IMV forms in solution phase and, preferably, neutralize the virus. The selected MAbs will be used to prepare HPs by cross-Iinking to a MAb specific for human E CR1. The ability of these HPs to bind EEV or IMV forms of vaccinia and transfer it to acceptor macrophages will be tested. We will also determine whether HP bound vaccinia is infectious. We will determine whether these HPs can prevent vaccinia virus spread and pathology in immunocompromised transgenic mice (expressing human CR1). Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •
Project Title: HIGH TITER VIG FOR THE TREATMENT OF SMALLPOX Principal Investigator & Institution: Nur, Israel; Omrix Biopharmaceuticals Science Park Bldg 14 Nes-Ziona, Timing: Fiscal Year 2003; Project Start 19-SEP-2003; Project End 31-AUG-2005 Summary: (provided by applicant): Long-term objectives Based on preliminary studies on Israeli volunteers, it was concluded that only 60% of the population revaccinated against Smallpox reacted to the vaccine, of whom only 10% did so at very high titers. The aim of this R&D project is to develop and produce a high-titer, small volume Vaccinia Immune Globulin (HT-VIG) as an effective countermeasure to Smallpox and the side effects of the Vaccinia vaccine, which can be administered by intra-muscular or intra-venous mutes, including self-injection. The resulting product will be at least ten times more concentrated than existing VIG preparations that are based on immunoglobulin preparations from non-selected, pooled plasma, and it will have greater efficacy. Importantly, it will be a safe and effective prophylactic treatment against Smallpox for people excluded from the Vaccinia vaccination due to immunodeficiency or other risk factors. Given the heightened risk of a Smallpox outbreak due to bio-terrorism, the project's importance and health relevance cannot be underestimated. Specific Aims Omrix' existing ELISA test will be validated, establishing the correlation between it and the standard Vaccinia neutralization assay. The validated ELISA test will be used to determine the level of anti-Vaccinia immunoglobulins in human plasma samples. Following that, the ELISA will be used to screen USA plasma derived from re-vaccinated donors and select high titer anti Vaccinia plasma samples. These samples will be used to produce a limited number of batches of High-Titer VIG, which will then be characterized by use of the ELISA test and the standard Vaccinia Neutralization Bioassay. Development of a concentrated HT-VIG formulation for either small volume IV or IM administration. Research Design & Methods As well as employing standard operating procedures for plasma collection and fractionation, innovative methodology will be employed in two areas of the program: 1) a new validated ELISA screening test will enable selection of very high titer plasma samples resulting in a high potency, small volume HT-VIG product; 2) the development of the
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HT-VIG product will include the new viral removal steps developed by Omrix, such as nano-filtration and solvent detergent treatment, which exhibit high margins of viral inactivation of all known viruses, including Parvo-viruses. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •
Project Title: HUMAN IMMUNITY TO VACCINIA VIRUS Principal Investigator & Institution: Kazura, James W.; Professor; Case Western Reserve University 10900 Euclid Ave Cleveland, Oh 44106 Timing: Fiscal Year 2003; Project Start 01-AUG-2003; Project End 31-JUL-2008 Summary: Current policy to protect Americans against smallpox mandates immunization of naive and previously vaccinated adults with live vaccinla virus (VV). Knowledge of adaptive immunity to VV is limited to descriptions of neutralizing antibodies, cytotoxic T lymphocyte (CTL) activity, and lymphocyte proliferation responses, largely because widespread vaccination ceased before the current era of molecular immunology. Research in this project will examine the evolution of VVspecific memory T cells in newly vaccinated and re-vaccinated (boosted) adults. The relationship of VV-specific adaptive immunity to viral load and innate immunity will be determined in conjunction with projects by Storch and Stanley. The specific aims are to: 1. Identify immunodominant VV epitopes that induce CD8+ T cell memory. Artificial neural networks and computational algorithms will initially be used to select peptide epitopes, followed by evaluation of peptide binding to HLA class I alleles and assessment of the ability of peptides to stimulate IFN-gamma and IL-2 responses by HLA-matched and HLA-mismatched primary vaccinees and adults given booster doses of VV. 2. Determine the functional phenotype of CD8+ cells during the inductive and memory phases of adaptive immunity to VV. HLA class I tetramer-peptide complexes will be used to identify and quantify CD8+ cells bearing TCR specific for VV epitopes before vaccination and 9 days, 6, 12, and 24 months post-vaccination. Lymphocytes producing putative mediators of immunity (IFN-gamma, perforin, MIP-1alpha) and bearing memory markers (CD45RO, CD45RA, CCR7) will be evaluated prospectively in primary vaccinees and boosted adults. 3. Evaluate the clonotypic repertoire of VVspecific CD8+ cells following primary vaccination and boosting of previously vaccinated adults. TCR Vbeta usage and CDR3 length and sequence polymorphism will be evaluated prospectively in selected HLA-A2+ and HLA-A11+ individuals. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen
•
Project Title: HUMAN MONOCLONAL ANTIBODIES TO REPLACE VIG FOR THERAPY Principal Investigator & Institution: Cavacini, Lisa A.; Assistant Professor of Medicine; Beth Israel Deaconess Medical Center St 1005 Boston, Ma 02215 Timing: Fiscal Year 2003; Project Start 27-SEP-2002; Project End 31-AUG-2004 Summary: (provided by applicant): Prior to Edward Jenner's demonstration in 1796 that immunization with cowpox protected against smallpox infection, virtually everyone contracted smallpox with mortality as high or higher than 30%. A global campaign using vaccinia immunization for protection from smallpox infection resulted in the eradication of smallpox in 1977. Subsequently the WHO recommended that all countries cease vaccination and laboratory stocks be destroyed or transferred to one of two repositories at the CDC in the United States or the Institute of Virus Preparations in Moscow, Russia. Routine vaccination in the United States has not occurred for more than thirty years. Therefore, the vast majority of the population is at risk of smallpox
16
Smallpox Vaccine
infection. Just as smallpox was used as a bioweapon prior to the development of vaccination, it currently represents a potential biological weapon with the majority of the world population at risk. Vaccination within the first few days after exposure is effective at preventing infection in some with a significant decrease in mortality. However, there are rare and serious complications in some vaccinated individuals. The vaccination is contraindicated in a number of groups of people. Vaccinia immune globulin (VIG) has been used as prophylaxis for treating individuals for which contra indications exist for smallpox vaccine and for treatment of those with complications of vaccinations. With the threat of smallpox being used as an agent of bioterrorism, it is prudent to develop alternatives for the use of VIG for prophylaxis and treatment. Supplies of VIG are scarce given that individuals have not been systematically vaccinated for more than 30 years. Furthermore, the validation and safety of VIG remains an issue. Therefore, we propose to generate human monoclonal antibodies as a replacement for VIG. Monoclonal antibodies can be produced with exquisite specificity and can be modified to enhance functional activity. The use of fully human monoclonal antibodies eliminates problems associated with xenogeneic, chimeric or humanized antibodies which include immunogenicity, biological half-life, and inefficient effector function. The development of a cocktail of human monoclonal antibodies that neutralize virus and/or mediate antibody-dependent cellular cytotoxicity can serve as a safe, effective replacement for VIG. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •
Project Title: HUMAN T CELL RESPONSES TO VACCINIA VACCINE Principal Investigator & Institution: Flomenberg, Phyllis R.; Medicine; Thomas Jefferson University Office of Research Administration Philadelphia, Pa 191075587 Timing: Fiscal Year 2004; Project Start 15-APR-2004; Project End 31-MAR-2006 Summary: (provided by applicant): There is an urgent need to develop safer smallpox vaccines. One approach is the use of modified vaccinia virus Ankara (MVA), a highly attenuated vaccinia virus that does not replicate in human cells. In comparison to vaccinia virus, however, MVA is less immunogenic and may require higher doses and more frequent boosts. There also remain safety concerns about the use of a live attenuated virus and the potential presence of adventitious pathogens. Alternative approaches include the development of inactivated virus or recombinant protein vaccines. However, vaccine development is hampered because little is known about what proteins play an important role in the generation of protective immune responses to smallpox. Recently, 4 envelope proteins that induce neutralizing antibodies were identified, and DNA vaccination with a combination of all 4 genes protected mice from a lethal vaccinia infection. Although a vaccine will need to induce strong T cell responses, in addition to neutralizing antibodies, no information is available about the vaccinia protein targets recognized by T cells. In particular, CD4 T cells are required to enhance B cell, T cell, and innate immune responses to lytic viruses. As a first step, peripheral blood mononuclear cells from healthy adults previously immunized with vaccinia virus will be tested for virus-specific T cell responses. The goals of this study are: 1) to measure the frequencies of human T cell responses to vaccinia structural proteins; 2) to identify major CD4 and CD8 T cell epitopes within the above 4 envelope proteins; and 3) to analyze the frequencies of the T cell responses to these epitopes postvaccination. It is anticipated that this work will provide new tools for the detection and monitoring of vaccinia-specific T cell responses from most individuals and will help develop new strategies for smallpox vaccine development. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen
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17
Project Title: IMMUNODOMINANT EPITOPES OF A SMALLPOX VACCINE IN HUMANS Principal Investigator & Institution: Buller, Mark R.; Professor; Molecular Microbiol and Immun; St. Louis University St. Louis, Mo 63110 Timing: Fiscal Year 2003; Project Start 01-SEP-2002; Project End 31-AUG-2004 Summary: (provided by applicant): The zenith of the disease smallpox and its eradication in 1977 from human populations occurred prior to the modern era of immunology and molecular biology. Consequently there is little knowledge concerning the immune correlates for recovery from smallpox or the cross-reactive proteins expressed by baccinia virus that were responsible for its success as the smallpox vaccine. The only vaccination indicator that correlated with protection from severe smallpox was the scar. In response to the threat of bioterrorism, the U.S. government has redoubled its efforts to provide strategies that will protect the American public from an outbreak of smallpox or human monkeypox. As part of a comprehensive, multi-faceted plan, the U.S. government has contracted with Acambis Inc. and Baxter Healthcare Corp. to produce approximately 209 million doses of a new tissue culture smallpox vaccine. In addition, proposals are being considered for the next generation of smallpox vaccine that will have an enhanced safety profile, causing fewer vaccine-related complications, especially in immunosuppressed individuals. Evaluating the efficacy of the new Acambis and Baxter vaccine or other second-generation vaccines with enhanced safety profiles will be problematic without detailed knowledge of the immunogenicity of vaccines proven to be efficacious in the smallpox eradication program. Detailed studies on human B and T cell immune responses to proteins encoded by vaccinia virus should help fill this gap in knowledge, and may also identify targets of neutralizing, complement-fixation or ADCC (antibody-dependent cell cytotoxity) antibodies, which may facilitate the development of an efficacious replacement for VIG. We propose to characterize the vaccinia virus-encoded proteins recognized by B and T cell responses during the vaccination of volunteers with the DryVax vaccine. The Specific Aims are to: 1. Characterize the antibody responses to immunodominant vaccinia virus proteins and 2. Identify the epitope specificity of representative vaccinia virus-specific T cell clones. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen
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Project Title: IMMUNOGENETICS OF SMALLPOX VACCINATION Principal Investigator & Institution: Stanley, Samuel L.; Professor; Washington University Lindell and Skinker Blvd St. Louis, Mo 63130 Timing: Fiscal Year 2003; Project Start 01-AUG-2003; Project End 31-JUL-2008 Summary: The goal of this project is to identify genes that are involved in susceptibility and resistance to human vaccinia infection, and, consequently, in some of the adverse effects seen with smallpox vaccination. We (R.B.B.) recently led a multi-center prospective study on the clinical response to vaccinia immunization in 680 naive individuals. Among the 665 individuals responding to the vaccine, 84 (13%) developed fever, muscle aches and lymphadenopathy giving rise to what we have called Acute Vaccinia Syndrome (AVS) in approximately 30% of vaccines. The timing of the onset of these symptoms matched the timing of the highest levels of viral shedding, indicating that fever, and the other components of AVS appear to be secondary to the virus. We hypothesize that individuals developing AVS (and especially fever) have diseasepredisposing alleles that are associated with abnormal innate immune or delayed adaptive immune responses to vaccinia. These individuals may be more susceptible to poxviruses in general, and could constitute a group at increased risk for mortality if
18
Smallpox Vaccine
exposed to smallpox. We propose to identify genes that are expressed in response to vaccinia infection at the site of inoculation and systemically using a transcriptional analysis. We will compare responses between individuals that develop AVS, and those individuals who develop no adverse reactions to immunization. These studies will provide us with a transcriptional profile of the host response to vaccinia infection, identify key molecules in the host response, and, establish parameters for protective immune responses that could be used to test the efficacy of new vaccines. We will also identify alleles associated with adverse effects to vaccinia immunization and abnormal innate immune responses to the virus through the analysis of haplotypes based on single nucleotide polymorphisms in candidate genes. The identify of these alleles may provide clues to the critical elements of the host response to poxvirus, and could provide a method to identify individuals at increased risk for adverse effects to the vaccine, or more severe disease with poxvirus infection. The design of the study, with the inclusion of transcriptional profiling of individuals receiving vaccinia immunization coupled with a detailed virologic and immunologic profile, ensures that we will obtain valuable information on the host response to vaccinia immunization. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •
Project Title: NORTHEAST BIODEFENSE CENTER Principal Investigator & Institution: Lipkin, W Ian.; Professor; Wadsworth Center Empire State Plaza Albany, Ny 12237 Timing: Fiscal Year 2003; Project Start 04-SEP-2003; Project End 29-FEB-2008 Summary: (provided by applicant): The northeastern United States is highly vulnerable to emerging infectious diseases and terrorism. With the outbreak of West Nile virus, the World Trade Center tragedy, and subsequent anthrax attacks, Region II has the experience and resolve to mobilize its rich resources in biomedical research to advance the nation's biodefense agenda. Region II institutions will establish a Regional Center of Excellence (RCE) for Biodefense and Emerging Infectious Disease Research. This consortium, the Northeast Biodefense Center (NBC), will comprise investigators at more than 25 academic and research institutions in New York, New Jersey, Connecticut, Massachusetts and Puerto Rico. The NBC will have strong links to state, federal and local government agencies and laboratories, as well as biotech and pharmaceutical companies. Major basic and translational research programs will be pursued in six thematic areas: 1) B-Cell Related Prophylaxis and Therapeutics; 2) Bacterial Pathogenesis and Therapeutics; 3) Vaccine Platforms; 4) Viral Pathogenesis and Therapeutics; 5) Smallpox Vaccine: Clinical, Immune, and Viral Outcomes; 6) Pathogen Detection and Diagnostics. These programs will intersect with cores that provide support in Informatics, Proteomics, Protein Expression, Monoclonal Antibodies, Animal Models, as well as Administration. Each research program will integrate and intensify the work of several accomplished investigators and will focus on select agents and toxins including Bacillus anthracis, Yersinia pestis, Francisella tularensis, Burkholderia mallei, Staphylococcus enterotoxin B, pox viruses, and viruses that cause hemorrhagic fever or encephalitis. Zoonotic diseases will be emphasized due to significance in this region, expertise of NBC members, and access to unique resources such as Plum Island Animal Disease Center and networks of investigators and clinicians in comparative medicine. Streamlined technology transfer procedures will be established to facilitate delivery to industry of vaccines, therapeutics, and diagnostics. The translational arm of the NBC has begun with a study of smallpox vaccination and will develop the region's infrastructure for human vaccine trials. Training programs will be established to promote biodefense research objectives by supporting new investigators, senior
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investigators, and support personnel. An emergency response plan has been developed to rapidly realign the NBC's activities and provide facilities, including cores and scientific support, to first line responders in the event of a biodefense emergency. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •
Project Title: NOVEL VACCINES FOR SMALLPOX Principal Investigator & Institution: Weiner, David B.; Associate Professor of Pathology and Lab; Medicine; University of Pennsylvania 3451 Walnut Street Philadelphia, Pa 19104 Timing: Fiscal Year 2003; Project Start 15-MAY-2003; Project End 30-APR-2008 Summary: (provided by applicant): The current bioterrorism threat has refocused our nation on the issue of our population's susceptibility to a smallpox attack. As a precaution, deployment of dilution of the current stock of vaccine and deployment of a tissue culture version of the VACV vaccine have been requisitioned. This vaccine and likely the new stocks carried a risk of Significant Adverse Events of 1 per 10,000 vaccinated individuals. However, that vaccine was last used in a very different environment. The high-risk groups for AE's include the elderly, the very young, immunocompromised individuals and others. The percentage and absolute numbers of the US population that falls within these categories has risen dramatically in the past 30 years. The greater than 750,000 persons living in the US that are HIV-positive is one clear example. This suggests that the AE risk of the VACV may be unacceptable and can only be deployed as a last resort. However, the option of abandoning this approach and developing new approaches leaves us at risk for a possibly unknown period of time. The hypothesis to be tested in this application is that there is a third option, to develop a strategy that uses the current vaccine yet limits its pathogenesis while improving its potency. It is our hypothesis that priming with enhanced expressing plasmid vaccines that induce nonneutralizing cellular immune responses will prime for successful and even enhanced boosting with the current vaccine, yet limit its associated pathogenesis. This application will use quantitative T cell assays including Elispot and ICC and tetramer analysis, and novel human HLA+DR positive transgenic mice that we have developed to pursue the three specific aims. Novel cellular reagents including MHC class I tetramers will be developed that will have significant value in following VACV challenge in humans as they may be useful as cellular surrogates for the current site reaction take. Together these studies will establish if this simple and novel approach can bridge the current situation and produce a safer more effective smallpox vaccine. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen
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Project Title: ORTHOPOXVIRUS VACCINE DEVELOPMENT Principal Investigator & Institution: Feinberg, Mark A.; Associate Professor; Emory University 1784 North Decatur Road Atlanta, Ga 30322 Timing: Fiscal Year 2003; Project Start 01-AUG-2003; Project End 31-JUL-2008 Summary: The long-term objectives of this project are to produce novel vaccines and immunotherapeutics designed to abrogate the activities of poxvirus-encoded extracellular accessory proteins (XAPs). We will focus on the XAPs that interfere with host responses, particularly immune responses, against poxvirus infections. We will test the hypothesis that preexisting immunity to the XAPs will abrogate the in vivo replicative advantages that poxviruses gain from expressing these XAPs, and reduce viral virulence in the immunized host. The specific aims of this project are as follows: 1. Construct Venezuelan Equine Encephalitis (VEE) virus replicons expressing mutationally
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Smallpox Vaccine
inactivated XAPs of cowpox virus (CPXV). Initially, we will focus on the extra-cellular viral inhibitors of complement and interferon. Subsequently, we will target the other orthopoxvirus XAPs, each of which is encoded by CPXV. 2. Assemble and test prototype VEE replicon particle (VRP) vaccines for CPXV-XAPs: VRPs will be assembled containing replicons expressing the genetically inactivated XAPs. We will determine the immunological responses of mice vaccinated with the VRP-CPXV-XAP vaccines. We will determine the abilities of antisera from immunized mice to neutralize the in vitro activities of the target XAPs, as well as cross-protective properties of the VRP-CPXVXAP-induced antibodies against accessory proteins encoded by other orthopoxviruses, including variola virus. 3. Evaluate the efficacy of prophylactic VRP-CPXV-XAP vaccines in the mouse model: We will determine if vaccination with the VRP-CPXVXAP vaccines can reduce CPXV replication and pathogenesis following challenge with CPXV in the mouse. We will also examine the effects of these VRP vaccines on infections with vaccinia viruses, and we will determine the efficacy of prime boost regimens involving primary immunization with VRP-CPXV-XAP vaccines followed by vaccination with vaccinia virus vaccines. 4. Evaluate the efficacy of therapeutic VRPCPXV-XAP vaccines and anti-XAP antibodies in the mouse model: We will determine if post-infection vaccination with VRP-CPXV-XAP vaccines or post-infection treatment with monoclonal antibodies against XAPs encoded by cowpox virus can attenuate cowpox and vaccinia virus infections in the mouse. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •
Project Title: POX VIRUS IMMUNOLOGY AND VACCINE DEVELOPMENT Principal Investigator & Institution: Ahmed, Rafi; Director; Microbiology and Immunology; Emory University 1784 North Decatur Road Atlanta, Ga 30322 Timing: Fiscal Year 2003; Project Start 15-SEP-2003; Project End 31-DEC-2007 Summary: (provided by applicant): There is a serious need for a smallpox vaccine alternative because of the significant incidence of adverse events to the current vaccine (Dryvax). Large groups in the American population are not qualified to receive the current smallpox vaccine due to immunodeficiency (from genetic causes, HIV, or immunosuppressive drugs), old age, skin disorders, young age (< 1 yr), or pregnancy. These groups are major populations and must be accounted for in any reasonable national smallpox vaccination strategy. Therefore, assessment of the immunogenicity of alternative smallpox vaccines such as modified vaccinia Ankara (MVA) must be done accurately and expeditiously. Our access to samples from ongoing clinical trials of Dryvax and MVA, our expertise in quantitating both cellular and humoral immunity, and our immunologic proteomics approach places our research group in a unique position to address this important issue. Long term protective immunity to smallpox is likely provided by three arms of the immune system: circulating neutralizing antibodies, memory B cells, and memory T cells. Virtually none of the vaccinia protein targets responsible for these T cell, B cell, and neutralizing antibody responses have been identified. The experiments in this proposal are designed to determine the dominant anti-smallpox immune responses and to compare the magnitude and breadth of the T and B cell responses induced by Dryvax versus MVA. This valuable information will not only allow an accurate assessment of the quality of the immune responses elicited by the alternative smallpox vaccine MVA but can also then be immediately parlayed into additional areas of proposed research such as: 1) diagnostic tools for measuring long term smallpox immunity in vaccinees; 2) development of simple, rapid, and sensitive immunological tools for assessing recent smallpox (variola major) exposure/infection; and 3) development of therapeutic anti-smallpox neutralizing monoclonal human
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antibodies. Finally, the knowledge acquired from the detailed studies described herein regarding the immunodominant protein targets of human vaccinia-specific T and B cells will lay the foundation for developing a safe and effective smallpox subunit vaccine. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •
Project Title: POXVIRUS ADVERSE EFFECTS ON CARDIAC CELLS AND THE HEART Principal Investigator & Institution: Sherry, Barbara A.; Professor; Molecular Biomedical Sciences; North Carolina State University Raleigh 2230 Stinson Drive Raleigh, Nc 27695 Timing: Fiscal Year 2004; Project Start 01-APR-2004; Project End 31-MAR-2006 Summary: (provided by applicant): In response to concerns that bioterrorists may release smallpox virus in the United States, the government has implemented a multistage vaccination program. However, the observation of cardiac adverse events following vaccination has resulted in the Centers for Disease Control recommending that people at risk for heart disease should be excluded from the vaccination program. Notably, myocarditis has been diagnosed in 18 recent smallpox vaccinees in the USA. Despite this recent temporal association between myocarditis and smallpox vaccination as well as a similar historic association, there have been no studies of the effects of poxviruses on cardiac cells, or the development of any animal models to study poxvirus-induced myocarditis. For other viruses, myocarditis can reflect a direct cytopathogenic effect of the virus on cardiac myocytes, or can be mediated by immune cells, cytokines, or other soluble factors. Poxviruses can be directly cytopathogenic, but also encode a number of modulators of cytokine responses. Cowpox virus contains the full complement of known orthopoxvirus accessory genes that affect immune responses, while DryVax (the current vaccine strain of vaccinia virus) encodes fewer accessory genes, and modified vaccinia Ankara virus (MVA, which is replication-defective in human cells) contains even fewer accessory genes. Our preliminary data demonstrate that cowpox virus and DryVax replicate in murine primary cardiac myocyte cultures (PCMCs) whereas MVA does not. Moreover, cowpox, DryVax, and MVA are each cytopathogenic to these cultures, though the type of cytopathogenicity is virus strainspecific. We hypothesize that poxviruses have virus strain-specific adverse effects on cardiac cells, either directly or indirectly, resulting in damage to cardiac myocytes and the heart. In Specific Aim 1, we will determine the effect of these three poxviruses (cowpox, DryVax, and MVA) on murine primary cardiac myocyte cultures (PCMCs) and control cultures, analyzing viral infection and gene expression, and induction of cytopathogenicity. In Specific Aim 2, we will determine the association between cytokines and poxvirus infection on cytopathogenicity to PCMCs and control cultures. In Specific Aim 3, we will determine whether these poxviruses induce myocarditis in a variety of mouse strains. Studies proposed in this R21 application will provide the foundation for future in-depth studies with the goal of Improving current and future smallpox vaccines. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen
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Project Title: POXVIRUS IMMUNITY AND DNA/MVA HIV VACCINES Principal Investigator & Institution: Amara, Rama R.; None; Emory University 1784 North Decatur Road Atlanta, Ga 30322 Timing: Fiscal Year 2003; Project Start 01-JUL-2003; Project End 31-OCT-2007 Summary: (provided by applicant): We recently demonstrated the ability of an AIDS vaccine consisting of DNA priming and recombinant modified vaccinia Ankara (MVA)
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booster immunizations (DNA/MVA SHIV vaccine) to control a pathogenic SHIV 89.6P challenge that was administered seven months after the final immunization in macaques. The prototype HIV-1 clade B version of our DNA/MVA vaccine (DNA/MVA HIV vaccine) is entering phase I safety trials in humans in January of 2003. Due to the recent bioterrorism threat the US government is prepared to vaccinate at least a subset of people with the current smallpox vaccine (Dryvax/New York Board of Health strain of vaccinia). The anti-vaccinia virus immunity generated by Dryvax may limit the boosting ability of MVA, hence the efficacy of DNA/MVA HIV vaccines. This is a very important question that needs to be addressed as DNA/MVA vaccines go forward in human trials. There is a serious need for a smallpox vaccine alternative because of the high incidence of adverse events to the current vaccine. Also, many people are not qualified to receive the current smallpox vaccine due to immunodeficiency, skin disorders, old age, young age (< 1 yr), or pregnancy. These groups are major populations and must be accounted for in any reasonable national smallpox vaccination strategy. MVA was developed towards the end of smallpox eradication for use in immunocompromised individuals and was used to vaccinate about 120,000 individuals. However, because smallpox had been controlled in first world countries by the time that MVA was developed, individuals who were vaccinated with MVA were not exposed to variola, and the efficacy of MVA as a smallpox vaccine was not determined. In this proposal we wish to address 1) the effect of preexisting immunity to smallpox on the ability of DNA/MVA vaccine to control pathogenic SHIV challenge, 2) the ability of vaccinia-specific immune responses raised by DNA/MVA vaccine to protect from a lethal monkeypox challenge and 3) the ability of a candidate DNA/MVA vaccine to control both SHIV and monkeypox challenges that are administered sequentially in the presence and absence of preexisting immunity to smallpox. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •
Project Title: POXVIRUS VACCINE RESEARCH Principal Investigator & Institution: Isaacs, Stuart N.; Assistant Professor; University of Pennsylvania 3451 Walnut Street Philadelphia, Pa 19104 Timing: Fiscal Year 2003; Project Start 01-AUG-2003; Project End 31-JUL-2008 Summary: Bioterrorism with variola virus is of immense concern because (a) virtually the entire world population is susceptible since routine vaccination was discontinued; (b) there are no treatments; (c) the virus in aerosol form is stable; (d) the virus is transmissible person-to-person; and (e) infection results in high morbidity and mortality. Vaccination with vaccinia virus (VV) was a key factor in eradicating smallpox. The necessity to vaccinate an at-risk population with W is central to preparing for the potential threat of smallpox bioterrorism. However recognized complications of vaccinia vaccination, especially in immunocompromised hosts, pregnant women, and infants impose serious limitations of this strategy. In past vaccination efforts, such complications were treated in the U.S. with human vaccinia immune globulin (VIG) obtained from W immunized people. Current stocks of VIG are low, and while new stocks are being generated, there are still serious drawbacks to relying on a blood product. Consequently, there is a critical need to develop therapeutic interventions to counter complications from the current vaccine and to develop a safer vaccine. As part of the mid-Atlantic Regional Center of Excellence in Biodefense & Emerging Infectious Diseases, our poxvirus research project's hypothesis is that vaccine candidates and new therapies can be developed by understanding and targeting poxvirus proteins recognized by the humoral and innate immune system. To do this we will: 1. Develop a
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subunit vaccine against smallpox (variola) virus (Cohen/Eisenberg/Friedman, U. Penn) 2. Identify new targets of neutralizing antibody (Isaacs, U. Penn) 3. Identify the targets of VIG using a proteomics approach (Lambris, U. Penn) 4. Develop an ectromelia virus challenge system in the mouse as a model of smallpox pathogenesis and prevention (Braciale, U. Virginia) Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •
Project Title: POXVIRUSES: DEFINING EPITOPE IMMUNODOMINANCE Principal Investigator & Institution: Terajima, Masanori; Univ of Massachusetts Med Sch Worcester Office of Research Funding Worcester, Ma 01655 Timing: Fiscal Year 2003; Project Start 01-OCT-2003; Project End 30-SEP-2008 Summary: Immunization with vaccinia virus resulted in long-lasting protection against smallpox and was the approach used to eliminate natural smallpox infections worldwide. This was accomplished without a detailed understanding of human T cell responses to poxviruses. Due to concerns about the use of smallpox virus as a bioweapon, smallpox vaccination is currently being reintroduced. Considering the relatively high incidence of side effects, developing a safer, but effective vaccine is very important. Vaccinia virus elicits strong cellular as well as humoral immune responses. Cellular immunity seems to be more important for recovery from infection. Severe complications from vaccination were associated with congenital or acquired T cell deficiencies, but not with congenital agammaglobulinemia. The presence of neutralizing antibody alone did not prevent the development of progressive vaccinia if cell-mediated immunity was defective. In order to analyze human T cell responses to licensed and experimental smallpox vaccines at the single cell level, it is essential to identify T cell epitopes, especially immunodominant CD8 + T cell epitopes. Vaccinia is a large virus and we hypothesize that we can develop a rapid approach to localize gene fragments encoding human T cell epitopes and to identify them precisely using peptides using PCR-generated DNA fragments containing virus genes transfected into antigen presenting cells. Complementary strategies will employ peptide libraries and studies in transgenic mice expressing common human MHC class I molecules. The immunodominance of human T cell epitopes on vaccinia virus will be analyzed. The results of this research project will provide valuable information relative to basic studies of human T cell memory and data useful for the design and analyses of experimental smallpox vaccines. The methods we will establish in this project may be applicable to other large viruses as well as bacteria for the definition of human T cell epitopes. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen
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Project Title: PUBLIC HEALTH RESPONSE Principal Investigator & Institution: Burke, Donald S.; Professor; Johns Hopkins University 3400 N Charles St Baltimore, Md 21218 Timing: Fiscal Year 2003; Project Start 04-SEP-2003; Project End 28-FEB-2008 Summary: This "Public Health Response Research" project is submitted as one of six research projects in the application for a multi-institutional Middle Atlantic Regional Center of Excellence (RCE) for Biodefense and Emerging Infectious Diseases Research. The objective of this project is to immediately accelerate the clinical evaluation and deployment of new biomedical technologies that may be crucial for an effective public health 0response to a bioterrorism or emerging infectious disease event. The project encompasses three sub-projects, all of which involve immediate evaluation of novel biotechnologies in human subjects. Sub-project 1 is designed to rapidly transition novel
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needle-free technologies for vaccine administration into routine use. The specific aims of this sub-project are to evaluate cutaneously administered immunostimulatory patches to boost responses to anthrax vaccines and botulinum toxoid, to evaluate intranasal administration of recombinant anthrax protective antigen, and to evaluate jet injector administration of recombinant anthrax protective antigen. Sub-project 2 is designed to use measurements of peripheral blood lymphocyte gene expression profiles from vaccinated persons to improve the diagnosis, prognosis, and prevention of smallpox vaccine adverse events. The specific aims of this sub-project are to define the natural history of vaccinia-induced immune dysregulation, to correlate favorable and unfavorable outcomes with specific gene transcript profiles, and to use the technology to identify individuals at increased risk of vaccine adverse reactions. Sub-project 3 is designed to evaluate rapid innovative genomics- and proteomics-based diagnostic technologies in real emergency room settings. The specific aims of this project are to evaluate novel universal bacterial and viral diagnostics, to evaluate blood lymphocyte gene expression profiles in common febrile illnesses, like influenza, that could be mistaken for bioterror threats, and to use mass spectrometry-based proteomics on respired air from patients to diagnose acute pulmonary infections. As additional new genomics- and proteomics-based technologies are generated from the Middle Atlantic RCE and elsewhere, they will be rapidly evaluated for their utility in the public heath biodefense response. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •
Project Title: SMALLPOX VACCINE CLINICAL RESEARCH CENTER Principal Investigator & Institution: Belshe, Robert B.; Professor of Medicine & Microbiology; Washington University Lindell and Skinker Blvd St. Louis, Mo 63130 Timing: Fiscal Year 2003; Project Start 01-AUG-2003; Project End 31-JUL-2008 Summary: With the imminent institution of smallpox vaccination for healthcare workers and first responders, there is an urgent need to better understand the biology of vaccinia infection in humans. In response, we are creating a new Core Facility, the Smallpox Vaccine Clinical Research Center, to coordinate and facilitate clinical/translational research on smallpox for the MRCE. The Smallpox Vaccine Clinical Research Center will serve two allied missions. First, it will be the clinical/translational research arm of the MRCE, facilitating the research designed to answer critical questions about viral shedding, protective immunity, and susceptibility to poxvirus infections (see the Strategic Projects of Storch, Kazura, and Belshe, section C). In this mode it will directly support and execute translational research protocols on projects of special interest to the MRCE mission. The initial protocols will test the efficacy and mechanism of action of vaccinia immune globulin; an additional project is to fully characterize the immunologic responses of a non-responder to vaccinia immunization. Standard research protocols for characterizing individuals with severe adverse effects to smallpox vaccination will also be supported. The Smallpox Vaccine Clinical Research Center will serve its second mission by becoming the regional center for the diagnosis and care of individuals with serious adverse effects after smallpox vaccination. In addition to providing state of the art diagnostics and care, for these individuals, when appropriate, the Smallpox Vaccine Clinical Research Center will offer to enroll them into clinical/translational protocols on the adverse effects of smallpox vaccination. If possible, these individuals will undergo many of the detailed virologic, immunologic, and genetic studies outlined in the projects of Drs. Storch, Kazura, and Belshe. Some of these individuals may have immunodeficiencies (e.g. those with generalized vaccinia or progressive vaccinia) and these will be fully characterized. In this capacity the Smallpox Vaccine Clinical
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Research Center will also become a database for adverse effects to smallpox vaccine in this area, and facilitate any national efforts in this area. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •
Project Title: THERAPEUTICS FOR POX, FILO AND OTHER VIRAL PATHOGENS Principal Investigator & Institution: Schneller, Stewart W.; Professor; Chemistry; Auburn University at Auburn Auburn University, Al 36849 Timing: Fiscal Year 2003; Project Start 01-AUG-2003; Project End 31-JAN-2008 Summary: (provided by applicant): The possibility for biological terrorism has moved from the realm of speculation into reality. As a consequence, the National Institute of Allergy and Infectious Diseases has identified a number of agents that pose the greatest threat and has recommended that therapeutic agents be developed against them. Among that group of pathogens are smallpox and the viral hemorrhagic fevers (including the filoviruses Ebola and Marburg). In preparation for possible release of smallpox, plans are well underway to avail sufficient quantities of smallpox vaccine for a large number of people. However, within this population, there are individuals who will not be able to tolerate the vaccine or, for pre-existing health and/or age reasons, will not be suitable to receive vaccine protection. Thus, for these and other reasons, therapeutic agent development has been recommended by the NIAID. This proposal deals with two therapeutic goals in this direction: (i) agents for treating smallpox infection for instances where the vaccine was ineffective, not available or not administered for reasons just outlined, and (ii) drugs to counteract the effects of vaccination when administered. For the filoviruses Ebola and Marburg, which manifest themselves differently than smallpox but still produce horrendous effects, there are currently no pre-infection vaccines nor post-infection therapeutics available. To address these situations, plans are put forth for designing and developing drugs that would act by inhibiting viral replicative steps following infection by these viruses. Focus will be on nucleosides and nucleotides that affect, primarily, nucleic acid metabolism. A consortium of 3 chemists and 10 virologists has been assembled for this purpose. Because of the extensive collaborations that have been ongoing for some time by this group of chemists and virologists, it can be expected that promising anti-orthopoxvirus and anti-filovirus agents will be uncovered. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen
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Project Title: VACCINIA VIRUS-SPECIFIC T-CELL PHENOTYPES Principal Investigator & Institution: Rock, Michael T.; Medicine; Vanderbilt University 3319 West End Ave. Nashville, Tn 372036917 Timing: Fiscal Year 2004; Project Start 01-APR-2004; Project End 31-MAR-2006 Summary: (provided by applicant): Vaccinia virus, despite its high level of reactogenicity, remains the only available option for protection against smallpox infection. Due to concern about the potential use of smallpox virus as a bioweapon, smallpox vaccination currently is being reintroduced. However, the immune defense mechanisms protecting a person from smallpox after vaccination are not completely understood. The smallpox vaccine provides a unique opportunity to evaluate T cell responses in truly separate cohorts (naive and experienced) to begin delineating the phenotypes and functional characteristics of these cells in protection from disease. Using state-of-the-art polychromatic flow cytometry, we can now examine a multitude of lymphocyte subsets in the peripheral blood that previously were inaccessible, each with a unique functional and phenotypic profile. By studying patients with adverse events,
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we may be able to assign clinical relevance to some of these subsets. The goal of this research program is 1) to identify the functional and phenotypic profile of vaccinia virus-specific T cells elicited following smallpox immunization and 2) to examine the persistence of vaccinia-specific T cells in vaccinia-experienced vaccinees and the breadth of the immune response in immune and non-immune subjects by determining the Vbeta repertoire usage by those T lymphocytes. Distinct T cell subsets respond differently to viral infections and characterization of these subsets may lead to a better understanding of the immune response and the role of these cells in shaping long-term memory responses. Thus, we will learn not only about immunity to vaccinia, but also learn about the basic biology of virus-specific T cell responses. This work will provide a foundation for comparison of the cellular immunogenicity induced by other candidate vaccines, such as further attenuated viruses like MVA, or cell culture derived vaccinia. Given that a true efficacy trial cannot be conducted with these vaccines, likely correlates of immunity are needed for comparative purposes. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •
Project Title: VIROLOGY AND SMALLPOX VACCINATIONS Principal Investigator & Institution: Storch, Gregory; Washington University Lindell and Skinker Blvd St. Louis, Mo 63130 Timing: Fiscal Year 2003; Project Start 01-AUG-2003; Project End 31-JUL-2008 Summary: The broad objective of this project is to use conventional and molecular techniques to define the virologic events following smallpox vaccination in vaccinianaive and vaccinia-experienced individuals. The specific alms are to: 1) Define the virologic events associated with smallpox vaccination. 2) Determine whether multiple viral variants are present within the Dryvax vaccine, and if so, to investigate their role in the virology of smallpox vaccination and in adverse reactions. 3) Define the virologic events associated with adverse reactions to smallpox vaccination. 4) Examine the virologic response to treatment with vaccinia immune globulin (VIG) and/or cidofovir in vaccinees who require these therapies to control adverse reactions. A quantitative real-time PCR assay will be developed and used to measure the level of vaccinia DNA at regular intervals after vaccination. Specimens will also be cultured for vaccinia virus. These studies will be useful for defining the possible contagiousness of individuals having smallpox vacciniation and for helping determine the need for donor deferral for voluntary blood donations. The data will also provide a basis for studies of the immunology and immunogentics of vaccinia. Studies will be performed of Dryvax vaccine to define variants within the vaccine virus. In collaboration with the Genome Sequencing Center, the complete nucleotide sequence of 5 variant strains will be determined. Specific assays will be developed and used to define the contribution of variants to immunogenicity and reactogenicity of the vaccine. Smallpox adverse reaction clinics will be established at each participating medical center to evaluate individuals with possible adverse reactions. Individuals seen in these clinics will be recruited to participate in detailed studies of the virology, immunology, and immunogenetics of smallpox vaccination. These studies will investigate the virology of adverse reactions, the relationship between viral and immunologic events, and the genetic basis for both. For individuals having severe adverse reactions, virologic studies will be used to help evaluate and guide therapy with VIG and cidofovir. The studies described will form a basis for evaluating Dryvax as well as future smallpox vaccines. The assays to be developed and the clinics to be established will provide an infrastructure that will be available to respond to a bioterrorist attack on the United States. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen
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Project Title: VULNERABILITY TO SMALLPOX DUE TO DECLINING CTL IMMUNITY Principal Investigator & Institution: Mbawuike, Innocent N.; Molecular Virology & Microbiol; Baylor College of Medicine 1 Baylor Plaza Houston, Tx 77030 Timing: Fiscal Year 2002; Project Start 01-SEP-2002; Project End 31-AUG-2004 Summary: (provided by applicant): Discontinuation of smallpox vaccination since 1971 has led to waning of acquired immunity in the U.S. general population, thus, raising the risk of major epidemics from intentional release of smallpox by terrorists or unfriendly governments. The CDC has shown that vaccinia immunization, which led to the eradication of smallpox worldwide, caused excess rates of severe complications and death among older persons. Since higher morbidity and mortality rates from many other viral infections in older persons have been attributed to a deficiency in virus-specific HLA-restricted CTL response, the elderly will be expected to possess reduced vacciniaspecific CTL activity and therefore be more susceptible to smallpox disease as well as complications from vaccinia immunization. The goals of this study are to: 1) profile the sero-prevalence and memory CD8+ CTL activity status against vaccinia in representative young adult and elderly U.S. populations, 2) identify easily detectable and quantifiable surrogates of CTL competence against smallpox infection and 3) explore immunological basis for complications from vaccinia vaccination. Levels of vaccinia-specific memory CD8 CTL activity will be determined in peripheral blood lymphocytes of U.S. born healthy adults (35-49 and 50-64 years old) and elderly adults (~65 years old) who have previously been vaccinated with vaccinia and compared with young adults (18-32 years old) who are recent (
