MULTI-INTRACELLULAR PATHOGEN EPITOPE-BASED VACCINE

Anne Searls De groot, Research Professor
University Of Rhode Island, Research Office, Kingston, Ri 02881-0811

Abstract: In the context of this proposal, we will use pre-existing defined epitopes for Fransicella tularensis (FT), and using the core TRIAD vaccine design toolkit, define new epitopes for Burkholderia pseudomallei (BPM) and Burkholderia mallei (BM), the agents of meloidosis and glanders, respectively, for use in an epitope-based multipathogen prophylactic vaccine. FT has been listed as a Category A biological warfare agent as a result of World War II and Cold War-era biowarfare research. BPM, the etiological agent of melioidosis, is responsible for an estimated 20% of septicemias and approximately 40% of deaths due to bacterial sepsis in tropical regions of the world. BM, a related bacterium, also causes fatal infections (classified as glanders) in man and animals. Like FT, BM is highly infectious as an aerosol. All three pathogens (FT and BPM/BM) are intracellular bacteria and thus amenable to attack by cell-mediated immune response. The EpiMatrix epitope-based vaccine design platform has already yielded a prototype F. tularensis Type A (subsp. tularensis SCHU S4) vaccine that confers 60% protection against heterologous lethal respiratory challenge with the live vaccine strain (LVS), an attenuated subsp. holarctica derivative. To our knowledge no subunit vaccine for tularemia has achieved a comparable level of protection in this well-developed lethal respiratory challenge model. This milestone was reached over the course of a 24 month funding period. The same vaccine design tools, made available in the context of this U19 program project, will facilitate the development a novel combined vaccine against the three pathogens. We will test the combined vaccine components, and optimize dose, delivery vehicle, and adjuvants, in a live challenge model. In addition to evaluating our epitope-driven vaccine, we will explore whether combining our FT/BPM/BM multi-pathogen vaccine with the anti-LPS vaccine developed by Dr. Steven Opal and colleagues will lead to improved protection against live challenge. The challenge studies will be carried out at NERCE in collaboration with Brown University (Steve Gregory, Steve Opal) investigators. This milestone-driven program will lead to proof-of-principle (evidence for protection against live challenge) and development of a licensable multi-pathogen biodefense vaccine within a five year time frame

Budget start date: 1-JUL-2010

Budget end date: 30-JUN-2011

5U19AI082642-02_8114 (2010): $225532

Sponsored Links Excellgen http://Excellgen.com

	Recombinant Lentivirus & Adenovirus High Yield and High Titer up to 10¹⁰ (lentivirus) and 10¹³ (adenovirus) for Guaranteed Expression of GOI. ~~$3000~~, $2500
	Transient Protein Expression in CHO and HEK293 Cells Transient Expression, Truly Functional Protein, 95% purity, 1~20 mg, fast turnaround. ~~$5500~~, $3950
	Baculovirus Protein Expression Fast turn around, >95% purity functional protein. No outsourcing to China or India. ~~$5500~~, $3950

TRANSLATIONAL IMMUNOLOGY RESEARCH AND ACCELERATED [VACCINE] DEVELOPMENT (TRIAD)

Anne Searls De groot, Research Professor
University Of Rhode Island, Research Office, Kingston, Ri 02881-0811

Grant 5U19AI082642-02 from National Institute Of Allergy And Infectious Diseases

Abstract: To meet the dual threats of emerging infectious diseases and engineered biowarfare/bioterror agents, there is a pressing need for more efficient systems for vaccine development. TRIAD, or the Translational Immunology Research and Accelerated [Vaccine] Development program, based in the Biotechnology Program at the University of Rhode Island, has pioneered the development and application of an integrated "gene to vaccine" in silico, in vitro and in vivo vaccine design program to address this need. TRIAD has selected Category A pathogens F. tularensis, Category B agents Burkholderia pseudomallei and Burkholderia mallei, and emerging infectious diseases (HCV, H. pylori, tick borne diseases) as the focal point of this proposal. Using the TRIAD immunoinformatics Toolkit, TRIAD investigators will pursue the development of second generation epitope based immunme-derived vaccines for these pathogens, while addressing the failings of prior generations of epitope based vaccines. We will maximize payload quantity using validated immunoinformatics tools that permit selection of optimal T cell epitopes that are highly conserved and immunogenic. We will ensure payload quality by choosing epitopes that demonstrate antigenicity in human PBMC as well as protection in established murine models of disease/infection. We will select a combination of promiscuous Class II epitopes, and Class I supertype epitopes will provide >99% coverage of human populations. We will avoid cross-reactive epitopes and explore the role of regulatory T cells in the context of improving vaccine design. Where appropriate, we will combine our epitope-driven vaccines with broad-spectrum anti-LPS vaccines. We will optimize payload, delivery, formulation, and adjuvanting by exploring a range of delivery options [Dendritic cells, DEC205, DNA, electroporation, mucosal delivery). The TRIAD project aims to develop vaccines demonstrating broad spectrum activity include crossprotective and multiple component vaccines, and delivery technologies that have the potential to be effective against multiple emerging and re-emerging infectious diseases. Our efforts to merge rational design with recent advances in vaccine delivery will manifest in a coordinated toolkit and a cadre of informed users, who will be ready and able to apply the tools to discover new treatments for emerging infectious disease and biodefense. RELEVANCE The Translational Immunology Research and Accelerated [Vaccine] Development (TRIAD) consortium will join forces to accelerate the development of safer, more efficient vaccines for biodefense and emerging infectious diseases. The TRIAD will also support the development of vaccine design methods that significantly reduce the time and cost required to bring medical countermeasures to market. PROJECT 1 Multi-intracellular Pathogen Epitope-based Vaccine (De Groot, A) PROJECT 1 In the context of this proposal, we will use pre-existing defined epitopes for Francisella tularensis (FT), and using the core TRIAD vaccine design toolkit, define new epitopes for Burkholderia pseudomallei (BPM) and Burkholderia mallei (BM), the agents of meloidosis and glanders, respectively, for use in an epitope-based multipathogen prophylactic vaccine. FT has been listed as a Category A biological warfare agent as a result of World War II and Cold War-era biowarfare research. BPM, the etiological agent of meloidosis, is responsible for an estimated 20% of septicemias and approximately 40% of deaths due to bacterial sepsis in tropical regions of the world. BM, a related bacterium, also causes fatal infections (classified as glanders) in man and animals. Like FT, BM is highly infectious as an aerosol. All three pathogens (FT and BPM/BM) are intracellular bacteria and thus amenable to attack by cell-mediated immune response. The EpiMatrix epitope-based vaccine design platform has already yielded a prototype F. tularensis Type A (subsp. tularensis SCHU S4) vaccine that confers 60% protection against heterologous lethal respiratory challenge with the live vaccine strain (LVS), an attenuated subsp. holarctica derivative. To our knowledge no subunit vaccine for tularemia has achieved a comparable level of protection in this well-developed lethal respiratory challenge model. This milestone was reached over the course of a 24 month funding period. The same vaccine design tools, made available in the context of this U19 program project, will facilitate the development a novel combined vaccine against the three pathogens. We will test the combined vaccine components, and optimize dose, delivery vehicle, and adjuvants, in a live challenge model. In addition to evaluating our epitope-driven vaccine, we will explore whether combining our FT/BPM/BM multi-pathogen vaccine with the anti-LPS vaccine developed by Dr. Steven Opal and colleagues will lead to improved protection against live challenge. The challenge studies will be carried out at NERCE in collaboration with Brown University (Steve Gregory, Steve Opal) investigators. This milestone-driven program will lead to proof-of-principle (evidence for protection against live challenge) and development of a licensable multi-pathogen biodefense vaccine within a five year time frame. RELEVANCE Fransicella tularensis, Burkholderia pseudomallei and Burkholderia mallei are included on the list of Category A (FT) and B bioterrorist agents (BPM, BM) due to concern about their potential for weaponizaton. Development and licensure of effective and safe vaccines for these biowarfare threats remains a valid but unmet NIAID and DoD objective

Keywords: 2nd World War; Address; Adjuvant; Advocate; Aerosols; Animals; Antigenic Determinants; Appointment; Attenuated; Attenuated Vaccines; B. mallei; B. pseudomallei; Bacteria; Binding Determinants; Biologic Sciences; Biologic Warfare; Biological Sciences; Biological Warfare; Biotechnology; Blood Poisoning; Burkholderia mallei; Burkholderia pseudomallei; C. pylori; Campylobacter pylori; Categories; Cells; Center for Translational Science Activities; Cessation of life; Collaborations; Collection; Combined Vaccines; Communicable Diseases, Emerging; Communities; Computer Simulation; Computerized Models; DNA; DNA Molecular Biology; Death; Dendritic Cells; Deoxyribonucleic Acid; Development; Disease model; Doctor of Medicine; Doctor of Philosophy; Dose; Drug Formulations; Educational workshop; Electroporation; Emerging Communicable Diseases; Engineering; Engineerings; Ensure; Environment; Epitopes; Epitopes, T-Lymphocyte; F. tularensis; Formulation; Formulations, Drug; Francisella tularensis; Funding; Generations; Genes; Glanders; Goals; Grant; H. pylori; H. pylory; H.pylori; HCV; HCV infection; Hand; Helicobacter pylori; Hepatitis C; Hepatitis C virus; Hepatitis C virus infection; Hepatitis, Viral, Non-A, Non-B, Parenterally-Transmitted; Hepatitus C; Human; Human, General; Immunology; Immunology (Including BRMP); Immunology (NCI Program); In Vitro; Infection; Infectious Diseases, Emerging; Informatics; Institutes; Internet; Investigators; Knowledge; Lead; Licensure; Life; Life Sciences; M.D.; Mammals, Mice; Man (Taxonomy); Man, Modern; Marketing; Mathematical Model Simulation; Mathematical Models and Simulations; Medical; Methods; Methods and Techniques; Methods, Other; Mice; Modeling; Models, Computer; Molecular Biology; Murine; Mus; NANBH; NIAID; National Institute of Allergy and Infectious Disease; P. mallei; P. pseudomallei; P.mallei; P.pseudomallei; PBMC; PT-NANBH; Parenterally-Transmitted Non-A, Non-B Hepatitis; Pasteurella tularensis; Pb element; Peripheral Blood Mononuclear Cell; Ph.D.; PhD; Population; Programs (PT); Programs [Publication Type]; Pseudomonas mallei; Pseudomonas pseudomallei; Regulatory T-Lymphocyte; Research; Research Personnel; Researchers; Rhode Island; Role; Second World War; Sepsis; Septicemia; Simulation, Computer based; Subunit Vaccines; System; System, LOINC Axis 4; T-Cell Epitopes; T-Lymphocyte Epitopes; Techniques; Technology; Testing; Tick-Borne Diseases; Time; Toxin; Training; Training Programs; Tularemia; Universities; Vaccine Design; Vaccines; Vaccines, Attenuated; Vaccines, Combination; Vaccines, Combined; Veiled Cells; WWW; War; Workshop; World War II; World War, 1939-1945; base; biodefense; biowarfare; bloodstream infection; cell mediated immune response; college; computational modeling; computational models; computational simulation; computer based models; computerized modeling; computerized simulation; conference; cost; design; designing; disorder model; heavy metal Pb; heavy metal lead; hepatitis non A non B; hepatitis nonA nonB; immunogenic; improved; in silico; in vivo; live vaccine; man; man`s; meetings; member; new vaccines; next generation vaccines; non A non B hepatitis; non A, non B hepatitis; non-A non-B hepatitis; non-A, non-B hepatitis; novel; novel vaccines; pathogen; professor; programs; prophylactic; prototype; respiratory; septicaemia; skills; social role; symposium; theories; tool; vaccine delivery; vaccine development; virtual simulation; web; world wide web

Project start date: 2009-07-20

Project end date: 2014-06-30

Budget start date: 1-JUL-2010

Budget end date: 30-JUN-2011

PFA/PA: RFA-AI-08-014

5U19AI082642-02 (2010): $2564398

TRIAD ADMINISTRATIVE CORE

Anne Searls De groot, Research Professor
University Of Rhode Island, Research Office, Kingston, Ri 02881-0811

Budget start date: 1-JUL-2010

Budget end date: 30-JUN-2011

5U19AI082642-02_8119 (2010): $238544

TRIAD PILOT PROJECTS CORE

Anne Searls De groot, Research Professor
University Of Rhode Island, Research Office, Kingston, Ri 02881-0811

Budget start date: 1-JUL-2010

Budget end date: 30-JUN-2011

5U19AI082642-02_8120 (2010): $302239

Grants awarded to Anne Searls De groot

T1D Tolerance Induction With Natural Treg Epitopes

Anne Searls De groot
Epivax, Inc.
providence, Ri 02903

Grant 1R43DK081261-01 from National Institute Of Diabetes And Digestive And Kidney Diseases IRG: ZRG1

Abstract: Type 1 (juvenile) diabetes is an organ-specific autoimmune disease resulting from destruction of insulinproducing pancreatic beta-cells. In non-diabetics, islet cell antigen-specific T cells are either deleted in thymic development or are converted to T regulatory cells that actively suppress effector responses to islet cell antigens. In juvenile diabetics and in the NOD mouse model of juvenile diabetes, these tolerance mechanisms are missing. In their absence, islet cell antigens are presented by human leukocyte antigen (HLA) class I and II molecules and are recognized by CD8(+) and CD4(+) auto-reactive T cells. Destruction of islet cells by these auto-reactive cells eventually leads to glucose intolerance. Modulation of auto-immune responses to autologous epitopes by induction of antigen-specific tolerance may prevent ongoing beta-cell destruction. EpiVax, Inc. has identified a set of natural T regulatory epitopes that induce tolerance to immunogenic proteins. Preliminary studies demonstrate that these "Tregitopes" specifically induce natural Tregs and, when coadministered with an antigen, lead to expansion of antigen-specific regulatory T cells. Initial studies have demonstrated tolerance induction in the context of dust-mite allergy. The goal of this Phase I SBIR application is to evaluate whether immune response to islet cell antigens can be redirected by administration of Tregitopes in conjunction with T1D antigens, leading to "antigen-specific adaptive tolerance induction" (T1D ASATI) in a murine model of T1D. The outcome of T1D ASATI will be measured in terms of regulatory T cell responses to preproinsulin (PPI) T-cell epitopes in vivo and in vitro. In the context of this application we will (1) Assess whether Tregitopes suppress the PPI-specific response ex vivo using PBMC from recent-onset type 1 diabetics, (2) Measure Tregitope-induced modulation of T1D pathology and PPI-specific immune responses in NOD mice and (3) Elucidate the mechanism of Tregitopeinduced PPI ASATI in DR4 transgenic mice. If successful, this research and development program will lead to a clinical approach that will generate or restore tolerance in juvenile diabetics. This research will investigate an immune treatment that may slow the progression of Type 1 (juvenile) diabetes

Project start date: 2008-07-01

Project end date: 2010-06-30

1R43DK081261-01 (2008): $493560

STUDIES OF REACTIVATION TUBERCULOSIS IN HIV INFECTION

Anne Searls De groot
Miriam Hospital
providence, Ri 029062853

Grant 5K08AI001068-03 from National Institute Of Allergy And Infectious Diseases IRG: AIDS

Abstract: Tuberculosis (TB), a major public health problem worldwide, is undergoing a resurgence in the United States, largely as a result of TB reactivation in patients infected with the Human Immunodeficiency Virus-1 (HIV) New information is needed for development of novel TB control strategies. The clinical immunology studies described in this proposal are directed toward the long-range goal of gaining greater understanding of host defenses in TB. At the same time, the results of these studies should provide some new insights into the immunopathogenesis of AIDS. To assess the possibility that TB reactivation is due to the development of qualitative defects in certain immune responses in HIV+ patients, HIV- infected (HIV+) and uninfected (HIV-) subjects who have no TB (PPD-), latent TB (PPD+) or active TB will be compared for a variety of relevant clinical and immunological parameters including quantitative T cell subsets; in vitro lymphoproliferative responses to Mycobacterium tuberculosis (Mtb) and unrelated antigens and lectin mitogens; production of selected cytokines (interferon-gamma, TNF, and other) in response to Mtb antigens; cell-mediated cytotoxicity to cells bearing Mtb antigens; and activation of intracellular cytostasis of Mtb. In addition, selected individuals from the HIV+/PPD+ group will be studied in a longitudinal manner to assess effects of TB reactivation on these parameters. Statistical analysis carried out during the course of these investigations will incorporate appropriate consideration of stratification and "power". Subjects (primarily prisoners, intravenous drug users, Blacks, Hispanics, and women -- populations at particular risk for dual HIV-TB infection) will be recruited at a state hospital. Based on the results of these studies, future experiments are planned to predict the T cell epitopes of sequenced Mtb protein antigens and test subjects´ lymphocyte responses to the corresponding synthetic peptides. These investigations may reveal selected qualitative defects associated with TB reactivation in HIV disease, shedding light on host defenses that maintain TB latency. The findings may eventually contribute to the design of an effective synthetic subunit vaccine against Mtb that could be used in immunocompromised hosts such as subjects with HIV infection

Keywords: HIV infection, Mycobacterium tuberculosis, human immunodeficiency virus 1, relapse /recurrence, tuberculosis bacterial antigen, bacterial protein, cell mediated cytotoxicity, correctional institution, cytotoxic T lymphocyte, epitope mapping, helper T lymphocyte, host organism interaction, human population study, immunologic memory, interferon, interleukin 2, intravenous drug abuse, leukocyte activation /transformation, monocyte, opportunistic infection, protein sequence, tumor necrosis factor alpha African American, Hispanic American, female, fluorescence activated cell sorter, human subject, peptide chemical synthesis, tissue /cell culture

Project start date: 1992-01-01

Project end date: 1993-09-30

5K08AI001068-03 (1993): $77490

REACTIVATION TUBERCULOSIS IN HIV INFECTION

Anne Searls De groot
Miriam Hospital Providence, Ri 029062853

Grant 7K08AI001068-02 from National Institute Of Allergy And Infectious Diseases IRG: AIDS

Abstract: Tuberculosis (TB), a major public health problem worldwide, is undergoing a resurgence in the United States, largely as a result of TB reactivation in patients infected with the Human Immunodeficiency Virus-1 (HIV) New information is needed for development of novel TB control strategies. The clinical immunology studies described in this proposal are directed toward the long-range goal of gaining greater understanding of host defenses in TB. At the same time, the results of these studies should provide some new insights into the immunopathogenesis of AIDS. To assess the possibility that TB reactivation is due to the development of qualitative defects in certain immune responses in HIV+ patients, HIV- infected (HIV+) and uninfected (HIV-) subjects who have no TB (PPD-), latent TB (PPD+) or active TB will be compared for a variety of relevant clinical and immunological parameters including quantitative T cell subsets; in vitro lymphoproliferative responses to Mycobacterium tuberculosis (Mtb) and unrelated antigens and lectin mitogens; production of selected cytokines (interferon-gamma, TNF, and other) in response to Mtb antigens; cell-mediated cytotoxicity to cells bearing Mtb antigens; and activation of intracellular cytostasis of Mtb. In addition, selected individuals from the HIV+/PPD+ group will be studied in a longitudinal manner to assess effects of TB reactivation on these parameters. Statistical analysis carried out during the course of these investigations will incorporate appropriate consideration of stratification and "power". Subjects (primarily prisoners, intravenous drug users, Blacks, Hispanics, and women -- populations at particular risk for dual HIV-TB infection) will be recruited at a state hospital. Based on the results of these studies, future experiments are planned to predict the T cell epitopes of sequenced Mtb protein antigens and test subjects lymphocyte responses to the corresponding synthetic peptides. These investigations may reveal selected qualitative defects associated with TB reactivation in HIV disease, shedding light on host defenses that maintain TB latency. The findings may eventually contribute to the design of an effective synthetic subunit vaccine against Mtb that could be used in immunocompromised hosts such as subjects with HIV infection.

Keywords: HIV infection, Mycobacterium tuberculosis, human immunodeficiency virus 1, relapse /recurrence, tuberculosis, bacterial antigen, bacterial protein, cell mediated cytotoxicity, correctional institution, cytotoxic T lymphocyte, epitope mapping, helper T lymphocyte, human population study, immunologic memory, interferon, interleukin 2, intravenous drug abuse, leukocyte activation /transformation, monocyte, opportunistic infection, protein sequence, tumor necrosis factor alpha, virus host interaction, African American, Hispanic American, female, fluorescence activated cell sorter, human clinical subject, peptide chemical synthesis, tissue /cell culture

Project start date: 1992-01-01

Project end date: 1994-12-31

7K08AI001068-02 (1992): $37156

Epitope Driven HIV Vaccine Development

Anne Searls De groot
Brown University 164 Angell Street Providence, Ri 02912

Grant 3R01AI050528-03S2 from National Institute Of Allergy And Infectious Diseases IRG: ZRG1

Abstract: Tools developed in the TB/HIV Research Laboratory and recently improved at EpiVax have put the development of a novel, epitope-driven vaccine for HIV-1 within reach. During the course of preparatory research we prospectively identified novel HIV-1 CTL epitopes that are well conserved across HIV-1 clades, produced two DNA vaccine constructs containing these epitopes in a string-of-beads formation, and successfully transfected several constructs in target cells. During the course of this research project, we propose to screen additional CTL and Th epitopes, to refine and improve our vaccine constructs, to confirm the expression and recognition of the epitopes contained within our constructs, and to develop a series of HLA-supertype-homogenous plasmids to a state of readiness for a Phase I trial. Our specific aims for this project will be to 1. Finalize and screen a list of carefully selected candidate HIV-1 class I and class II epitopes 2. Develop plasmids containing these epitopes that target antigen-presenting cells 3. Evaluate the expression of the epitopes by target cells and their recognition in vitro (in cell culture) and in vivo (in transgenic mice) 4. Depending on success of Aims 1-3, initiate pre-clinical safety and toxicity studies and file an IND application with the FDA The long term goal of this research project is to develop HLA-supermotif-homogenous plasmids containing highly conserved epitopes that will be combined in a DNA-plasmid HIV vaccine "cocktail." EpiVax, Inc. will define and select the epitopes in collaboration with the TB/HIV Research Lab; the Lab will screen the epitopes, develop plasmid constructs, and perform in vitro assays confirming the expression of the epitopes by cells transfected with the plasmid constructs. Dr. Weiner s laboratory will test these constructs in the transgenic mouse model (transgenic for A2 and DR 0101, initially). The final product of this joint effort will be a set of plasmids to be combined in an epitope-driven, oligonucleotide-based DNA cocktail vaccine for HIV.

Keywords: AIDS vaccine, antigen, human immunodeficiency virus 1, vaccine development, MHC class I antigen, clinical trial phase I, cytotoxic T lymphocyte, dendritic cell, human therapy evaluation, oligonucleotide, plasmid, vector vaccine, enzyme linked immunosorbent assay, genetically modified animal, human subject, injection /infusion, laboratory mouse, patient oriented research, tissue /cell culture, transfection

Project start date: 2002-05-01

Project end date: 2005-11-30

3R01AI050528-03S2 (2005): $206000

3R01AI050528-02S1 (2003): $51763

5R01AI050528-03 (2004): $1095520

TRANSLATIONAL IMMUNOLOGY RESEARCH AND ACCELERATED [VACCINE] DEVELOPMENT (TRIAD)

Anne Searls De groot, Research Professor
University Of Rhode Island, Research Office, Kingston, Ri 02881-0811

Grant 3U19AI082642-02S1 from National Institute Of Allergy And Infectious Diseases

Project start date: 2010-08-01

Project end date: 2011-07-31

Budget start date: 10-AUG-2010

Budget end date: 31-JUL-2011

PFA/PA: RFA-AI-08-014

3U19AI082642-02S1 (2010): $511121

3U19AI082642-02S2 (2010): $35894

1U19AI082642-01 (2009): $2486017

Sponsored Links Excellgen http://Excellgen.com

	Transient Protein Expression in CHO and HEK293 Cells Transient Expression, Truly Functional Protein, 95% purity, 1~20 mg, fast turnaround. ~~$5500~~, $3950
	Baculovirus Protein Expression Fast turn around, >95% purity functional protein. No outsourcing to China or India. ~~$5500~~, $3950
	Recombinant Lentivirus & Adenovirus High Yield and High Titer up to 10¹⁰ (lentivirus) and 10¹³ (adenovirus) for Guaranteed Expression of GOI. ~~$3000~~, $2500

TRIAD ADMINISTRATIVE CORE

Anne Searls De groot, Research Professor
University Of Rhode Island, Research Office, Kingston, Ri 02881-0811

Keywords: Address; Administrator; Advocate; Appointment; B. mallei; B. pseudomallei; Bears; Biologic Sciences; Biologic Warfare; Biological Sciences; Biological Warfare; Biotechnology; Budgets; Burkholderia mallei; Burkholderia pseudomallei; C. pylori; Campylobacter pylori; Categories; Center for Translational Science Activities; Collection; Commit; Communicable Diseases, Emerging; Communication; Communities; Computer Simulation; Computerized Models; Data; Development; Educational workshop; Emerging Communicable Diseases; Engineering; Engineerings; Ensure; Environment; F. tularensis; Fostering; Francisella tularensis; Generations; Genes; Goals; Grant; Group Meetings; H. pylori; H. pylory; H.pylori; HCV; HCV infection; Hand; Health; Helicobacter pylori; Hepatitis C; Hepatitis C virus; Hepatitis C virus infection; Hepatitis, Viral, Non-A, Non-B, Parenterally-Transmitted; Hepatitus C; Human; Human, General; Immunology; Immunology (Including BRMP); Immunology (NCI Program); In Vitro; Infectious Diseases, Emerging; Informatics; Institutes; Institution; Investigators; Leadership; Life Sciences; Man (Taxonomy); Man, Modern; Mathematical Model Simulation; Mathematical Models and Simulations; Medical; Meetings, Group; Methods and Techniques; Methods, Other; Microbiology; Mission; Models, Computer; NANBH; Outcome; P. mallei; P. pseudomallei; P.mallei; P.pseudomallei; PT-NANBH; Parenterally-Transmitted Non-A, Non-B Hepatitis; Pasteurella tularensis; Programs (PT); Programs [Publication Type]; Pseudomonas mallei; Pseudomonas pseudomallei; Publications; Reagent; Research; Research Personnel; Research Resources; Researchers; Resource Sharing; Resources; Rhode Island; Science of Microbiology; Scientific Publication; Simulation, Computer based; System; System, LOINC Axis 4; Techniques; Technology; Teleconferences; Tick-Borne Diseases; Time; Toxin; Training; Training Programs; Transmission; Travel; Triad; Triad Acrylic Resin; Triad resin; Tularemia; Universities; Ursidae; Ursidae Family; Vaccine Design; Vaccines; Workshop; base; biodefense; biowarfare; college; computational modeling; computational models; computational simulation; computer based models; computerized modeling; computerized simulation; conference; cost; design; designing; hepatitis non A non B; hepatitis nonA nonB; improved; in silico; in vivo; meetings; member; new vaccines; next generation vaccines; non A non B hepatitis; non A, non B hepatitis; non-A non-B hepatitis; non-A, non-B hepatitis; novel vaccines; pathogen; product development; professor; programs; skills; symposium; theories; tool; transmission process; vaccine development; virtual simulation

Relevance: The Administrative core will enable TRIAD to accelerate the development of vaccines for emerging and biowarfare pathogens and educate new generations of vaccine researchers, leading to significant improvements in global health and a radical transformation in the approach to vaccine design around the world

Project start date: 2009-07-20

Project end date: 2014-06-30

Budget start date: 20-JUL-2009

Budget end date: 30-JUN-2010

PFA/PA: RFA-AI-08-014

1U19AI082642-01_8119 (2009): $197051

Optimization Of A Multivalent Tuberculosis Vaccine

Anne Searls De groot
Epivax, Inc.
providence, Ri 02903

Grant 1R43AI075830-01 from National Institute Of Allergy And Infectious Diseases IRG: ZRG1

Abstract: This new Phase I SBIR proposal addresses the continuing worldwide need for a tuberculosis (TB) vaccine. We detail a novel multivalent strategy that aims to elicit immunity to prevent reactivation of latent TB. This epitope-driven, DNA-prime, protein-boost TB vaccine has been in development since 1997, when our immunoinformatics tools were first applied to identify T cell epitopes from TB proteins. In progress made during the three years of NIH and Sequella/Aeras TB Foundation support, we completed mapping of three sets of TB epitopes including novel epitopes predicted directly from the TB CDC1551 genome. In the next phase of the research program, building on our own experiences and our collaborations, we seek to develop the optimal vaccination strategy, using HLA transgenic mice as the model for in vivo study. Before the start of the performance period, we will make the final epitope selections. In the course of the SBIR award period, selected epitopes will be formulated as DNA and peptide/protein vaccines. By means of a prime-boost vaccination strategy, we will optimize key vaccination parameters (administration route, antigen targeting, adjuvant) to induce the greatest immunogenicity as assessed by cytokine production of stimulated immune cells. Next, we will determine the protective efficacy of the optimized TB vaccine against the standard bacilli Calmette-Gurin (BCG) vaccine and as a boost in HLA transgenic mice pre-vaccinated with BCG. Successful completion of this work will set the stage for Phase II development that will partner the vaccine with improved recombinant BCG vaccines and assess efficacy in additional strains of HLA transgenic mice

Project start date: 2007-09-01

Project end date: 2009-08-31

1R43AI075830-01 (2007): $299908

NATURAL AND PREDICTED ANTIGENS FOR TB IMMUNOTHERAPY

Anne Searls De groot
Brown University 164 Angell Street Providence, Ri 02912

Grant 5R01AI035271-03 from National Institute Of Allergy And Infectious Diseases IRG: SRC

Abstract: There has never been a greater need for research on new preventative and therapeutic vaccines to prep and treat tuberculosis (TB). BCG, the only currently available vaccine, has variable efficacy and may not be safe in the context of HIV infection. The morality rate from TB is increasing as multi-drug resistant strains of Mycobacterium tuberculosis (Mtb) become more prevalent. The co-prevalence of HIV and TB in the United States and in developing countries will contribute to the acceleration of the TB case rate worldwide. Advances in TB prevention and immunotherapy must be based on identification of the immunodominant epitopes of antigens that promote "protective" immune responses. In this application, I outline a proposal to identify Mtb antigens that are associated with a protective pattern of response in Mtb-infected patients, using two novel experimental approaches. I propose to identify Mtb protein epitopes by extracting "natural peptides" from antigen presenting cells infected with live, virulent Mtb. Additional Mtb epitopes will be identified by screening published Mtb protein sequences for amino acid sequences that conform to established "natural" motifs and/or to a computerized algorithin (AMPHI). These putative epitopes will be-synthesized and tested for immunogenicity in in vitro assays using cells and cell lines derived from Mtb-immune subjects. Peptide epitopes that appear to be immunogenic will then be evaluated in assays designed to compare the immune responses of Mtb-immune and Mtb- susceptible subjects. Peptides that provoke T cell proliferation or CTL responses in cells derived from Mtb-immune subjects, but not in cells derived from Mtb-susceptible subjects, will be considered "protective" antigens, candidates for inclusion in a synthetic multi-subunit TB vaccine. I will also explore the potential use of "protective" antigens in the development of an adoptive immunotherapy technique for the treatment of multi-drug resistant TB. Preliminary investigations described in this application demonstrate a decrease of Mtb-specific T cell proliferation responses to several mycobacterial protein preparations (PPD, lyophilized Mtb) in Mtb- susceptible subjects. I also present results from comparisons of Mtb- specific cytotoxicity in cell lines derived from Mtb-immune and Mtb- susceptible individuals. The isolation of natural peptide epitopes and the identification of peptides by algorithm will be performed in collaboration with Dr. Jay A. Berzofsky of the National Cancer Institute, NIH. Dr. Judy Lieberman of New England Medical Center will collaborate on the development of cell lines for adoptive immunotherapy. The in vitro assays will be performed in the Biohazard level 3 facility at Brown University. The study population will include patients with latent Mtb infection and active TB disease, who will be recruited in three public hospital settings. Indigent individuals, Blacks, and Hispanics will make up a large proportion of the study population.

Project start date: 1993-09-30

Project end date: 1997-06-30

5R01AI035271-03 (1995): $245074

5R01AI035271-02 (1994): $215579

1R01AI035271-01 (1993): $215179

OPTIMIZATION OF HIV VACCINE SUBUNIT DELIVERY

Anne Searls De groot
Epivax, Inc., Providence, Ri 02903

Grant 3R21AI078800-02S2 from National Institute Of Allergy And Infectious Diseases

Abstract: The goal of this new R21 proposal is to develop a pro-inflammatory and non-tolerogenic HIV vaccine delivery system based on the dendritic cell targeting anti-DEC-205 antibody. The success of anti-DEC-205 as a stimulator of strong inflammatory immune responses depends on co-administration of non-specific dendritic cell maturation factors. In their absence, anti-DEC-205 induces antigen-specific tolerance rather than immunity. We hypothesize that regulatory T-cell epitopes contained in anti-DEC-205 promote a tolerogenic reaction that is only overcome through the co-administration of non-specific immuno-stimulators. This hypothesis is based on our discovery of a set of natural regulatory T-cell epitopes derived from human immunoglobulins that induce tolerance by stimulating regulatory T cells. We have verified experimentally that these epitopes cause antigen-specific expansion of regulatory T cells and suppress inflammatory immune responses. We propose to develop a modified pro-inflammatory and non-tolerogenic anti-DEC-205 antibody. We expect that modification of regulatory T-cell epitopes will significantly diminish tolerogenicity, enabling use of anti-DEC-205 as a stand-alone HIV antigen delivery system that obviates the dangers associated with non-specific activation of the immune system. We will de-tolerize anti-DEC-205 by epitope modification using the process we developed to reduce immunogenicity of protein therapeutics. We will substitute key amino acids in the regulatory T-cell epitopes with those that are experimentally shown to interfere with MHC binding. We will then (1) produce a set of antibody variants recombinantly conjugated to HIV Gag, (2) identify de-tolerizing mutations that do not interfere with dendritic-cell targeting, and (3) and evaluate variants for reduced tolerogenicity, as well as for enhanced Gag immunogenicity. Finally, we will produce and characterize the immunogenicity of a de-tolerized anti-DEC-205-based HIV vaccine composed of immunogenic consensus sequences. This project will develop an improved delivery vehicle for HIV vaccine components. Immune-dampening portions of the delivery vehicle will be silenced so that potent and effective immune responses can be raised against HIV

Keywords: AIDS Antigens; AIDS Virus; APO2; ATGN; Acquired Immune Deficiency Syndrome Virus; Acquired Immunodeficiency Syndrome Virus; Algorithms; Alleles; Allelomorphs; Amino Acids; Antibodies; Antigen Presentation Pathway; Antigen Processing and Presentation; Antigen Targeting; Antigenic Determinants; Antigens; Assay; Binding; Binding (Molecular Function); Binding Determinants; Bioassay; Biologic Assays; Biological Assay; Bone Marrow; CD107a antigen; Cell Growth in Number; Cell Maturation; Cell Multiplication; Cell Proliferation; Cells; Cellular Proliferation; Class II Antigens; Class II Major Histocompatibility Antigens; Confocal Microscopy; Consensus Sequence; ConsensusSequence; Coupled; Cytofluorometry, Flow; DR4; Data; Dendritic Cells; Doctor of Philosophy; Dose; Epitopes; Epitopes, T-Lymphocyte; Exhibits; Exploratory/Developmental Grant; Flow Cytofluorometries; Flow Cytometry; Flow Microfluorimetry; Gagging; Gamma Globulin, 7S; Genetic Alteration; Genetic Change; Genetic defect; Germany; Goals; HIV; HIV Antigens; HIV vaccine; HIV-Associated Antigens; HIV/AIDS Vaccines; HTLV-III; HTLV-III Antigens; HTLV-III-LAV Antigens; Histocompatibility Antigens Class II; Histocompatibility Complex; Histocompatibility Complices; Human; Human Immunodeficiency Viruses; Human T-Cell Leukemia Virus Type III; Human T-Cell Lymphotropic Virus Type III; Human T-Lymphotropic Virus Type III; Human, General; I-A Antigen; Ia Antigens; Ia-Like Antigens; IgG; Immune; Immune Globulins; Immune Response Antigens; Immune response; Immune system; Immune-Response-Associated Antigens; Immunity; Immunoglobulin G; Immunoglobulins; Immunoglobulins / Antibodies; Immunologist; In Vitro; Incubated; Inflammatory; Investigators; LAMP-1; LAV Antigens; LAV-HTLV-III; Label; Lead; Lymphadenopathy-Associated Antigens; Lymphadenopathy-Associated Virus; MGC9365; MHC Class II Molecule; MHC Class II Protein; MHC class II antigen; Major Histocompatibility Complex; Major Histocompatibility Complex Class II; Major Histocompatibility Complices; Mammals, Mice; Man (Taxonomy); Man, Modern; Manuscripts; Measures; Medical center; Mice; Microfluorometry, Flow; Modification; Molecular Interaction; Molecular and Cellular Biology; Murine; Mus; Mutation; Ovalbumin; Pb element; Peptides; Performance; Ph.D.; PhD; Phenotype; Process; Programs (PT); Programs [Publication Type]; Property; Property, LOINC Axis 2; Publications; R21 Mechanism; R21 Program; Reaction; Reflex, Pharyngeal; Regulatory T-Lymphocyte; Research; Research Activity; Research Personnel; Researchers; Reticuloendothelial System, Bone Marrow; Scientific Publication; Staining method; Stainings; Stains; Surface; System; System, LOINC Axis 4; T-Cell Activation; T-Cell Epitopes; T-Cells; T-Lymphocyte; T-Lymphocyte Epitopes; T-Lymphotropic Virus Type III Antigens, Human; TNFRSF10A; TNFRSF10A gene; TRAILR-1; TRAILR1; Testing; Thymus-Dependent Lymphocytes; Transgenic Mice; Universities; Vaccine Design; Vaccine Research; Vaccines; Variant; Variation; Veiled Cells; Virus-HIV; aminoacid; base; body system, allergic/immunologic; design; designing; experience; experiment; experimental research; experimental study; flow cytophotometry; fluorophore; genome mutation; heavy metal Pb; heavy metal lead; host response; human immunodeficiency virus vaccine; immunogen; immunogenic; immunogenicity; immunoresponse; improved; in vitro activity; in vivo; mutant; novel; organ system, allergic/immunologic; prevent; preventing; programs; public health relevance; research study; response; success; therapeutic protein; thymus derived lymphocyte; vaccine delivery

Project start date: 2008-09-30

Project end date: 2011-08-31

Budget start date: 27-SEP-2009

Budget end date: 31-AUG-2011

PFA/PA: PA-06-181

3R21AI078800-02S2 (2010): $243561

3R21AI078800-02S1 (2009): $215016

1R21AI078800-01 (2008): $179000

HIV EVOLUTION AND CELLULAR IMMUNE RESPONSE

Anne Searls De groot
Brown University
164 Angell Street
providence, Ri 02912

Grant 5R01AI040888-04 from National Institute Of Allergy And Infectious Diseases IRG: ARRB

Abstract: Individuals who develop rapid progression of HIV-1 infection have been shown to have a high burden of relatively homogeneous viruses (quasi species) and a low frequency of cytotoxic T lymphocytes that respond to HIV-1, whereas individuals who progress more slowly or not at all have a lower burden of a much more diverse viral quasi species and evidence of vigorous CTL response to HIV-1. We hypothesize that these individuals´ antiviral cellular immune responses (CTL and T helper) are effectively exerting selective pressure upon the viral quasi species, leading to amino acid changes, quasi species diversification, and modified interaction with the host immune system due to altered binding of MHC-binding regions. This proposal describes the development of EpiMatrix, a computer-driven T cell epitope prediction algorithm, and the application of the algorithm to the evaluation of quasi species evolution. It is proposed that the replacement of anchor- based motifs with extended matrix-based motifs (as in EpiMatrix) will improve the predictive capacity of T cell epitope prediction algorithms for selected MHC alleles and will permit evaluation of quasi species evolution. The specific aims of the proposed work are To develop EpiMatrix, a second generation T cell epitope prediction program. To apply EpiMatrix to HIV-1 quasi species; predict, synthesize, & test MHC-binding regions/T epitopes. To evaluate the impact of quasi species evolution on T cell response to epitopes defined in the previous aim. The use of EpiMatrix of predict MHC-binding regions and T cell epitopes within variable regions will allow us, for the first time, to accurately assess the influence of sequence diversity within both conserved and variable regions upon the ability of the host cellular immune response to recognize and respond to HIV-1 quasi species. The successful completion of this project will lead to an enhanced understanding of the interplay between the antiviral pressure of the cellular immune responses, viral burden, and diversity of quasi species within infected individuals. These fundamental interactions that define HIV-1 infection are critical to our understanding of HIV-1 pathogenesis. In addition, the epitope prediction method and epitopes defined during these investigations will aid in the development of a vaccine against AIDS

Keywords: cellular immunity, computer program /software, evolution, human immunodeficiency virus 1, technology /technique development, virus antigen MHC class I antigen, MHC class II antigen, antigen presentation, cytotoxic T lymphocyte, helper T lymphocyte human tissue, peptide chemical synthesis, tissue /cell culture

Project start date: 1997-07-15

Project end date: 2002-06-30

5R01AI040888-04 (2000): $294632

5R01AI040888-03 (1999): $286051

Sponsored Links Excellgen http://Excellgen.com

	Transient Protein Expression in CHO and HEK293 Cells Transient Expression, Truly Functional Protein, 95% purity, 1~20 mg, fast turnaround. ~~$5500~~, $3950
	Baculovirus Protein Expression Fast turn around, >95% purity functional protein. No outsourcing to China or India. ~~$5500~~, $3950
	Recombinant Lentivirus & Adenovirus High Yield and High Titer up to 10¹⁰ (lentivirus) and 10¹³ (adenovirus) for Guaranteed Expression of GOI. ~~$3000~~, $2500

1R01AI040888-01A1 (1997): $304592

MULTI-INTRACELLULAR PATHOGEN EPITOPE-BASED VACCINE

Anne Searls De groot, Research Professor
University Of Rhode Island, Research Office, Kingston, Ri 02881-0811

Keywords: 2nd World War; APO2; Adjuvant; Aerosols; Animal Model; Animal Models and Related Studies; Animals; Antibodies; Antigenic Determinants; Assay; Attenuated; Attenuated Vaccines; B. mallei; B. pseudomallei; Bacteria; Benchmarking; Best Practice Analysis; Binding; Binding (Molecular Function); Binding Determinants; Bio-Informatics; Bioassay; Bioinformatics; Biologic Assays; Biologic Warfare; Biological Assay; Biological Warfare; Blood Poisoning; Burkholderia; Burkholderia mallei; Burkholderia pseudomallei; CCSG; CD8 Cell; CD8 lymphocyte; CD8+ T-Lymphocyte; CD8-Positive Lymphocytes; CD8-Positive T-Lymphocytes; Cancer Center Support Grant; Categories; Cessation of life; Clinical Trials, Phase I; Collaborations; Combined Vaccines; Consult; Core Grant; Cytofluorometry, Flow; DNA; DR1; DR1 gene; DR4; Data Banks; Data Bases; Databank, Electronic; Databanks; Database, Electronic; Databases; Death; Deoxyribonucleic Acid; Development; Differential Display; Displays, mRNA Differential; Dose; Drug Formulations; ELISA; Early-Stage Clinical Trials; Enzyme-Linked Immunosorbent Assay; Epitopes; F. tularensis; Flow Cytofluorometries; Flow Cytometry; Flow Microfluorimetry; Formulation; Formulations, Drug; Francisella tularensis; Funding; Gene Expression; Genes, Class II; Genes, HLA Class II; Genes, MHC Class II; Glanders; Housing; Human; Human, General; Immune response; Immunologist; Immunology; Immunology (Including BRMP); Immunology (NCI Program); In Vitro; Infection; Investigators; Knowledge; Lead; Length of Life; Licensure; Life; Liposomal; Liposomes; Longevity; MGC9365; MHC Class II; MHC Class II Genes; Mammals, Mice; Man (Taxonomy); Man, Modern; Measures; Melioidosis; Mice; Mice, Transgenic; Microfluorometry, Flow; Modeling; Molecular Interaction; Murine; Mus; NC2-Beta; NIAID; National Institute of Allergy and Infectious Disease; Outcome; P. mallei; P. pseudomallei; P.mallei; P.pseudomallei; P30 Grant; PBMC; Pasteurella tularensis; Pb element; Peptides; Peripheral Blood Mononuclear Cell; Phase 1 Clinical Trials; Phase I Clinical Trials; Phase I Study; Phenotype; Population; Production; Programs (PT); Programs [Publication Type]; Proteins; Pseudomonas mallei; Pseudomonas pseudomallei; Relative; Relative (related person); Research; Research Activity; Research Personnel; Researchers; Route; Second World War; Sepsis; Septicemia; Series; Subunit Vaccines; Survivors; T-Cells; T-Lymphocyte; T8 Cells; T8 Lymphocytes; TNFRSF10A; TNFRSF10A gene; TRAILR-1; TRAILR1; Technology; Testing; Thymus-Dependent Lymphocytes; Time; Transgenic Mice; Transgenic Organisms; Triad; Triad Acrylic Resin; Triad resin; Tularemia; Universities; Vaccine Design; Vaccines; Vaccines, Attenuated; Vaccines, Combination; Vaccines, Combined; Vaccines, Subunit; War; World War II; World War, 1939-1945; base; biodefense; biowarfare; bloodstream infection; cell mediated immune response; clinical data repository; clinical data warehouse; cytokine; data repository; design; designing; flow cytophotometry; gene product; genome database; genome sequencing; heavy metal Pb; heavy metal lead; host response; immunogenic; immunogenicity; immunoresponse; improved; in vivo; life span; lifespan; live vaccine; mRNA Differential Displays; man; man`s; model organism; mouse model; novel; pathogen; phase 1 study; phase 1 trial; phase I trial; pre-clinical; preclinical; programs; prophylactic; protective efficacy; protocol, phase I; prototype; relational database; resistant strain; respiratory; septicaemia; success; thymus derived lymphocyte; tool; transgenic; vaccine candidate; vaccine development; vaccine efficacy; vaccine evaluation; vaccine screening; vaccine testing; vector

Relevance: Fransicella tularensis, Burkholderia pseudomallei and Burkholderia mallei are included on the list of Category A (FT) and B bioterrorist agents (BPM, BM) due to concern about their potential for weaponizaton. Development and licensure of effective and safe vaccines for these biowarfare threats remains a valid but unmet NIAID and DoD objective

Project start date: 2009-07-20

Project end date: 2014-06-30

Budget start date: 20-JUL-2009

Budget end date: 30-JUN-2010

PFA/PA: RFA-AI-08-014

1U19AI082642-01_8114 (2009): $199876

EPITOPE-DRIVEN DEIMMUNIZATION OF FACTOR VIII

Anne Searls De groot
Epivax, Inc., Providence, Ri 02903

Grant 3R43HL088834-02S1 from National Heart, Lung, And Blood Institute

Abstract: Hemophilia A patients are prone to develop inhibitory immune responses to the very therapy they require Factor VIII protein replacement. Up to 30% of all hemophiliacs and greater than 50% of severe hemophiliacs produce antibodies (inhibitors) in response to treatment. Immunogenicity to Factor VIII (FVIII) is its most significant complication immunogenicity not only reduces or eliminates the therapeutic efficacy, but also requires that FVIII be delivered by invasive routes since less invasive routes are known to further increase the immunogenicity of therapeutic proteins. To address this problem, it is necessary to develop a less immunogenic FVIII that will provide hemophiliacs with the benefits of prophylactic Factor VIII therapy without the interference of neutralizing antibodies. Neutralizing antibody formation, the root cause of FVIII therapy failure, is a process dependent on antigen presenting cell signaling to helper T cells. We thus propose to develop a de- immunized version of FVIII by T-cell epitope modification. FVIII contains highly immunogenic T- cell epitopes that are excellent targets for mutation in order to prevent T cell-dependent antibody formation. The aims of this Phase I SBIR are to (1) identify and select the key immunodominant T-cell epitopes in FVIII that are responsible for its immunogenicity, (2) strategically modify those epitopes using point amino acid changes as guided by EpiVax´ validated immunoinformatics tools and techniques and (3) to demonstrate that reengineered individual FVIII domains have reduced immunogenicity in a mouse model of hemophilia. A series of FVIII domain variants bearing single epitope modifications will be produced and their immunogenicity will be evaluated. The focus of a Phase II SBIR will be studies of FVIII constructs bearing multiple epitope modifications (combinations of the modifications identified during Phase I). These research and development programs will lead to the development of a commercially available and fully functional Factor VIII protein that has significantly reduced immunogenicity as confirmed in HLA transgenic mice and Factor VIII deficient mice. Epitope-driven deimmunization of factor VIII 12,000 Americans have the blood clotting disorder Hemophilia A, and as such are susceptible to a host of complications from chronic bleeding of the joints to life threatening blood loss due to traumatic injury. This proposal describes a plan to produce an improved clotting protein (Factor VIII) that could be used to treat hemophiliacs since the current treatment is often rejected by the body. The proposed reengineered protein will be evaluated in a mice that have a hemophilia as well as in mice that have a human- like immune system

Project start date: 2007-07-01

Project end date: 2011-06-30

Budget start date: 10-SEP-2010

Budget end date: 30-JUN-2011

PFA/PA: PA-06-134

3R43HL088834-02S1 (2010): $173764

1R43HL088834-01 (2007): $263867

2nd Annual Vaccine Renaissance Conference, RI

Anne Searls De groot
Epivax, Inc. Providence, Ri 02903

Grant 1R13AI069828-01A1 from National Institute Of Allergy And Infectious Diseases IRG: ZAI1

Abstract: Given the global spread of infectious disease, the emergence of virulent new pathogens such as SARS and Avian Flu and the escalating threat of bio-terrorism, there is no better time than now to initiate a Vaccine Renaissance. To that end, The BioGroup of the Rhode Island Tech Collective and EpiVax Collaborative for Vaccine Research and Development (CVRD) will host a two-day conference for leading vaccine researchers and vaccine producers from across the country. This event will take place on June 7, 8, 9 2006 in Providence, Rhode Island. The primary objective of the Vaccine Renaissance 2006 is to provide a forum for the review of current progress in the development of vaccines, including new tools and techniques for accelerating vaccine development. Recent advances in the fields of biodefense vaccines, vaccines against vaccines will be discussed. A second goal of the meeting is to provide a networking opportunity for the vaccine R and D community nationwide, creating important opportunities for collaborations between academic researchers and vaccine industry leaders. National and regional vaccine industry leaders have been invited to speak about recent vaccine developments in their area of expertise. Recent advances by biotech and academic vaccine developers across the country will be presented both as poster and oral presentations. This 2nd Annual conference follows a highly successful 1st Annual Vaccine Renaissance, RI that took place on June 8, 9 2005. More than 80 participants attended that 1st conference from the New England region and beyond.

Keywords: meeting /conference /symposium, vaccine, travel

Project start date: 2006-06-01

Project end date: 2007-05-31

1R13AI069828-01A1 (2006): $17500

Novel Smallpox Vaccine Derived From VV/VAR Immunome

Anne Searls De groot
Epivax, Inc. Providence, Ri 02903

Grant 1R43AI058376-01A1 from National Institute Of Allergy And Infectious Diseases IRG: ZRG1

Abstract: EpiVax specializes in the development of epitope-driven vaccines by screening whole genomes for candidate vaccine components. In this application, we propose to develop a safe, new smallpox vaccine based on epitopes conserved between the vaccinia virus (VV) and Variola (VAR) immunomes that could be used as both a prophylactic and a therapeutic intervention in the event of a bioterrorist attack. First, peptide sequences that are conserved between the vaccinia virus (W) and Variola (Var) genomes will be screened for immunodominant epitopes by computer-driven (EpiMatrix) analysis and confirmed using T cells from VV-immunized individuals. Second, selected epitopes will then be aligned and cloned into DNA vaccine vectors optimized for strong and sustained expression in vivo. Third, we will vaccinate HLA-transgenic mice with the DNA vaccine constructs, measuring de novo immune responses to the DNA vaccine constructs using intracellular cytokine flow cytometry. EpiVax will be responsible for selecting the VV/Var epitopes and managing the project. The TB/HIV Research Lab at Brown University (subcontractor) will perform binding assays and develop the oligonucleotide multi-epitope constructs consisting of strings of the selected epitopes. Dr. Crowe s laboratory (subcontractor, Vanderbilt University) will perform ELISpot assays. Dr. Weiner (subcontractor, Stellar Chance Laboratories, University of Pennsylvania) will perform the vaccination studies. The final product of Phase I will be an evaluation of the immunogenicity and protective effect of several prototype smallpox vaccine constructs in HLA transgenic mice. In Phase II we would screen additional epitopes, build the final vaccine constructs, optimize the immunogenicity of the final vaccine constructs with adjuvants and delivery vehicles, and address the safety, toxicity and immunogenicity of the constructs in human subjects.

Keywords: nonhuman therapy evaluation, smallpox vaccine, vaccine development, vaccine evaluation, vector vaccine, cytotoxic T lymphocyte, drug adverse effect, histocompatibility antigen, immune response, smallpox virus, vaccinia virus, biotechnology, bioterrorism /chemical warfare, clinical research, computer data analysis, enzyme linked immunosorbent assay, green fluorescent protein, human tissue, laboratory mouse, polymerase chain reaction

Project start date: 2005-06-01

Project end date: 2007-05-31

1R43AI058376-01A1 (2005): $499885

5R43AI058376-02 (2006): $527522

A GENOME-DERIVED, EPITOPE-DRIVEN TULAREMIA VACCINE

Anne Searls De groot
Epivax, Inc. Providence, Ri 02903

Grant 5R43AI058326-02 from National Institute Of Allergy And Infectious Diseases IRG: ZRG1

Abstract: EpIVax specializes in the development of epitope-driven vaccines by screening whole genomes for candidate vaccine components. In this application, we describe a rapid approach to the development of a tularemia vaccine that could be for BioDefense. First, F. tularensis genes that encode transmembrane, heat shock and secreted proteins will be screened for immunodominant epitopes by computer-driven (EpiMatrix) analysis of the bacterial genome. Limited in vitro assays will be performed to confirm MHC binding and immunogenicity of representative epitopes. Second, selected epitopes will be aligned and reverse translated in a string-of-beads formation into a DNA construct. The string of beads epitope construct will then be cloned into DNA plasmids optimized for strong and sustained expression in vivo. Third, we will vaccinate HLA-transgenic mice with the DNA vaccine constructs. De novo immune responses to the DNA vaccine constructs will be evaluated using intracellular cytokine flow cytometry. Fourth, we develop a model of protective immunity to aerosolized F. tularensis. Initially we will use this model to benchmark the protective efficacy of the LVS vaccine for later comparison studies. EpiVax will be responsible for selecting the F. tularensis epitopes and managing the project. The TB/HIV Research Lab at Brown University (subcontractor) will develop the oligonucleotide multi-epitope DNA vaccine constructs consisting of strings of the selected epitopes and Dr. Stephen Gregory (subcontractor, Liver Research Center, Lifespan Hospital) will perform the vaccination studies. The final product of Phase I will be an evaluation of the immunogenicity of several prototype tularemia vaccine constructs in transgenic mice and a functional challenge model. In Phase II we would screen additional epitopes, perform F. tularensis challenge studies, build the final vaccine constructs, optimize the immunogenicity of the final vaccine constructs with adjuvants and delivery vehicles and address the safety, toxicity and immunogenicity of the constructs in human subjects.

Keywords: bacterial vaccine, tularemia, vaccine development, vaccine evaluation, vector vaccine, Francisella tularensis, bacterial protein, biotechnology, bioterrorism /chemical warfare, cell mediated lymphocytolysis test, clinical research, enzyme linked immunosorbent assay, flow cytometry, genetically modified animal, human genetic material tag, human subject, laboratory mouse, tissue /cell culture

Project start date: 2004-09-15

Project end date: 2007-08-31

5R43AI058326-02 (2005): $410485

1R43AI058326-01A1 (2004): $421437

A Genome-Derived, Epitope-Driven H. Pylori Vaccine

Anne Searls De groot
Epivax, Inc. Providence, Ri 02903

Grant 5R43AI065036-02 from National Institute Of Allergy And Infectious Diseases IRG: ZRG1

Abstract: EpiVax discovers and develops epitqpe-driven vaccines by screening microbial genomes for protective epitopes. In this application, we describe a genome-based approach for the development of a therapeutic vaccine against Helicobacter pylori disease. First, the H. pylori genome will be scanned for immunodominant Th epitopes by computer-driven (EpiMatrix) analysis. Candidate Th epitopes will be selected based on class II MHC allele restriction and clustering (promiscuity). Epitopes derived from antigens previously shown in vivo as protective (e.g., enzymes, porins, adhesins, heat shock proteins), as well as novel epitopes from proteins not previously implicated in protection will be further sorted on the basis of strain conservation (> 95%), in vivo expression in human and murine gastric tissue, and display of minimal (<30%) sequence homology with human proteins. Peripheral blood leukocytes from H. pylori-infected subjects, and spleen cells from H. pylori-immunized HLA transgenic mice will be used to examine and confirm the ability of the epitopes to activate Th1 and/or Th2 immune responses in vitro. Selected epitopes will be aligned and reverse translated in a string-of-beads array into a DNA construct. The epitope construct will then be cloned into DNA plasmids optimized for expression in vivo. The vaccine candidates will be systematically examined for their ability to confer therapeutic protection from infection in HLA transgenic mice, and for their ability to avert the development of gastric cancer in a p27-/- mutant mouse model. Results from these studies will identify novel vaccines for control of H. pylori infection, and for the first time, examine directly the protective effect of vaccination against H. pylori-associated malignant disease. Phase I is devoted to feasibility; a Phase II SBIR program would examine the best means of enhancing immunogenicity, using reduction of chronic infection and modulation of metaplastic disease in well-established mouse models of H. pylori disease as our correlate of efficacy.

Keywords: Helicobacter, bacterial antigen, bacterial genetics, bacterial vaccine, vaccine development, vector vaccine, MHC class II antigen, cellular immunity, immune response, neoplasm /cancer pharmacology, plasmid, stomach neoplasm, vaccine evaluation, biotechnology, clinical research, genetically modified animal, human tissue, laboratory mouse

Project start date: 2005-06-01

Project end date: 2008-05-31

5R43AI065036-02 (2006): $309787

Sponsored Links Excellgen http://Excellgen.com

	Recombinant Lentivirus & Adenovirus High Yield and High Titer up to 10¹⁰ (lentivirus) and 10¹³ (adenovirus) for Guaranteed Expression of GOI. ~~$3000~~, $2500
	Baculovirus Protein Expression Fast turn around, >95% purity functional protein. No outsourcing to China or India. ~~$5500~~, $3950
	Transient Protein Expression in CHO and HEK293 Cells Transient Expression, Truly Functional Protein, 95% purity, 1~20 mg, fast turnaround. ~~$5500~~, $3950

1R43AI065036-01 (2005): $300123

Development Of De-immunized Botulinum Neurotoxin Type A For Dystonia

Anne Searls De groot
Epivax, Inc., 146 Clifford Street, Providence, Ri 02903

Grant 1R21NS054781-01A1 from National Institute Of Neurological Disorders And Stroke IRG: NSD

Abstract: DESCRIPTION () This revised Exploratory/Developmental (R21) proposal describes innovative research approaches to eliminate the immune response to botulinum neurotoxin type A (BoNT/A) therapy in dystonia patients. The immunogenicity of BoNT/A gives rise to neutralizing antibodies that reduce or eliminate its therapeutic benefits. The proposed research stems from a recognition that development of protein therapeutics in general must be driven by insights into drug-induced immune responses. A generalizable strategy for de- immunization of functional therapeutics will eliminate or at least reduce this major obstacle to the rapid translation of "genes to drugs". This novel approach combines cutting-edge bioinformatics tools with classic immunological assays to create an improved BoNT/A that escapes immunological detection much in the same way as tumor cells and viruses. The target will be the T cell dependent mechanism for antibody generation, which is mediated by peptide-loaded class II MHC-T cell receptor interactions. The primary goals of the project are to use epitope mapping algorithms to computationally identify BoNT/A- derived T cell epitopes. Subsequent verification will be performed experimentally using MHC binding assays and T cell stimulation studies of blood drawn from BoNT/A exposed patients. Immunodominant epitopes will be targeted for modification to reduce their immunogenicity with the aim of preserving the overall fold of the toxin. Computational methods will be used to verify reduced immunogenicity of the modified toxin, and homology modeling will be used to determine the structural effects of these changes. The de-immunized peptide sequences will be assayed for MHC binding and in vitro and in vivo (mouse) T cell stimulation. Public Health Relatedness Knowledge of the elements of botulinum neurotoxin type A that give rise to its immunogenicity will contribute to the development of improved dystonia therapy.

Project start date: 2007-03-01

Project end date: 2009-02-28

1R21NS054781-01A1 (2007): $153125

Epitope Driven HIV Vaccine Development AI50528-01A1

Anne Searls De groot
Brown University 164 Angell Street Providence, Ri 02912

Grant 5R03TW006306-02 from Fogarty International Center IRG: ZRG1

Abstract: Background The TB/HIV Research Lab s GAlA vaccine is being developed from selected HIV T cell epitopes that are conserved (the same or very similar) across several strains (or clades) of HIV, that are promiscuous (bind to cells from genetically diverse human immune systems), and are likely to elicit both of the major types of T cell immune responses (class I-restricted cytotoxic T cells and class II-restricted T helper cells). Ultimately, these epitopes will be inserted into a DNA plasmid, which will be the primary delivery vector for the vaccine. A "boost" vaccine would contain the same epitopes, in a viral vector or as a protein in adjuvant. NIH R01 "Epitope Driven HIV Vaccine Development" A150528-01A1 provides funds for pre-clinical studies of the GAlA vaccine epitopes using blood from HIV -positive patients recruited in Providence (predominantly clade B infected patients). To confirm the cross-clade immunogenicity of the epitopes, screening in vitro using blood from HIV-positive patients infected with different clades or strains of HIV needs to be performed. Proposal This Fogarty FIRCA proposal would support a collaboration between researchers in Bamako, Mali and the TB/HIV Research Laboratory at Brown University. The goal of this AIDS FIRCA would be to screen the GAlA vaccine epitopes using blood from HIV-infected Malian patients recruited in Bamako, Mali. The GAlA Vaccine / Bamako, Mali project would take place at an existing center of research excellence funded by the NIH (NIAID) at the Faculte de Medicine, University of Mali (The Malaria Research training Center, or MRTC). In the short term, this research would allow the research team to confirm candidate HIV vaccine eptopes in the context of HIV in Mali. In the long term, the goal of this collaboration is to develop a close working relationship between the TB/HIV Research Lab and Malian investigators, who might then host Phase clinical trials of the GAlA vaccine when it is ready.

Keywords: AIDS vaccine, genetic strain, human immunodeficiency virus, immunogenetics, vaccine development, Africa, T lymphocyte, immune response, international cooperation, nucleic acid sequence, virus DNA, clinical research, enzyme linked immunosorbent assay, human subject, patient oriented research, peptide chemical synthesis, peptide library, polymerase chain reaction

Project start date: 2004-07-01

Project end date: 2005-11-30

5R03TW006306-02 (2005): $40320

1R03TW006306-01A2 (2004): $40320

5R03TW006306-04 (2006): $39372

HIV-1/HIV-2 EPITOPE PROJECT, THE GAMBIA, WEST AFRICA

Anne Searls De groot
Brown University
164 Angell Street
providence, Ri 02912

Grant 5R03TW000973-03 from Fogarty International Center IRG: ARRB

Abstract: adapted from the ) In this application, the Principal Investigator outlines an effort to identify "protective" HIV-2 cytotoxic T-cell (CTL) epitopes that are recognized by individuals of Gambian, or sub-Saharan African, genetic backgrounds. The spread of HIV-1 into sub-Saharan countries where HIV-2 was previously endemic has resulted in the exposure of HIV-2 infected individuals to HIV-1. As some HIV2-infected individuals appear to be resistant to HIV-1 infection, some have proposed that recognition of cross-reactive CTL epitopes may contribute to protection from HIV-1. The elucidation of the roles CTL epitopes and genetic background (HLA) play in the resistance to HIV-1 infection in the presence of HIV-2 infection may contribute to our understanding of HIV immunopathogenesis. Advances in the development of computer-driven algorithms that prospectively identify putative MHC-binding regions/CTL epitopes, such as the TB/HIV research laboratory´s EpiMatrix, will facilitate the comparison of immunologically relevant regions of HIV-2 sequences to HIV-1 sequences. The Investigator and her associates have demonstrated that EpiMatrix efficiently and accurately identifies MHC-binding regions from HIV protein sequences. She proposes now to apply the algorithm to the sequences of HIV-2 and HIV-1 strains that are common in West Africa, selecting putative MHC-binding regions that are conserved between these strains. The specific aims of this project are to (1) identify conserved HIV-2/HIV-1 CTL epitopes and (2) test whether these putative epitopes are recognized by HIV-2 infected individuals. The long-term goal is to evaluate the relation between the recognition of the CTL epitopes and presence of HIV-2/HIV-1 co-infection. Modeling and MHC-binding studies will be performed in Providence, Rhode Island (United States) with the participation of MRC personnel. Selected peptides will then be tested in CTL assays using CTL lines and clones derived from HIV-2 infected individuals. The CTL assays will be performed at the Medical Research Council Laboratories in Fajara, The Gambia, West Africa, as a collaboration between TB/HIV Research lab personnel and MRC Laboratories personnel

Keywords: Africa, HIV infection, cytotoxic T lymphocyte, human immunodeficiency virus 1, human immunodeficiency virus 2, virus antigen MHC class I antigen, antigen presentation, mathematical model, protein sequence African, clinical research, human subject

Project start date: 1998-09-01

Project end date: 2002-02-28

5R03TW000973-03 (2001): $40000

5R03TW000973-02 (1999): $40000

TB PEPTIDE EPITOPE PROJECT, MRC LABORATORIES, THE GAMBIA

Anne Searls De groot
Brown University 164 Angell Street Providence, Ri 02912

Grant 5R03TW000552-03 from Fogarty International Center IRG: ARRE

Abstract: Adapted from Applicant s ) Immune responses to M. tuberculosis (MTB) are dependent on the recognition of protective MTB epitopes in the context of the MHC background of the individual at risk. Thus, putative MTB epitopes for a synthetic vaccine against MTB must be tested using T cells from populations at risk for tuberculosis. Tuberculosis is one of the primary manifestation of HIV in The Gambia, a country that bisects Senegal on the West Coast of Africa. Between 80-90 percent of HIV seropositive patients seen at the MRC Fajara Clinic are co-infected with MTB and as many as 20 percent have active tuberculosis. The distribution of MHC alleles in Africa is strikingly different from the distribution of alleles in Caucasian populations. This laboratory group has developed a novel computer-driven algorithm that identifies peptides from MTB protein sequences that contain high densities of MHC binding motifs. Preliminary studies demonstrate that the computer-driven algorithm is both more efficient and more accurate than standard methods of MTB epitope identification. However, many of the evaluations of MTB epitopes have been performed using T cells from predominantly Caucasion donors in developed countries, where tuberculosis is no longer endemic. In this application, novel MTB epitopes will be synthesized that have been predicted by the algorithm and these peptides will be tested in assays to be performed in The Gambia, where tuberculosis is endemic. MHC typing and investigations of the correlation between the MHC type of individuals who respond to the peptides and the motifs contained within the peptide will be performed. The specific aims are 1. To identify "protective" MTB epitopes that are recognized by individuals in a tuberculosis endemic area; 2. To evaluate the predictive power of the computer-driven algoritym for non-Caucasian T cell epitopes; and 3. To evaluate correlations between the MHC motifs included in the predicted peptide and the MHC type of responders. If these novel MHC binding motif-rich epitopes are recognized by MTB-immune individuals in The Gambia, they may be important candidates for inclusion in a synthetic tuberculosis vaccine targeting HIV seropositive individuals in sub Saharan Africa.

Keywords: Africa, Mycobacterium tuberculosis, bacterial antigen, tuberculosis, AIDS, computer program /software, major histocompatibility complex, molecular pathology, peptide chemical synthesis, histocompatibility typing, human subject

Project start date: 1995-09-30

Project end date: 1999-09-29

5R03TW000552-03 (1997): $32000

A CROSS CLADE APPROACH TO HIV VACCINE DEVELOPMENT

Anne Searls De groot
International Health Institutebrown University
164 Angell Street
providence, Ri 02912

Grant 5R21AI045416-02 from National Institute Of Allergy And Infectious Diseases IRG: ZRG1

Abstract: Adapted from Applicant´s ) EpiMer and EpiMatrix are algorithms used to prospectively identify regions of proteins expected to bind to MHC and to be presented to T cells (cytotoxic T cells/CTL and T helper cells/Th). The algorithms have successfully identified novel clade B, C, and E epitopes from primary HIV-1 sequences. These proposed studies will elucidate cross-clade responses to putative HIV-1 T cell epitopes, evaluate the expression and recognition of the epitopes as "cassettes" in DNA plasmids, and explore the utility of including the epitopes in an HIV vaccine. The specific aims of the proposed work are to 1) Identify novel cross-reactive (cross-clade) HIV-1 CTL and Th epitopes using EpiMatrix and 2) Develop plasmids containing these epitopes as inserts, evaluate their expression and recognition in vitro. Regions of HIV that are conserved across HIV strains listed in the Los Alamos HIV Sequence Database have been identified and screened for MHC binding potential using EpiMatrix. An initial set of 25 of the highest scoring peptides that are well conserved across clades has been constructed for each of 32 class 1 alleles; funding to evaluate these putative CTL epitopes in vitro is sought. We also propose to construct a similar list of conserved, high-scoring putative class II-restricted epitopes and to evaluate these epitopes in vitro. Class I-restricted peptides that (1) bind in vitro or (2) receive high estimates of binding probability based on EpiMatrix will be selected for CTL assays to be performed at the TB/HIV Research Laboratory, using T cells derived from HIV-1 (clade B) infected individuals. Class II- restricted peptides will be studied directly in T cell assays (no other surrogate for "epitopicity" exists). Additional T cell assays will be performed by collaborators. T cell epitopes identified in this manner will be expressed as oligonucleotides and inserted in plasmids developed by Dr. Robert Whalen. In year two of the project, assays will be performed to confirm the expression and immunogenicity of the epitopes in the plasmids. These well-conserved HIV-1 CTL and Th epitopes may be useful components of a novel vector-based or plasmid-based DNA vaccine against HIV-1

Keywords: AIDS vaccine, cross immunity, genetic strain, human immunodeficiency virus 1, vaccine development, vector vaccine MHC class I antigen, MHC class II antigen, cytotoxic T lymphocyte, helper T lymphocyte, leukocyte activation /transformation, virus antigen clinical research, human subject

Project start date: 1999-09-30

Project end date: 2001-09-29

5R21AI045416-02 (2000): $215493

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1R21AI045416-01 (1999): $229692

TRIAD PILOT PROJECTS CORE

Anne Searls De groot, Research Professor
University Of Rhode Island, Research Office, Kingston, Ri 02881-0811

Keywords: Immunology; Immunology (Including BRMP); Immunology (NCI Program); Pilot Projects; Research; pilot study; vaccine development

Project start date: 2009-07-20

Project end date: 2014-06-30

Budget start date: 1-APR-2009

Budget end date: 30-JUN-2010

PFA/PA: RFA-AI-08-014

1U19AI082642-01_8120 (2009): $161571

EPITOPE-DRIVEN HIV VACCINE TARGETING DENDRITIC CELLS

Anne Searls De groot
Epivax, Inc.
providence, Ri 02903

Grant 1R43AI046212-01 from National Institute Of Allergy And Infectious Diseases IRG: ZRG1

Keywords: AIDS vaccine, dendritic cell, human immunodeficiency virus 1, vaccine development computer program /software, cytotoxic T lymphocyte, genetic promoter element, major histocompatibility complex, oligonucleotide, protein structure, virus protein clinical research, epitope mapping, human subject, plasmid, transfection vector

Project start date: 1999-09-30

Project end date: 2000-03-31

1R43AI046212-01 (1999): $100000

NATURAL AND PREDICTED ANTIGENS FOR TB IMMUNOTHERAPY

Anne Searls De groot
Medicinebrown University
164 Angell Street
providence, Ri 02912

Grant 5R01AI035271-04 from National Institute Of Allergy And Infectious Diseases IRG: SRC

Project start date: 1993-09-30

Project end date: 1998-06-30

5R01AI035271-04 (1996): $360411

TB PEPTIDE EPITOPE PROJECT, MRC LABORATORIES, THE GAMBIA

Anne Searls De groot
Medicinebrown University
164 Angell Street
providence, Ri 02912

Grant 5R03TW000552-02 from Fogarty International Center IRG: ARRE

Project start date: 1995-09-30

Project end date: 1998-09-29

5R03TW000552-02 (1996): $32000