Vaccine-Induced Immunity to CMV
(2011)
Abstract :
CMV infection of hematopoietic cell transplant (HCT) recipients is a continuing problem that impacts the outcome of this very successful therapy. Anti-viral treatment with ganciclovir/foscarnet is the main treatment strategy to prevent CMV disease post-transplant (Tx). Despite significant advances in formulation and delivery of anti-virals, their use complicates and extends the post-Tx recovery period and risk for CMV disease. Considering these caveats, we are pursuing a novel therapeutic strategy that focuses on priming or enhancing CMV-specific T cell immunity in healthy volunteers and HCT donors. In developing this vaccine strategy, it is assumed that the target population will be immunocompetent, and vaccination judged successful if immunity to CMV in response to the vaccine is equivalent in individuals with self-limited CMV infection, which at first approximation represents a protective response. Towards this end in Specific Aim (SA1) attenuated poxviruses (MVA) will be constructed that express four CMV antigens (CMV-MVA) including UL32, UL44, UL83, and UL123-exon4 for stimulation of cellular immunity to CMV. MVA has several advantages including avirulence, low inflammatory response and pathogenicity in humans, and inclusion of multiple CMV antigens will ensure broad vaccine coverage for the USA. Lack of viral assembly in mammals, together with studies showing its safety in heavily immunosuppressed macaques, rodents, and in HIV-AIDS patients is evidence of its candidacy for use in HCT recipients. CMV-MVA constructed in SA1 will be qualified as immunologically functional, and subsequently transferred to a certified cGMP manufacturer for clinical production. To ensure safety of the rMVA for Tx recipients, a safety study in SA2 will be conducted in healthy CMV positive and negative volunteers. The immunization regimen will model the time frame required for providing two doses of vaccine to Tx donors without delaying Tx. A Phase II trial using a randomized balanced cohort of 140 vaccinees and unvaccinated controls is proposed in SA3 to establish whether immunization of a donor with CMV-MVA prior to Tx will result in modification of the course and magnitude of CMV-viremia in a Tx recipient. This trial will be sufficiently powered that a 62% reduction in viremia will be statistically meaningful as a measure of the success of the vaccine. Successful limitation of acute CMV infection by this approach lays the foundation for addressing the important problem of late CMV disease by immunizing Tx recipients with the same or an improved version of CMV-MVA.
| Project Number : | 2P01CA030206-24A10013 |
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| ICD : | NATIONAL CANCER INSTITUTE |
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| IRG : | ZCA1 |
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| Project Terms : | cytomegalovirus, human therapy evaluation, immunity, vaccine development, viral vaccine Herpesviridae disease, bone marrow transplantation, cytotoxic T lymphocyte, homologous transplantation, immunologic memory, microorganism immunology, neoplasm /cancer transplantation, nonhuman therapy evaluation, virus protein clinical research, epitope mapping, flow cytometry, genetically modified animal, human subject, laboratory mouse, peptide library |
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Peptide Vaccine To Prevent CMV Disease After HSCT
(2010)
Abstract :
DESCRIPTION (provided by applicant): CMV infection of hematopoietic stem cell transplant (HSCT) recipients is a continuing problem that impacts the outcome of this very successful therapy. Anti-viral treatment with ganciclovir/foscarnet is the main treatment strategy to prevent CMV disease post-transplant (Tx). Despite significant advances in formulation and delivery of anti-virals, their use complicates and extends the post-Tx recovery period and risk for CMV disease. The period of immunologic incompetence, pre- and post-Tx complicates the timing of administering immunotherapy. Considering these caveats, we are pursuing a novel therapeutic strategy that focuses on priming or enhancing CMV-specific T cell immunity in healthy adults and HSCT donors. The approach utilizes peptide fusions combining CMV-specific CTL and promiscuous HLA-DR-binding T-help epitopes, whose activity has been demonstrated in transgenic (Tg) HLA A2 mouse models. Clinical studies will focus on the safety and immunogenicity of two candidate peptides manufactured under NCI/RAID auspices. Attaining a frequency of 108/liter CMV-specific CTL in peripheral blood of vaccine recipients is an important quantitative goal that translates into an eventual post-infusion level of (107/liter CMV-specific CTL in HSCT recipients, a level associated with disease protection. A two-stage Phase 1 trial design is proposed in which 1) the safety of two peptide vaccines given at 4 time-points with dose escalation will determine the Maximum Tolerated Dose (MTD) in healthy CMV-positives and negatives and 2) the safety of both peptides with a novel adjuvant, CpG 7909 DNA will be defined. A secondary objective will be to measure immunologic correlates, especially the frequency of CMV-specific CTL following each booster, continuing for one year. The best peptide and adjuvant combination based on immunogenicity to enhance recall and primary immune responses will be selected for further Phase 2 evaluation in HSCT donor-recipient pairs. HSCT donors will receive 3 doses of peptide pre-Tx, followed by a single booster in the recipient, 28d post-Tx. The primary endpoint will be reduction in CMV viremia without ganciclovir treatment. The trial is sufficiently powered to detect a 40% decrease in viremia, distinguishable between vaccines and those offered standard of care. A secondary objective is quantitative measurement of CMV-specific cellular immunity in HSCT recipients for one-year post-Tx. In this trial, protection from CMV disease will be compared between recipients receiving vaccine or standard of care. The goal of developing a universal repertoire of CMV-specific vaccine candidates will be the object of a developmental study of new fusion peptides in Tg mouse models that cover population subsets other than HLA A2 that would be applicable to a majority of HSCT recipients.
| Project Number : | 5R01CA077544-10 |
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| ICD : | NATIONAL CANCER INSTITUTE |
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| IRG : | CONC |
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| Project Terms : | chemoprevention, cytomegalovirus, hematopoietic tissue transplantation, opportunistic infection, stem cell transplantation, synthetic vaccine, vaccine development, vaccine evaluation CpG island, MHC class I antigen, active immunization, clinical trial phase I, clinical trial phase II, cytotoxic T lymphocyte, dosage, drug adverse effect, helper T lymphocyte, immunomodulator, immunopharmacology, immunotherapy, virus disease, virus load biotechnology, clinical research, genetically modified animal, human subject, laboratory mouse, patient oriented research |
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Peptide Vaccine To Prevent CMV Disease After HSCT
(2010)
Abstract :
CMV infection of hematopoietic stem cell transplant (HSCT) recipients is a continuing problem that impacts the outcome of this very successful therapy. Anti-viral treatment with ganciclovir/foscarnet is the main treatment strategy to prevent CMV disease post-transplant (Tx). Despite significant advances in formulation and delivery of anti-virals, their use complicates and extends the post-Tx recovery period and risk for CMV disease. The period of immunologic incompetence, pre- and post-Tx complicates the timing of administering immunotherapy. Considering these caveats, we are pursuing a novel therapeutic strategy that focuses on priming or enhancing CMV-specifie T cell immunity in healthy adults and HSCT donors. The approach utilizes pepfide fusions combining CMV-specific CTL and promiscuous HLA-DR-binding T-help epitopes, whose activity has been demonstrated in transgenic (Tg) HLA A2 mouse models. Clinical studies will focus on the safety and immunogerdcity of two candidate peptides manufactured under NCI/RAID auspices. Attaining a frequency of 10S/liter CMV-specific CTL in peripheral blood of vaccine recipients is an important quantitative goal that translates into an eventual post-infusion level of >107/liter CMV- specific CTL in HSCT recipients, a level associated with disease protection. A two-stage Phase 1 trial design is proposed in which 1) the safety of two pepfide vaccines given at 4 time-points with dose escalation will determine the Maximum Tolerated Dose (MTD) in healthy CMV-positives and negatives and 2) the safety of both peptides with a novel adjuvant, CpG 7909 DNA will be defined. A secondary objective will be to measure immunologic correlates, especially the frequency of CMV-specific CTL following each booster, continuing for one year. The best peptide and adjuvant combination based on immunogenicity to enhance recall and primary immune responses will be selected for further Phase 2 evaluation in HSCT donor-recipient pairs. HSCT donors will receive 3 doses of peptide pre-Tx, followed by a single booster in the recipient, 28d post-Tx. The primary endpoint will be reduction in CMV viremia without ganciclovir treatment. The trial is sufficiently powered to detect a 40% decrease in viremia, distinguishable between vaccinees and those offered standard of care. A secondary objective is quantitative measurement of CMV- specific cellular immunity in HSCT recipients for one-year post-Tx. In this trial, protection from CMV disease will be compared between recipients receiving vaccine or standard of care. The goal of developing a universal repertoire of CMV-specific vaccine candidates will be the object of a developmental study of new fusion peptides in Tg mouse models that cover population subsets other than HLA A2 that would be applicable to a majority of HSCT recipients. PEFORMANCSEITE(S)(organizationc,ity,state) Beckman Research Institute of the City of Hope, Duarte, CA 91010 KEYPERSONNELS. eeinstructionUs.secontinua#opnagesasneededtoprovidetherequireidnformatiointheformasthownbelow. StartwithPrincipaInl vestigatoLri.stallotherkeypersonneinlalphabeticoarlderl,astnamefirst. Name Organization RoteonProject Don J. Diamond, Ph.D. Beckman Research Institute Principal Investigator John A. Zaia, M.D. Beckman Research Institute Co-investigator Ryotaro Nakamura, M.D. City of Hope National Medical Center Co-Investigator Joy Fridey, M.D. City of Hope National Medical Center Co-Investigator Simon F. Lacey, Ph.D. Beckman Research Institute Co-Investigator Cofinna La Rosa, Ph.D. Beckman Research Institute Senior Fellow Jeff Longmate, Ph,D. Beckman Research Institute Bio-statistician Wahajul Haq, Ph.D. Beckman Research Institute Peptide Chemist Stephen J. Forman, M.D. City of Hope National Medical Center Consultant Arthur M. Krieg, M.D. Coley Pharmaceuticals Consultant DisclosurePermissionStatementApplicabletoSBIR/STTROnly.Seeinstructions.[] Yes [] No PHS398(Rev.05/01) Page 2 FormPage2 Principal InvestigatodProgram Director (Last, first, middle): Diamond Don J. The name of the principal investigatodprogram director must be provided at the top of each printed page and each continuation page. RESEARCH GRANT TABLE OF CONTENTS Page Numbers Face Page .................................................................................................................................................. 1 Description,
| Project Number : | 3R01CA077544-10S1 |
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| ICD : | NATIONAL CANCER INSTITUTE |
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| IRG : | CONC |
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Peptide Vaccine To Prevent CMV Disease After HSCT
(2009)
Abstract :
DESCRIPTION (provided by applicant): CMV infection of hematopoietic stem cell transplant (HSCT) recipients is a continuing problem that impacts the outcome of this very successful therapy. Anti-viral treatment with ganciclovir/foscarnet is the main treatment strategy to prevent CMV disease post-transplant (Tx). Despite significant advances in formulation and delivery of anti-virals, their use complicates and extends the post-Tx recovery period and risk for CMV disease. The period of immunologic incompetence, pre- and post-Tx complicates the timing of administering immunotherapy. Considering these caveats, we are pursuing a novel therapeutic strategy that focuses on priming or enhancing CMV-specific T cell immunity in healthy adults and HSCT donors. The approach utilizes peptide fusions combining CMV-specific CTL and promiscuous HLA-DR-binding T-help epitopes, whose activity has been demonstrated in transgenic (Tg) HLA A2 mouse models. Clinical studies will focus on the safety and immunogenicity of two candidate peptides manufactured under NCI/RAID auspices. Attaining a frequency of 108/liter CMV-specific CTL in peripheral blood of vaccine recipients is an important quantitative goal that translates into an eventual post-infusion level of (107/liter CMV-specific CTL in HSCT recipients, a level associated with disease protection. A two-stage Phase 1 trial design is proposed in which 1) the safety of two peptide vaccines given at 4 time-points with dose escalation will determine the Maximum Tolerated Dose (MTD) in healthy CMV-positives and negatives and 2) the safety of both peptides with a novel adjuvant, CpG 7909 DNA will be defined. A secondary objective will be to measure immunologic correlates, especially the frequency of CMV-specific CTL following each booster, continuing for one year. The best peptide and adjuvant combination based on immunogenicity to enhance recall and primary immune responses will be selected for further Phase 2 evaluation in HSCT donor-recipient pairs. HSCT donors will receive 3 doses of peptide pre-Tx, followed by a single booster in the recipient, 28d post-Tx. The primary endpoint will be reduction in CMV viremia without ganciclovir treatment. The trial is sufficiently powered to detect a 40% decrease in viremia, distinguishable between vaccines and those offered standard of care. A secondary objective is quantitative measurement of CMV-specific cellular immunity in HSCT recipients for one-year post-Tx. In this trial, protection from CMV disease will be compared between recipients receiving vaccine or standard of care. The goal of developing a universal repertoire of CMV-specific vaccine candidates will be the object of a developmental study of new fusion peptides in Tg mouse models that cover population subsets other than HLA A2 that would be applicable to a majority of HSCT recipients.
| Project Number : | 3R01CA077544-07S1 |
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| ICD : | NATIONAL CANCER INSTITUTE |
|---|
| IRG : | CONC |
|---|
| Project Terms : | chemoprevention, cytomegalovirus, hematopoietic tissue transplantation, opportunistic infection, stem cell transplantation, synthetic vaccine, vaccine development, vaccine evaluation CpG island, MHC class I antigen, active immunization, clinical trial phase I, clinical trial phase II, cytotoxic T lymphocyte, dosage, drug adverse effect, helper T lymphocyte, immunomodulator, immunopharmacology, immunotherapy, viremia, virus disease biotechnology, clinical research, genetically modified animal, human subject, laboratory mouse, patient oriented research |
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Peptide Vaccine To Prevent CMV Disease After HSCT
(2009)
Abstract :
DESCRIPTION (provided by applicant): CMV infection of hematopoietic stem cell transplant (HSCT) recipients is a continuing problem that impacts the outcome of this very successful therapy. Anti-viral treatment with ganciclovir/foscarnet is the main treatment strategy to prevent CMV disease post-transplant (Tx). Despite significant advances in formulation and delivery of anti-virals, their use complicates and extends the post-Tx recovery period and risk for CMV disease. The period of immunologic incompetence, pre- and post-Tx complicates the timing of administering immunotherapy. Considering these caveats, we are pursuing a novel therapeutic strategy that focuses on priming or enhancing CMV-specific T cell immunity in healthy adults and HSCT donors. The approach utilizes peptide fusions combining CMV-specific CTL and promiscuous HLA-DR-binding T-help epitopes, whose activity has been demonstrated in transgenic (Tg) HLA A2 mouse models. Clinical studies will focus on the safety and immunogenicity of two candidate peptides manufactured under NCI/RAID auspices. Attaining a frequency of 108/liter CMV-specific CTL in peripheral blood of vaccine recipients is an important quantitative goal that translates into an eventual post-infusion level of (107/liter CMV-specific CTL in HSCT recipients, a level associated with disease protection. A two-stage Phase 1 trial design is proposed in which 1) the safety of two peptide vaccines given at 4 time-points with dose escalation will determine the Maximum Tolerated Dose (MTD) in healthy CMV-positives and negatives and 2) the safety of both peptides with a novel adjuvant, CpG 7909 DNA will be defined. A secondary objective will be to measure immunologic correlates, especially the frequency of CMV-specific CTL following each booster, continuing for one year. The best peptide and adjuvant combination based on immunogenicity to enhance recall and primary immune responses will be selected for further Phase 2 evaluation in HSCT donor-recipient pairs. HSCT donors will receive 3 doses of peptide pre-Tx, followed by a single booster in the recipient, 28d post-Tx. The primary endpoint will be reduction in CMV viremia without ganciclovir treatment. The trial is sufficiently powered to detect a 40% decrease in viremia, distinguishable between vaccines and those offered standard of care. A secondary objective is quantitative measurement of CMV-specific cellular immunity in HSCT recipients for one-year post-Tx. In this trial, protection from CMV disease will be compared between recipients receiving vaccine or standard of care. The goal of developing a universal repertoire of CMV-specific vaccine candidates will be the object of a developmental study of new fusion peptides in Tg mouse models that cover population subsets other than HLA A2 that would be applicable to a majority of HSCT recipients.
| Project Number : | 5R01CA077544-07 |
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| ICD : | NATIONAL CANCER INSTITUTE |
|---|
| IRG : | CONC |
|---|
| Project Terms : | chemoprevention, cytomegalovirus, hematopoietic tissue transplantation, opportunistic infection, stem cell transplantation, synthetic vaccine, vaccine development, vaccine evaluation CpG island, MHC class I antigen, active immunization, clinical trial phase I, clinical trial phase II, cytotoxic T lymphocyte, dosage, drug adverse effect, helper T lymphocyte, immunomodulator, immunopharmacology, immunotherapy, viremia, virus disease biotechnology, clinical research, genetically modified animal, human subject, laboratory mouse, patient oriented research |
|---|
Peptide Vaccine To Prevent CMV Disease After HSCT
(2009)
Abstract :
DESCRIPTION (provided by applicant): CMV infection of hematopoietic stem cell transplant (HSCT) recipients is a continuing problem that impacts the outcome of this very successful therapy. Anti-viral treatment with ganciclovir/foscarnet is the main treatment strategy to prevent CMV disease post-transplant (Tx). Despite significant advances in formulation and delivery of anti-virals, their use complicates and extends the post-Tx recovery period and risk for CMV disease. The period of immunologic incompetence, pre- and post-Tx complicates the timing of administering immunotherapy. Considering these caveats, we are pursuing a novel therapeutic strategy that focuses on priming or enhancing CMV-specific T cell immunity in healthy adults and HSCT donors. The approach utilizes peptide fusions combining CMV-specific CTL and promiscuous HLA-DR-binding T-help epitopes, whose activity has been demonstrated in transgenic (Tg) HLA A2 mouse models. Clinical studies will focus on the safety and immunogenicity of two candidate peptides manufactured under NCI/RAID auspices. Attaining a frequency of 108/liter CMV-specific CTL in peripheral blood of vaccine recipients is an important quantitative goal that translates into an eventual post-infusion level of (107/liter CMV-specific CTL in HSCT recipients, a level associated with disease protection. A two-stage Phase 1 trial design is proposed in which 1) the safety of two peptide vaccines given at 4 time-points with dose escalation will determine the Maximum Tolerated Dose (MTD) in healthy CMV-positives and negatives and 2) the safety of both peptides with a novel adjuvant, CpG 7909 DNA will be defined. A secondary objective will be to measure immunologic correlates, especially the frequency of CMV-specific CTL following each booster, continuing for one year. The best peptide and adjuvant combination based on immunogenicity to enhance recall and primary immune responses will be selected for further Phase 2 evaluation in HSCT donor-recipient pairs. HSCT donors will receive 3 doses of peptide pre-Tx, followed by a single booster in the recipient, 28d post-Tx. The primary endpoint will be reduction in CMV viremia without ganciclovir treatment. The trial is sufficiently powered to detect a 40% decrease in viremia, distinguishable between vaccines and those offered standard of care. A secondary objective is quantitative measurement of CMV-specific cellular immunity in HSCT recipients for one-year post-Tx. In this trial, protection from CMV disease will be compared between recipients receiving vaccine or standard of care. The goal of developing a universal repertoire of CMV-specific vaccine candidates will be the object of a developmental study of new fusion peptides in Tg mouse models that cover population subsets other than HLA A2 that would be applicable to a majority of HSCT recipients.
| Project Number : | 5R01CA077544-09 |
|---|
| ICD : | NATIONAL CANCER INSTITUTE |
|---|
| IRG : | CONC |
|---|
| Project Terms : | chemoprevention, cytomegalovirus, hematopoietic tissue transplantation, opportunistic infection, stem cell transplantation, synthetic vaccine, vaccine development, vaccine evaluation CpG island, MHC class I antigen, active immunization, clinical trial phase I, clinical trial phase II, cytotoxic T lymphocyte, dosage, drug adverse effect, helper T lymphocyte, immunomodulator, immunopharmacology, immunotherapy, virus disease, virus load biotechnology, clinical research, genetically modified animal, human subject, laboratory mouse, patient oriented research |
|---|
Peptide Vaccine To Prevent CMV Disease After HSCT
(2009)
Abstract :
DESCRIPTION (provided by applicant): CMV infection of hematopoietic stem cell transplant (HSCT) recipients is a continuing problem that impacts the outcome of this very successful therapy. Anti-viral treatment with ganciclovir/foscarnet is the main treatment strategy to prevent CMV disease post-transplant (Tx). Despite significant advances in formulation and delivery of anti-virals, their use complicates and extends the post-Tx recovery period and risk for CMV disease. The period of immunologic incompetence, pre- and post-Tx complicates the timing of administering immunotherapy. Considering these caveats, we are pursuing a novel therapeutic strategy that focuses on priming or enhancing CMV-specific T cell immunity in healthy adults and HSCT donors. The approach utilizes peptide fusions combining CMV-specific CTL and promiscuous HLA-DR-binding T-help epitopes, whose activity has been demonstrated in transgenic (Tg) HLA A2 mouse models. Clinical studies will focus on the safety and immunogenicity of two candidate peptides manufactured under NCI/RAID auspices. Attaining a frequency of 108/liter CMV-specific CTL in peripheral blood of vaccine recipients is an important quantitative goal that translates into an eventual post-infusion level of (107/liter CMV-specific CTL in HSCT recipients, a level associated with disease protection. A two-stage Phase 1 trial design is proposed in which 1) the safety of two peptide vaccines given at 4 time-points with dose escalation will determine the Maximum Tolerated Dose (MTD) in healthy CMV-positives and negatives and 2) the safety of both peptides with a novel adjuvant, CpG 7909 DNA will be defined. A secondary objective will be to measure immunologic correlates, especially the frequency of CMV-specific CTL following each booster, continuing for one year. The best peptide and adjuvant combination based on immunogenicity to enhance recall and primary immune responses will be selected for further Phase 2 evaluation in HSCT donor-recipient pairs. HSCT donors will receive 3 doses of peptide pre-Tx, followed by a single booster in the recipient, 28d post-Tx. The primary endpoint will be reduction in CMV viremia without ganciclovir treatment. The trial is sufficiently powered to detect a 40% decrease in viremia, distinguishable between vaccines and those offered standard of care. A secondary objective is quantitative measurement of CMV-specific cellular immunity in HSCT recipients for one-year post-Tx. In this trial, protection from CMV disease will be compared between recipients receiving vaccine or standard of care. The goal of developing a universal repertoire of CMV-specific vaccine candidates will be the object of a developmental study of new fusion peptides in Tg mouse models that cover population subsets other than HLA A2 that would be applicable to a majority of HSCT recipients.
| Project Number : | 3R01CA077544-08S1 |
|---|
| ICD : | NATIONAL CANCER INSTITUTE |
|---|
| IRG : | CONC |
|---|
| Project Terms : | chemoprevention, cytomegalovirus, hematopoietic tissue transplantation, opportunistic infection, stem cell transplantation, synthetic vaccine, vaccine development, vaccine evaluation CpG island, MHC class I antigen, active immunization, clinical trial phase I, clinical trial phase II, cytotoxic T lymphocyte, dosage, drug adverse effect, helper T lymphocyte, immunomodulator, immunopharmacology, immunotherapy, virus disease, virus load biotechnology, clinical research, genetically modified animal, human subject, laboratory mouse, patient oriented research |
|---|
Peptide Vaccine To Prevent CMV Disease After HSCT
(2009)
| Project Number : | 3R01CA077544-09S1 |
|---|
| ICD : | NATIONAL CANCER INSTITUTE |
|---|
| IRG : | CONC |
|---|
Peptide Vaccine To Prevent CMV Disease After HSCT
(2009)
Abstract :
DESCRIPTION (provided by applicant): CMV infection of hematopoietic stem cell transplant (HSCT) recipients is a continuing problem that impacts the outcome of this very successful therapy. Anti-viral treatment with ganciclovir/foscarnet is the main treatment strategy to prevent CMV disease post-transplant (Tx). Despite significant advances in formulation and delivery of anti-virals, their use complicates and extends the post-Tx recovery period and risk for CMV disease. The period of immunologic incompetence, pre- and post-Tx complicates the timing of administering immunotherapy. Considering these caveats, we are pursuing a novel therapeutic strategy that focuses on priming or enhancing CMV-specific T cell immunity in healthy adults and HSCT donors. The approach utilizes peptide fusions combining CMV-specific CTL and promiscuous HLA-DR-binding T-help epitopes, whose activity has been demonstrated in transgenic (Tg) HLA A2 mouse models. Clinical studies will focus on the safety and immunogenicity of two candidate peptides manufactured under NCI/RAID auspices. Attaining a frequency of 108/liter CMV-specific CTL in peripheral blood of vaccine recipients is an important quantitative goal that translates into an eventual post-infusion level of (107/liter CMV-specific CTL in HSCT recipients, a level associated with disease protection. A two-stage Phase 1 trial design is proposed in which 1) the safety of two peptide vaccines given at 4 time-points with dose escalation will determine the Maximum Tolerated Dose (MTD) in healthy CMV-positives and negatives and 2) the safety of both peptides with a novel adjuvant, CpG 7909 DNA will be defined. A secondary objective will be to measure immunologic correlates, especially the frequency of CMV-specific CTL following each booster, continuing for one year. The best peptide and adjuvant combination based on immunogenicity to enhance recall and primary immune responses will be selected for further Phase 2 evaluation in HSCT donor-recipient pairs. HSCT donors will receive 3 doses of peptide pre-Tx, followed by a single booster in the recipient, 28d post-Tx. The primary endpoint will be reduction in CMV viremia without ganciclovir treatment. The trial is sufficiently powered to detect a 40% decrease in viremia, distinguishable between vaccines and those offered standard of care. A secondary objective is quantitative measurement of CMV-specific cellular immunity in HSCT recipients for one-year post-Tx. In this trial, protection from CMV disease will be compared between recipients receiving vaccine or standard of care. The goal of developing a universal repertoire of CMV-specific vaccine candidates will be the object of a developmental study of new fusion peptides in Tg mouse models that cover population subsets other than HLA A2 that would be applicable to a majority of HSCT recipients.
| Project Number : | 5R01CA077544-08 |
|---|
| ICD : | NATIONAL CANCER INSTITUTE |
|---|
| IRG : | CONC |
|---|
| Project Terms : | chemoprevention, cytomegalovirus, hematopoietic tissue transplantation, opportunistic infection, stem cell transplantation, synthetic vaccine, vaccine development, vaccine evaluation CpG island, MHC class I antigen, active immunization, clinical trial phase I, clinical trial phase II, cytotoxic T lymphocyte, dosage, drug adverse effect, helper T lymphocyte, immunomodulator, immunopharmacology, immunotherapy, virus disease, virus load biotechnology, clinical research, genetically modified animal, human subject, laboratory mouse, patient oriented research |
|---|
Vaccine-Induced Immunity to CMV
(2009)
Abstract :
CMV infection of hematopoietic cell transplant (HCT) recipients is a continuing problem that impacts the outcome of this very successful therapy. Anti-viral treatment with ganciclovir/foscarnet is the main treatment strategy to prevent CMV disease post-transplant (Tx). Despite significant advances in formulation and delivery of anti-virals, their use complicates and extends the post-Tx recovery period and risk for CMV disease. Considering these caveats, we are pursuing a novel therapeutic strategy that focuses on priming or enhancing CMV-specific T cell immunity in healthy volunteers and HCT donors. In developing this vaccine strategy, it is assumed that the target population will be immunocompetent, and vaccination judged successful if immunity to CMV in response to the vaccine is equivalent in individuals with self-limited CMV infection, which at first approximation represents a protective response. Towards this end in Specific Aim (SA1) attenuated poxviruses (MVA) will be constructed that express four CMV antigens (CMV-MVA) including UL32, UL44, UL83, and UL123-exon4 for stimulation of cellular immunity to CMV. MVA has several advantages including avirulence, low inflammatory response and pathogenicity in humans, and inclusion of multiple CMV antigens will ensure broad vaccine coverage for the USA. Lack of viral assembly in mammals, together with studies showing its safety in heavily immunosuppressed macaques, rodents, and in HIV-AIDS patients is evidence of its candidacy for use in HCT recipients. CMV-MVA constructed in SA1 will be qualified as immunologically functional, and subsequently transferred to a certified cGMP manufacturer for clinical production. To ensure safety of the rMVA for Tx recipients, a safety study in SA2 will be conducted in healthy CMV positive and negative volunteers. The immunization regimen will model the time frame required for providing two doses of vaccine to Tx donors without delaying Tx. A Phase II trial using a randomized balanced cohort of 140 vaccinees and unvaccinated controls is proposed in SA3 to establish whether immunization of a donor with CMV-MVA prior to Tx will result in modification of the course and magnitude of CMV-viremia in a Tx recipient. This trial will be sufficiently powered that a 62% reduction in viremia will be statistically meaningful as a measure of the success of the vaccine. Successful limitation of acute CMV infection by this approach lays the foundation for addressing the important problem of late CMV disease by immunizing Tx recipients with the same or an improved version of CMV-MVA.
| Project Number : | 5P01CA030206-270013 |
|---|
| ICD : | NATIONAL CANCER INSTITUTE |
|---|
| IRG : | ZCA1 |
|---|
| Project Terms : | cytomegalovirus, human therapy evaluation, immunity, vaccine development, viral vaccine Herpesviridae disease, bone marrow transplantation, cytotoxic T lymphocyte, homologous transplantation, immunologic memory, microorganism immunology, neoplasm /cancer transplantation, nonhuman therapy evaluation, virus protein clinical research, epitope mapping, flow cytometry, genetically modified animal, human subject, laboratory mouse, peptide library |
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CpG DNA:Epitope Fusion Peptide Conjugates as HIV Vaccine
(2008)
Abstract :
DESCRIPTION (provided by applicant): An effective vaccine against HIV infection remains an elusive goal, especially as the sequence diversity of the virus continues to expand. We developed a model system that relies on the recognition of T cell epitopes from immunodominant antigens of HIV to explore novel uses of oligodeoxynucleotides (ODN) as adjuvants, with the purpose of developing an improved strategy for vaccine delivery. We increased the sensitivity of our approach, by developing a novel synthetic chemical strategy of covalently modifying T-help/CTL epitope fusion peptides with CpG-ODN motifs. In this revised application, specific concerns of the Study Section have been addressed in preliminary data including the necessity for a T-help epitope, the fate of epitopes in the middle of peptide strings, and the evaluation of human-specific CpG-ODN. Immunogenicity, in vitro recognition of HIV-infected T cells, and protection against viral infection using a surrogate virus (poxvirus) will be criteria to measure efficacy of newly derived CpG-ODN:peptide conjugates in HLA transgenic murine models. The focus of the experimental section will be on strategies that yield greater sensitivity of immune stimulation by the conjugate vaccines, as well as broadness and strength of immune responses to epitopes from multiple antigens. The methodology for conjugating peptide to CpG-ODN will be intensively investigated in Specific Aim 1, with the purpose of disclosing new chemical structures that will heighten the sensitivity of immune response to the vaccine. A more focused ranking formula that distinguishes vaccines targeting acute and chronic HIV infection will be instituted. Since HIV has immense capacity for escape from cellular immunity, we will seek to combine epitopes from multiple antigens to prevent escape from immune recognition by HIV. In Specific Aim 2, we will explore protection models to assess whether the synthetic vaccine strategies that are most immunogenic in Specific Aim 1, also are more efficient at providing protection against an HIV-antigen recombinant poxvirus challenge. These will include more virulent forms of vaccinia virus, whose clearance may closely model outcomes of human viral infection with un-attenuated viruses. In addition, we will explore whether increased efficiency of immune recognition by CpG-ODN:peptide conjugates will also apply to HIV strains including clades A-D in direct lytic assays of HIV-infected Jurkat T cells. These studies will establish feasibility and provide impetus to extend these observations into primate models of HIV infection, and will justify the intensive studies proposed in this application.
| Project Number : | 5R21AI062496-02 |
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| ICD : | NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES |
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| IRG : | VACC |
|---|
| Project Terms : | AIDS vaccine, HIV infection, chemical structure function, immunoconjugate, vaccine development, vaccine evaluation CpG island, Poxviridae, gene expression, helper T lymphocyte, host organism interaction, oligonucleotide, synthetic peptide, vaccinia virus biotechnology, flow cytometry, genetically modified animal, laboratory mouse |
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Vaccine-Induced Immunity to CMV
(2008)
Abstract :
CMV infection of hematopoietic cell transplant (HCT) recipients is a continuing problem that impacts the outcome of this very successful therapy. Anti-viral treatment with ganciclovir/foscarnet is the main treatment strategy to prevent CMV disease post-transplant (Tx). Despite significant advances in formulation and delivery of anti-virals, their use complicates and extends the post-Tx recovery period and risk for CMV disease. Considering these caveats, we are pursuing a novel therapeutic strategy that focuses on priming or enhancing CMV-specific T cell immunity in healthy volunteers and HCT donors. In developing this vaccine strategy, it is assumed that the target population will be immunocompetent, and vaccination judged successful if immunity to CMV in response to the vaccine is equivalent in individuals with self-limited CMV infection, which at first approximation represents a protective response. Towards this end in Specific Aim (SA1) attenuated poxviruses (MVA) will be constructed that express four CMV antigens (CMV-MVA) including UL32, UL44, UL83, and UL123-exon4 for stimulation of cellular immunity to CMV. MVA has several advantages including avirulence, low inflammatory response and pathogenicity in humans, and inclusion of multiple CMV antigens will ensure broad vaccine coverage for the USA. Lack of viral assembly in mammals, together with studies showing its safety in heavily immunosuppressed macaques, rodents, and in HIV-AIDS patients is evidence of its candidacy for use in HCT recipients. CMV-MVA constructed in SA1 will be qualified as immunologically functional, and subsequently transferred to a certified cGMP manufacturer for clinical production. To ensure safety of the rMVA for Tx recipients, a safety study in SA2 will be conducted in healthy CMV positive and negative volunteers. The immunization regimen will model the time frame required for providing two doses of vaccine to Tx donors without delaying Tx. A Phase II trial using a randomized balanced cohort of 140 vaccinees and unvaccinated controls is proposed in SA3 to establish whether immunization of a donor with CMV-MVA prior to Tx will result in modification of the course and magnitude of CMV-viremia in a Tx recipient. This trial will be sufficiently powered that a 62% reduction in viremia will be statistically meaningful as a measure of the success of the vaccine. Successful limitation of acute CMV infection by this approach lays the foundation for addressing the important problem of late CMV disease by immunizing Tx recipients with the same or an improved version of CMV-MVA.
| Project Number : | 5P01CA030206-260013 |
|---|
| ICD : | NATIONAL CANCER INSTITUTE |
|---|
| IRG : | ZCA1 |
|---|
| Project Terms : | cytomegalovirus, human therapy evaluation, immunity, vaccine development, viral vaccine Herpesviridae disease, bone marrow transplantation, cytotoxic T lymphocyte, homologous transplantation, immunologic memory, microorganism immunology, neoplasm /cancer transplantation, nonhuman therapy evaluation, virus protein clinical research, epitope mapping, flow cytometry, genetically modified animal, human subject, laboratory mouse, peptide library |
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CpG DNA:Epitope Fusion Peptide Conjugates as HIV Vaccine
(2007)
Abstract :
DESCRIPTION (provided by applicant): An effective vaccine against HIV infection remains an elusive goal, especially as the sequence diversity of the virus continues to expand. We developed a model system that relies on the recognition of T cell epitopes from immunodominant antigens of HIV to explore novel uses of oligodeoxynucleotides (ODN) as adjuvants, with the purpose of developing an improved strategy for vaccine delivery. We increased the sensitivity of our approach, by developing a novel synthetic chemical strategy of covalently modifying T-help/CTL epitope fusion peptides with CpG-ODN motifs. In this revised application, specific concerns of the Study Section have been addressed in preliminary data including the necessity for a T-help epitope, the fate of epitopes in the middle of peptide strings, and the evaluation of human-specific CpG-ODN. Immunogenicity, in vitro recognition of HIV-infected T cells, and protection against viral infection using a surrogate virus (poxvirus) will be criteria to measure efficacy of newly derived CpG-ODN:peptide conjugates in HLA transgenic murine models. The focus of the experimental section will be on strategies that yield greater sensitivity of immune stimulation by the conjugate vaccines, as well as broadness and strength of immune responses to epitopes from multiple antigens. The methodology for conjugating peptide to CpG-ODN will be intensively investigated in Specific Aim 1, with the purpose of disclosing new chemical structures that will heighten the sensitivity of immune response to the vaccine. A more focused ranking formula that distinguishes vaccines targeting acute and chronic HIV infection will be instituted. Since HIV has immense capacity for escape from cellular immunity, we will seek to combine epitopes from multiple antigens to prevent escape from immune recognition by HIV. In Specific Aim 2, we will explore protection models to assess whether the synthetic vaccine strategies that are most immunogenic in Specific Aim 1, also are more efficient at providing protection against an HIV-antigen recombinant poxvirus challenge. These will include more virulent forms of vaccinia virus, whose clearance may closely model outcomes of human viral infection with un-attenuated viruses. In addition, we will explore whether increased efficiency of immune recognition by CpG-ODN:peptide conjugates will also apply to HIV strains including clades A-D in direct lytic assays of HIV-infected Jurkat T cells. These studies will establish feasibility and provide impetus to extend these observations into primate models of HIV infection, and will justify the intensive studies proposed in this application.
| Project Number : | 1R21AI062496-01A1 |
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| ICD : | NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES |
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| IRG : | VACC |
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| Project Terms : | AIDS vaccine, HIV infection, chemical structure function, immunoconjugate, vaccine development, vaccine evaluation CpG island, Poxviridae, gene expression, helper T lymphocyte, host organism interaction, oligonucleotide, synthetic peptide, vaccinia virus biotechnology, flow cytometry, genetically modified animal, laboratory mouse |
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Vaccine-Induced Immunity to CMV
(2007)
Abstract :
CMV infection of hematopoietic cell transplant (HCT) recipients is a continuing problem that impacts the outcome of this very successful therapy. Anti-viral treatment with ganciclovir/foscarnet is the main treatment strategy to prevent CMV disease post-transplant (Tx). Despite significant advances in formulation and delivery of anti-virals, their use complicates and extends the post-Tx recovery period and risk for CMV disease. Considering these caveats, we are pursuing a novel therapeutic strategy that focuses on priming or enhancing CMV-specific T cell immunity in healthy volunteers and HCT donors. In developing this vaccine strategy, it is assumed that the target population will be immunocompetent, and vaccination judged successful if immunity to CMV in response to the vaccine is equivalent in individuals with self-limited CMV infection, which at first approximation represents a protective response. Towards this end in Specific Aim (SA1) attenuated poxviruses (MVA) will be constructed that express four CMV antigens (CMV-MVA) including UL32, UL44, UL83, and UL123-exon4 for stimulation of cellular immunity to CMV. MVA has several advantages including avirulence, low inflammatory response and pathogenicity in humans, and inclusion of multiple CMV antigens will ensure broad vaccine coverage for the USA. Lack of viral assembly in mammals, together with studies showing its safety in heavily immunosuppressed macaques, rodents, and in HIV-AIDS patients is evidence of its candidacy for use in HCT recipients. CMV-MVA constructed in SA1 will be qualified as immunologically functional, and subsequently transferred to a certified cGMP manufacturer for clinical production. To ensure safety of the rMVA for Tx recipients, a safety study in SA2 will be conducted in healthy CMV positive and negative volunteers. The immunization regimen will model the time frame required for providing two doses of vaccine to Tx donors without delaying Tx. A Phase II trial using a randomized balanced cohort of 140 vaccinees and unvaccinated controls is proposed in SA3 to establish whether immunization of a donor with CMV-MVA prior to Tx will result in modification of the course and magnitude of CMV-viremia in a Tx recipient. This trial will be sufficiently powered that a 62% reduction in viremia will be statistically meaningful as a measure of the success of the vaccine. Successful limitation of acute CMV infection by this approach lays the foundation for addressing the important problem of late CMV disease by immunizing Tx recipients with the same or an improved version of CMV-MVA.
| Project Number : | 5P01CA030206-250013 |
|---|
| ICD : | NATIONAL CANCER INSTITUTE |
|---|
| IRG : | ZCA1 |
|---|
| Project Terms : | cytomegalovirus, human therapy evaluation, immunity, vaccine development, viral vaccine Herpesviridae disease, bone marrow transplantation, cytotoxic T lymphocyte, homologous transplantation, immunologic memory, microorganism immunology, neoplasm /cancer transplantation, nonhuman therapy evaluation, virus protein clinical research, epitope mapping, flow cytometry, genetically modified animal, human subject, laboratory mouse, peptide library |
|---|
LIPOPEPTIDE VACCINE TO PREVENT HCMV INFECTION AFTER BMT
(2004)
Abstract :
DESCRIPTION: (Applicant's Abstract) A complication of the recovery phase after BMT is cytomegalovirus (HCMV) infection. Therapy using gancyclovir prophylaxis has decreased the mortality from HCMV infection in allogeneic BMT recipients. However side-effects which include delayed immune reconstitution limit its effectiveness in reducing overall mortality from BMT. Studies from several laboratories have indicated that control of CMV infection in BMT recipients is dependent on a strong cytotoxic T lymphocyte (CTL) response which is targeted mainly against the viral matrix protein pp65, and to a lesser degree against pp150. An adoptive immunotherapy strategy using CTL has proved successful but is impractical for general use in BMT centers. The applicant will develop an alternative strategy that uses small peptide fragments from the CTL viral target proteins pp65 and pp150 in the form of a vaccine as a means to simulate CTL in vivo. The vaccine is a peptide composed of a CTL epitope combined with a peptide which stimulates a T helper (TH) response and is lipidated on the amino terminus. He will first evaluate several different structural forms of the vaccine containing the HLA-A0201 restricted CTL epitope from pp65 and several different TH epitopes in a transgenic HLA-A2.1 mouse model. He will evaluate which of the vaccine molecules provide the strongest and longest-lived immunity by comparing the degree of lysis in vitro of human fibroblasts infected with HCMV mediated by murine splenic effector CTL. Based on the results of murine immunizations, a Phase I dose-escalating and safety trial will be conducted in asymptomatic HCMV seropositive human volunteers with a lipidated HLA-A0201 restricted CTL vaccine. Additional vaccines including those with 2 or 3 CTL epitopes will also be evaluated in human volunteers in a Phase I trial. In order to test whether the vaccine, which enhances CTL against HCMV in healthy volunteers, is also capable of protecting against infection, a Phase II trial will be conducted in HCMV seropositive BMT donors and recipients. BMT donors will be vaccinated according to a protocol determined by the Phase I trial and their HLA-matched siblings who undergo BMT will be monitored for the ability to resist viremia and/or pneumonia brought about by HCMV infection. Success of this protocol will be determined by the avoidance of gancyclovir post-BMT and reduction of early and late HCMV infection in the BMT recipient.
| Project Number : | 5R01CA077544-05 |
|---|
| ICD : | NATIONAL CANCER INSTITUTE |
|---|
| IRG : | ET |
|---|
| Project Terms : | bone marrow transplantation, chemoprevention, cytomegalovirus, drug screening /evaluation, postoperative complication, synthetic vaccine, viral vaccine, virus infection mechanism MHC class I antigen, active immunization, clinical trial phase I, clinical trial phase II /III /IV, cytotoxic T lymphocyte, dosage, drug adverse effect, extracellular matrix protein, helper T lymphocyte, viremia, virus protein clinical research, human subject, laboratory mouse, lung lavage, polymerase chain reaction, transgenic animal |
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ACQUISITION OF IMMUNITY TO CYTOMEGALOVIRUS
(2004)
Abstract :
CMV infection remains an important problem for transplantation. No successful vaccine has been developed that prevents new infections or even controls existing ones. Pharmacologic treatments have limitations regarding their side effects and the development of resistant strain Asymptomatic CMV-seropositive individuals of survivors of CMV infection after BMT, have an ongoing cytotoxic T lymphocyte (CTL) response to CMV, which can be measured after in vitro stimulation (IVS), either using HLA tetrameter binding or chromium release assay (CRA). The CTL response has multiple targets, however, the response to the tegument protein pp65 predominates in most individuals examined. Although many CTL epitopes have already been mapped from two immunodominant CMV proteins (pp65 and pp150), a significant number have yet been determined, and their elucidation will lead to a broad-based vaccine strategy for all major ethic groups in the United States. Immunodominance of CTL responses to CMV proteins will be determined utilizing IVS strategies together with known CTL epitopes. In contrast, little is known of the predominance of CTL targets from CMV in BMT recipients. The frequency of CMV-specific CTL will be enumerated using flow cytometry with HLA tetramers, and the frequency of those CTL will be compared to viral load measurements using plasma PCR methods. As a means to enhance the immunogenicity of previous defined CTL epitopes, a combinatorial peptide chemistry approach will be used with the known CTL epitopes from pp65 and pp150, which have affinities that are in the micromolar to approach will be used with the known CTL epitopes from pp65 and pp150, which have affinities that are in the micromolar to nanomolar range. Analogue peptides will be defined that will be evaluated initially using HLA A*0201 restricted and pp65- specific T cell clones, to develop a more immunogenic CTL epitope which may lead to a more effective CMV vaccine. The functionality of these analogue peptides will be shown utilizing in vivo immunization of HLA-transgenic mice and IVS studies stimulating a memory response in human PBMC specific for killing of CMV infected fibroblasts in vitro. Two Phase II trials will evaluate modalities including CMV lipopeptide immunization in combination with standard tetanus toxin immunization of BMT recipients. Augmentation of the memory response to tetanus- specific CD4+ T cells will be evaluated by in vitro methods, and the duration of infection-free survival of BMT recipients will be measured using standard clinical parameters. Finally, the utility of targeting a single CMV protein utilizing a CTL epitope vaccine strategy versus targeting two or more CMV proteins will be evaluated using CMV lipopeptide immunization of BMT recipients. Since the endpoint of the trial is a reduction in the incidence of CMV-associated disease, the comparison must be done utilizing a patient population likely to have measurable CMV viral load. Taken together, these strategies of defining CTL epitopes, augmenting their immunogenicity, and defining efficacious ways to delay them to CMT recipients will aid in the development of an effective CMV vaccine both as therapeutic and prophylactic agent.
| Project Number : | 5P01CA030206-230013 |
|---|
| ICD : | NATIONAL CANCER INSTITUTE |
|---|
| Project Terms : | cytomegalovirus, human therapy evaluation, immunity, vaccine development, viral vaccine Herpesviridae disease, bone marrow transplantation, cytotoxic T lymphocyte, homologous transplantation, immunologic memory, microorganism immunology, nonhuman therapy evaluation, virus protein clinical research, epitope mapping, flow cytometry, genetically modified animal, human subject, laboratory mouse, peptide library |
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VACCINATION FOR CMV IN AIDS PATIENTS UNDERGOING HAART
(2003)
Abstract :
DESCRIPTION (Adapted from the Investigator's Abstract): Based on cytotoxic T lymphocyte (CTL) epitopes from CMV pp65, the principal CTL target for CMV, a novel candidate lipopeptide vaccine has been developed. It is composed of the HLA A*0201-restricted CTL epitope from pp65 covalently attached to a powerful synthetic T-helper epitope, and lipidated at the amino terminus. The test-of-concept for this novel lipopeptide CMV vaccine has been shown using an HLA A2.1 transgenic murine model. The goal of this project is to characterize the immune response to CMV in persons treated with HAART (highly active anti-retroviral therapy) and determine if this immunity can be boosted by lipopeptide immunization. In a three-way collaboration, the City of Hope team will conduct the in vitro measurements of immunity; and the team at the Los Angeles County Medical Center/University of Southern California (LAC/USC) will identify patients, obtain specimens for in vitro measurements, and vaccinate eligible patients. The third team, Peninsula Laboratories, will be responsible for the synthesis and production of lipopeptide vaccine under FDA-approved conditions. Individuals with HIV infection will be studied to understand the cellular immune response to CMV. Information not currently known despite the fact that CMV-associated diseases, including retinitis, enteritis, and encephalitis have been a leading cause of morbidity and mortality in this population, and that CMV could be linked to HIV progression. These studies are important because of the need to explain recent reports indicating less CMV disease with HAART, but the occasional case of CMV retinitis despite recovery to >200 CD4+ T cell counts. At LAC/USC, approximately 70% of new AIDS clinic patients are naive to HAART and will be evaluated for changing CMV immune repertoire during the course of HAART treatment and CD4+ T cell recovery. These studies are aimed at discovering whether immunoprophylaxis against CMV is possible or warranted in individuals undergoing HAART. The strategy of immunization in the setting of HIV-1 infection and HAART in patients with diverse DLA alleles will be built upon a concurrent demonstration of the clinical efficacy of the HLA A2.1-restricted CMV pp65 lipopeptide vaccine in patient volunteers.
| Project Number : | 3R01AI043267-04S1 |
|---|
| ICD : | NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES |
|---|
| IRG : | ZRG5 |
|---|
| Project Terms : | AIDS, AIDS therapy, Herpesviridae disease, Herpesviridae vaccine, active immunization, cytomegalovirus, human therapy evaluation cellular immunity, cytomegalovirus retinitis, cytotoxic T lymphocyte, drug design /synthesis /production, helper T lymphocyte, human immunodeficiency virus 1, vaccine development clinical research, human subject, tissue /cell culture |
|---|
VACCINATION FOR CMV IN AIDS PATIENTS UNDERGOING HAART
(2003)
Abstract :
DESCRIPTION (Adapted from the Investigator's Abstract): Based on cytotoxic T lymphocyte (CTL) epitopes from CMV pp65, the principal CTL target for CMV, a novel candidate lipopeptide vaccine has been developed. It is composed of the HLA A*0201-restricted CTL epitope from pp65 covalently attached to a powerful synthetic T-helper epitope, and lipidated at the amino terminus. The test-of-concept for this novel lipopeptide CMV vaccine has been shown using an HLA A2.1 transgenic murine model. The goal of this project is to characterize the immune response to CMV in persons treated with HAART (highly active anti-retroviral therapy) and determine if this immunity can be boosted by lipopeptide immunization. In a three-way collaboration, the City of Hope team will conduct the in vitro measurements of immunity; and the team at the Los Angeles County Medical Center/University of Southern California (LAC/USC) will identify patients, obtain specimens for in vitro measurements, and vaccinate eligible patients. The third team, Peninsula Laboratories, will be responsible for the synthesis and production of lipopeptide vaccine under FDA-approved conditions. Individuals with HIV infection will be studied to understand the cellular immune response to CMV. Information not currently known despite the fact that CMV-associated diseases, including retinitis, enteritis, and encephalitis have been a leading cause of morbidity and mortality in this population, and that CMV could be linked to HIV progression. These studies are important because of the need to explain recent reports indicating less CMV disease with HAART, but the occasional case of CMV retinitis despite recovery to >200 CD4+ T cell counts. At LAC/USC, approximately 70% of new AIDS clinic patients are naive to HAART and will be evaluated for changing CMV immune repertoire during the course of HAART treatment and CD4+ T cell recovery. These studies are aimed at discovering whether immunoprophylaxis against CMV is possible or warranted in individuals undergoing HAART. The strategy of immunization in the setting of HIV-1 infection and HAART in patients with diverse DLA alleles will be built upon a concurrent demonstration of the clinical efficacy of the HLA A2.1-restricted CMV pp65 lipopeptide vaccine in patient volunteers.
| Project Number : | 5R01AI043267-04 |
|---|
| ICD : | NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES |
|---|
| IRG : | ZRG5 |
|---|
| Project Terms : | AIDS, AIDS therapy, Herpesviridae disease, Herpesviridae vaccine, active immunization, cytomegalovirus, human therapy evaluation cellular immunity, cytomegalovirus retinitis, cytotoxic T lymphocyte, drug design /synthesis /production, helper T lymphocyte, human immunodeficiency virus 1, vaccine development clinical research, human subject, tissue /cell culture |
|---|
LIPOPEPTIDE VACCINE TO PREVENT HCMV INFECTION AFTER BMT
(2003)
Abstract :
DESCRIPTION: (Applicant's Abstract) A complication of the recovery phase after BMT is cytomegalovirus (HCMV) infection. Therapy using gancyclovir prophylaxis has decreased the mortality from HCMV infection in allogeneic BMT recipients. However side-effects which include delayed immune reconstitution limit its effectiveness in reducing overall mortality from BMT. Studies from several laboratories have indicated that control of CMV infection in BMT recipients is dependent on a strong cytotoxic T lymphocyte (CTL) response which is targeted mainly against the viral matrix protein pp65, and to a lesser degree against pp150. An adoptive immunotherapy strategy using CTL has proved successful but is impractical for general use in BMT centers. The applicant will develop an alternative strategy that uses small peptide fragments from the CTL viral target proteins pp65 and pp150 in the form of a vaccine as a means to simulate CTL in vivo. The vaccine is a peptide composed of a CTL epitope combined with a peptide which stimulates a T helper (TH) response and is lipidated on the amino terminus. He will first evaluate several different structural forms of the vaccine containing the HLA-A0201 restricted CTL epitope from pp65 and several different TH epitopes in a transgenic HLA-A2.1 mouse model. He will evaluate which of the vaccine molecules provide the strongest and longest-lived immunity by comparing the degree of lysis in vitro of human fibroblasts infected with HCMV mediated by murine splenic effector CTL. Based on the results of murine immunizations, a Phase I dose-escalating and safety trial will be conducted in asymptomatic HCMV seropositive human volunteers with a lipidated HLA-A0201 restricted CTL vaccine. Additional vaccines including those with 2 or 3 CTL epitopes will also be evaluated in human volunteers in a Phase I trial. In order to test whether the vaccine, which enhances CTL against HCMV in healthy volunteers, is also capable of protecting against infection, a Phase II trial will be conducted in HCMV seropositive BMT donors and recipients. BMT donors will be vaccinated according to a protocol determined by the Phase I trial and their HLA-matched siblings who undergo BMT will be monitored for the ability to resist viremia and/or pneumonia brought about by HCMV infection. Success of this protocol will be determined by the avoidance of gancyclovir post-BMT and reduction of early and late HCMV infection in the BMT recipient.
| Project Number : | 5R01CA077544-04 |
|---|
| ICD : | NATIONAL CANCER INSTITUTE |
|---|
| IRG : | ET |
|---|
| Project Terms : | bone marrow transplantation, chemoprevention, cytomegalovirus, drug screening /evaluation, postoperative complication, synthetic vaccine, viral vaccine, virus infection mechanism MHC class I antigen, active immunization, clinical trial phase I, clinical trial phase II /III /IV, cytotoxic T lymphocyte, dosage, drug adverse effect, extracellular matrix protein, helper T lymphocyte, viremia, virus protein clinical research, human subject, laboratory mouse, lung lavage, polymerase chain reaction, transgenic animal |
|---|
LIPOPEPTIDE VACCINE TO PREVENT HCMV INFECTION AFTER BMT
(2003)
Abstract :
DESCRIPTION: (Applicant's Abstract) A complication of the recovery phase after BMT is cytomegalovirus (HCMV) infection. Therapy using gancyclovir prophylaxis has decreased the mortality from HCMV infection in allogeneic BMT recipients. However side-effects which include delayed immune reconstitution limit its effectiveness in reducing overall mortality from BMT. Studies from several laboratories have indicated that control of CMV infection in BMT recipients is dependent on a strong cytotoxic T lymphocyte (CTL) response which is targeted mainly against the viral matrix protein pp65, and to a lesser degree against pp150. An adoptive immunotherapy strategy using CTL has proved successful but is impractical for general use in BMT centers. The applicant will develop an alternative strategy that uses small peptide fragments from the CTL viral target proteins pp65 and pp150 in the form of a vaccine as a means to simulate CTL in vivo. The vaccine is a peptide composed of a CTL epitope combined with a peptide which stimulates a T helper (TH) response and is lipidated on the amino terminus. He will first evaluate several different structural forms of the vaccine containing the HLA-A0201 restricted CTL epitope from pp65 and several different TH epitopes in a transgenic HLA-A2.1 mouse model. He will evaluate which of the vaccine molecules provide the strongest and longest-lived immunity by comparing the degree of lysis in vitro of human fibroblasts infected with HCMV mediated by murine splenic effector CTL. Based on the results of murine immunizations, a Phase I dose-escalating and safety trial will be conducted in asymptomatic HCMV seropositive human volunteers with a lipidated HLA-A0201 restricted CTL vaccine. Additional vaccines including those with 2 or 3 CTL epitopes will also be evaluated in human volunteers in a Phase I trial. In order to test whether the vaccine, which enhances CTL against HCMV in healthy volunteers, is also capable of protecting against infection, a Phase II trial will be conducted in HCMV seropositive BMT donors and recipients. BMT donors will be vaccinated according to a protocol determined by the Phase I trial and their HLA-matched siblings who undergo BMT will be monitored for the ability to resist viremia and/or pneumonia brought about by HCMV infection. Success of this protocol will be determined by the avoidance of gancyclovir post-BMT and reduction of early and late HCMV infection in the BMT recipient.
| Project Number : | 1R01CA077544-01 |
|---|
| ICD : | NATIONAL CANCER INSTITUTE |
|---|
| IRG : | ET |
|---|
| Project Terms : | bone marrow transplantation, chemoprevention, cytomegalovirus, drug screening /evaluation, postoperative complication, synthetic vaccine, viral vaccine, virus infection mechanism MHC class I antigen, active immunization, clinical trial phase I, clinical trial phase II /III /IV, cytotoxic T lymphocyte, dosage, drug adverse effect, extracellular matrix protein, helper T lymphocyte, viremia, virus protein clinical research, human subject, laboratory mouse, lung lavage, polymerase chain reaction, transgenic animal |
|---|
LIPOPEPTIDE VACCINE TO PREVENT HCMV INFECTION AFTER BMT
(2003)
Abstract :
DESCRIPTION: (Applicant's Abstract) A complication of the recovery phase after BMT is cytomegalovirus (HCMV) infection. Therapy using gancyclovir prophylaxis has decreased the mortality from HCMV infection in allogeneic BMT recipients. However side-effects which include delayed immune reconstitution limit its effectiveness in reducing overall mortality from BMT. Studies from several laboratories have indicated that control of CMV infection in BMT recipients is dependent on a strong cytotoxic T lymphocyte (CTL) response which is targeted mainly against the viral matrix protein pp65, and to a lesser degree against pp150. An adoptive immunotherapy strategy using CTL has proved successful but is impractical for general use in BMT centers. The applicant will develop an alternative strategy that uses small peptide fragments from the CTL viral target proteins pp65 and pp150 in the form of a vaccine as a means to simulate CTL in vivo. The vaccine is a peptide composed of a CTL epitope combined with a peptide which stimulates a T helper (TH) response and is lipidated on the amino terminus. He will first evaluate several different structural forms of the vaccine containing the HLA-A0201 restricted CTL epitope from pp65 and several different TH epitopes in a transgenic HLA-A2.1 mouse model. He will evaluate which of the vaccine molecules provide the strongest and longest-lived immunity by comparing the degree of lysis in vitro of human fibroblasts infected with HCMV mediated by murine splenic effector CTL. Based on the results of murine immunizations, a Phase I dose-escalating and safety trial will be conducted in asymptomatic HCMV seropositive human volunteers with a lipidated HLA-A0201 restricted CTL vaccine. Additional vaccines including those with 2 or 3 CTL epitopes will also be evaluated in human volunteers in a Phase I trial. In order to test whether the vaccine, which enhances CTL against HCMV in healthy volunteers, is also capable of protecting against infection, a Phase II trial will be conducted in HCMV seropositive BMT donors and recipients. BMT donors will be vaccinated according to a protocol determined by the Phase I trial and their HLA-matched siblings who undergo BMT will be monitored for the ability to resist viremia and/or pneumonia brought about by HCMV infection. Success of this protocol will be determined by the avoidance of gancyclovir post-BMT and reduction of early and late HCMV infection in the BMT recipient.
| Project Number : | 5R01CA077544-02 |
|---|
| ICD : | NATIONAL CANCER INSTITUTE |
|---|
| IRG : | ET |
|---|
| Project Terms : | bone marrow transplantation, chemoprevention, cytomegalovirus, drug screening /evaluation, postoperative complication, synthetic vaccine, viral vaccine, virus infection mechanism MHC class I antigen, active immunization, clinical trial phase I, clinical trial phase II /III /IV, cytotoxic T lymphocyte, dosage, drug adverse effect, extracellular matrix protein, helper T lymphocyte, viremia, virus protein clinical research, human subject, laboratory mouse, lung lavage, polymerase chain reaction, transgenic animal |
|---|
LIPOPEPTIDE VACCINE TO PREVENT HCMV INFECTION AFTER BMT
(2003)
Abstract :
DESCRIPTION: (Applicant's Abstract) A complication of the recovery phase after BMT is cytomegalovirus (HCMV) infection. Therapy using gancyclovir prophylaxis has decreased the mortality from HCMV infection in allogeneic BMT recipients. However side-effects which include delayed immune reconstitution limit its effectiveness in reducing overall mortality from BMT. Studies from several laboratories have indicated that control of CMV infection in BMT recipients is dependent on a strong cytotoxic T lymphocyte (CTL) response which is targeted mainly against the viral matrix protein pp65, and to a lesser degree against pp150. An adoptive immunotherapy strategy using CTL has proved successful but is impractical for general use in BMT centers. The applicant will develop an alternative strategy that uses small peptide fragments from the CTL viral target proteins pp65 and pp150 in the form of a vaccine as a means to simulate CTL in vivo. The vaccine is a peptide composed of a CTL epitope combined with a peptide which stimulates a T helper (TH) response and is lipidated on the amino terminus. He will first evaluate several different structural forms of the vaccine containing the HLA-A0201 restricted CTL epitope from pp65 and several different TH epitopes in a transgenic HLA-A2.1 mouse model. He will evaluate which of the vaccine molecules provide the strongest and longest-lived immunity by comparing the degree of lysis in vitro of human fibroblasts infected with HCMV mediated by murine splenic effector CTL. Based on the results of murine immunizations, a Phase I dose-escalating and safety trial will be conducted in asymptomatic HCMV seropositive human volunteers with a lipidated HLA-A0201 restricted CTL vaccine. Additional vaccines including those with 2 or 3 CTL epitopes will also be evaluated in human volunteers in a Phase I trial. In order to test whether the vaccine, which enhances CTL against HCMV in healthy volunteers, is also capable of protecting against infection, a Phase II trial will be conducted in HCMV seropositive BMT donors and recipients. BMT donors will be vaccinated according to a protocol determined by the Phase I trial and their HLA-matched siblings who undergo BMT will be monitored for the ability to resist viremia and/or pneumonia brought about by HCMV infection. Success of this protocol will be determined by the avoidance of gancyclovir post-BMT and reduction of early and late HCMV infection in the BMT recipient.
| Project Number : | 5R01CA077544-03 |
|---|
| ICD : | NATIONAL CANCER INSTITUTE |
|---|
| IRG : | ET |
|---|
| Project Terms : | bone marrow transplantation, chemoprevention, cytomegalovirus, drug screening /evaluation, postoperative complication, synthetic vaccine, viral vaccine, virus infection mechanism MHC class I antigen, active immunization, clinical trial phase I, clinical trial phase II /III /IV, cytotoxic T lymphocyte, dosage, drug adverse effect, extracellular matrix protein, helper T lymphocyte, viremia, virus protein clinical research, human subject, laboratory mouse, lung lavage, polymerase chain reaction, transgenic animal |
|---|
ACQUISITION OF IMMUNITY TO CYTOMEGALOVIRUS
(2003)
Abstract :
CMV infection remains an important problem for transplantation. No successful vaccine has been developed that prevents new infections or even controls existing ones. Pharmacologic treatments have limitations regarding their side effects and the development of resistant strain Asymptomatic CMV-seropositive individuals of survivors of CMV infection after BMT, have an ongoing cytotoxic T lymphocyte (CTL) response to CMV, which can be measured after in vitro stimulation (IVS), either using HLA tetrameter binding or chromium release assay (CRA). The CTL response has multiple targets, however, the response to the tegument protein pp65 predominates in most individuals examined. Although many CTL epitopes have already been mapped from two immunodominant CMV proteins (pp65 and pp150), a significant number have yet been determined, and their elucidation will lead to a broad-based vaccine strategy for all major ethic groups in the United States. Immunodominance of CTL responses to CMV proteins will be determined utilizing IVS strategies together with known CTL epitopes. In contrast, little is known of the predominance of CTL targets from CMV in BMT recipients. The frequency of CMV-specific CTL will be enumerated using flow cytometry with HLA tetramers, and the frequency of those CTL will be compared to viral load measurements using plasma PCR methods. As a means to enhance the immunogenicity of previous defined CTL epitopes, a combinatorial peptide chemistry approach will be used with the known CTL epitopes from pp65 and pp150, which have affinities that are in the micromolar to approach will be used with the known CTL epitopes from pp65 and pp150, which have affinities that are in the micromolar to nanomolar range. Analogue peptides will be defined that will be evaluated initially using HLA A*0201 restricted and pp65- specific T cell clones, to develop a more immunogenic CTL epitope which may lead to a more effective CMV vaccine. The functionality of these analogue peptides will be shown utilizing in vivo immunization of HLA-transgenic mice and IVS studies stimulating a memory response in human PBMC specific for killing of CMV infected fibroblasts in vitro. Two Phase II trials will evaluate modalities including CMV lipopeptide immunization in combination with standard tetanus toxin immunization of BMT recipients. Augmentation of the memory response to tetanus- specific CD4+ T cells will be evaluated by in vitro methods, and the duration of infection-free survival of BMT recipients will be measured using standard clinical parameters. Finally, the utility of targeting a single CMV protein utilizing a CTL epitope vaccine strategy versus targeting two or more CMV proteins will be evaluated using CMV lipopeptide immunization of BMT recipients. Since the endpoint of the trial is a reduction in the incidence of CMV-associated disease, the comparison must be done utilizing a patient population likely to have measurable CMV viral load. Taken together, these strategies of defining CTL epitopes, augmenting their immunogenicity, and defining efficacious ways to delay them to CMT recipients will aid in the development of an effective CMV vaccine both as therapeutic and prophylactic agent.
| Project Number : | 6P01CA030206-220013 |
|---|
| ICD : | NATIONAL CANCER INSTITUTE |
|---|
| Project Terms : | cytomegalovirus, human therapy evaluation, immunity, vaccine development, viral vaccine Herpesviridae disease, bone marrow transplantation, clinical trial phase II, cytotoxic T lymphocyte, homologous transplantation, immunologic memory, microorganism immunology, nonhuman therapy evaluation, virus protein clinical research, epitope mapping, flow cytometry, human subject, laboratory mouse, patient oriented research, peptide library, transgenic animal |
|---|
LIPOPEPTIDE VACCINE AGAINST CONSERVED TARGETS OF HIV
(2002)
Abstract :
DESCRIPTION: (Adapted from applicant's abstract): The cellular immune response to HIV has been shown to be robust and continues throughout the natural history of the infection. The capacity for CTL to diminish viral load is substantial, however, sequence variation of HIV limits the effectiveness of the CTL response. Combination pharmacologic therapy against HIV infection known as HAART, has dramatically decreased viral load in many AIDS patients. Despite this control of viral replication, patients who are HAART-responsive for a period of two years or greater, still harbor infectious virus that may, under the appropriate circumstance, become active in replication and cause serious health consequences. The purpose of these studies is to develop immunization strategies in murine models based on immunodominant CTL epitopes from HIV proteins in combination with T-help (HTL) epitopes as part of lipidated peptides. Epitopes will be chosen from sequence-conserved regions of immunogenic HIV proteins. These epitopes will be made into combination vaccines consisting of covalently attached and lipidated HTL+CTL epitopes. The initial approach is to determine the most efficacious means using lipidated vaccines to immunize transgenic mice expressing human HLA Class I and Class II MHC antigens. Mucosal and subcutaneous routes of immunization will be explored. The success of immunization strategies will be evaluated by in vitro methods including the recognition of human antigen presenting cells (APC) which will be infected with different isolates of HIV. The choice of CTL epitope will be made using the criteria that the recognition should be restricted by common HLA Class I alleles. One of the main goals of the proposal and the Innovation Grant Program is to evaluate means to propagate the CTL response using strong T-help (HTL) epitopes. The applicants will evaluate a series of HTL epitopes in combination with selected CTL epitopes using immunization strategies detailed in the proposal. The efficacy of the immune strategies will be examined further using a mouse model of HIV infection referred to as the huPBL-SCID mouse. Immunodeficient mice will be repopulated with human PBL of known HLA type and infected with various isolates of HIV. Vaccine stimulated CTL obtained from transgenic mice will be infused in the previously HIV infected huPBL SCID mice to evaluate their potential for recognition/lysis of infected cells and reduction of viral load. The vaccine strategy that results in the most successful reduction in viral load should be considered in terms of future human immunization clinical protocols for patients on HAART during the time when viral replication is suppressed.
| Project Number : | 5R21AI044313-02 |
|---|
| ICD : | NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES |
|---|
| IRG : | ZAI1 |
|---|
| Project Terms : | AIDS vaccine, HIV infection, cellular immunity, cytotoxic T lymphocyte, nonblood lipoprotein, peptide, vaccine development MHC class I antigen, antigen presenting cell, disease model, drug screening /evaluation, helper T lymphocyte, human immunodeficiency virus, passive immunization, protein sequence, viremia, virus protein, virus replication NOD mouse, SCID mouse, cell line, clinical research, drug administration route, human subject, transgenic animal |
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VACCINATION FOR CMV IN AIDS PATIENTS UNDERGOING HAART
(2002)
Abstract :
DESCRIPTION (Adapted from the Investigator's Abstract): Based on cytotoxic T lymphocyte (CTL) epitopes from CMV pp65, the principal CTL target for CMV, a novel candidate lipopeptide vaccine has been developed. It is composed of the HLA A*0201-restricted CTL epitope from pp65 covalently attached to a powerful synthetic T-helper epitope, and lipidated at the amino terminus. The test-of-concept for this novel lipopeptide CMV vaccine has been shown using an HLA A2.1 transgenic murine model. The goal of this project is to characterize the immune response to CMV in persons treated with HAART (highly active anti-retroviral therapy) and determine if this immunity can be boosted by lipopeptide immunization. In a three-way collaboration, the City of Hope team will conduct the in vitro measurements of immunity; and the team at the Los Angeles County Medical Center/University of Southern California (LAC/USC) will identify patients, obtain specimens for in vitro measurements, and vaccinate eligible patients. The third team, Peninsula Laboratories, will be responsible for the synthesis and production of lipopeptide vaccine under FDA-approved conditions. Individuals with HIV infection will be studied to understand the cellular immune response to CMV. Information not currently known despite the fact that CMV-associated diseases, including retinitis, enteritis, and encephalitis have been a leading cause of morbidity and mortality in this population, and that CMV could be linked to HIV progression. These studies are important because of the need to explain recent reports indicating less CMV disease with HAART, but the occasional case of CMV retinitis despite recovery to >200 CD4+ T cell counts. At LAC/USC, approximately 70% of new AIDS clinic patients are naive to HAART and will be evaluated for changing CMV immune repertoire during the course of HAART treatment and CD4+ T cell recovery. These studies are aimed at discovering whether immunoprophylaxis against CMV is possible or warranted in individuals undergoing HAART. The strategy of immunization in the setting of HIV-1 infection and HAART in patients with diverse DLA alleles will be built upon a concurrent demonstration of the clinical efficacy of the HLA A2.1-restricted CMV pp65 lipopeptide vaccine in patient volunteers.
| Project Number : | 5R01AI043267-02 |
|---|
| ICD : | NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES |
|---|
| IRG : | ZRG5 |
|---|
| Project Terms : | AIDS, AIDS therapy, Herpesviridae disease, Herpesviridae vaccine, active immunization, cytomegalovirus, human therapy evaluation cellular immunity, cytomegalovirus retinitis, cytotoxic T lymphocyte, drug design /synthesis /production, helper T lymphocyte, human immunodeficiency virus 1, vaccine development clinical research, human subject, tissue /cell culture |
|---|
VACCINATION FOR CMV IN AIDS PATIENTS UNDERGOING HAART
(2002)
Abstract :
DESCRIPTION (Adapted from the Investigator's Abstract): Based on cytotoxic T lymphocyte (CTL) epitopes from CMV pp65, the principal CTL target for CMV, a novel candidate lipopeptide vaccine has been developed. It is composed of the HLA A*0201-restricted CTL epitope from pp65 covalently attached to a powerful synthetic T-helper epitope, and lipidated at the amino terminus. The test-of-concept for this novel lipopeptide CMV vaccine has been shown using an HLA A2.1 transgenic murine model. The goal of this project is to characterize the immune response to CMV in persons treated with HAART (highly active anti-retroviral therapy) and determine if this immunity can be boosted by lipopeptide immunization. In a three-way collaboration, the City of Hope team will conduct the in vitro measurements of immunity; and the team at the Los Angeles County Medical Center/University of Southern California (LAC/USC) will identify patients, obtain specimens for in vitro measurements, and vaccinate eligible patients. The third team, Peninsula Laboratories, will be responsible for the synthesis and production of lipopeptide vaccine under FDA-approved conditions. Individuals with HIV infection will be studied to understand the cellular immune response to CMV. Information not currently known despite the fact that CMV-associated diseases, including retinitis, enteritis, and encephalitis have been a leading cause of morbidity and mortality in this population, and that CMV could be linked to HIV progression. These studies are important because of the need to explain recent reports indicating less CMV disease with HAART, but the occasional case of CMV retinitis despite recovery to >200 CD4+ T cell counts. At LAC/USC, approximately 70% of new AIDS clinic patients are naive to HAART and will be evaluated for changing CMV immune repertoire during the course of HAART treatment and CD4+ T cell recovery. These studies are aimed at discovering whether immunoprophylaxis against CMV is possible or warranted in individuals undergoing HAART. The strategy of immunization in the setting of HIV-1 infection and HAART in patients with diverse DLA alleles will be built upon a concurrent demonstration of the clinical efficacy of the HLA A2.1-restricted CMV pp65 lipopeptide vaccine in patient volunteers.
| Project Number : | 5R01AI043267-03 |
|---|
| ICD : | NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES |
|---|
| IRG : | ZRG5 |
|---|
| Project Terms : | AIDS, AIDS therapy, Herpesviridae disease, Herpesviridae vaccine, active immunization, cytomegalovirus, human therapy evaluation cellular immunity, cytomegalovirus retinitis, cytotoxic T lymphocyte, drug design /synthesis /production, helper T lymphocyte, human immunodeficiency virus 1, vaccine development clinical research, human subject, tissue /cell culture |
|---|
VACCINATION FOR CMV IN AIDS PATIENTS UNDERGOING HAART
(2002)
Abstract :
DESCRIPTION (Adapted from the Investigator's Abstract): Based on cytotoxic T lymphocyte (CTL) epitopes from CMV pp65, the principal CTL target for CMV, a novel candidate lipopeptide vaccine has been developed. It is composed of the HLA A*0201-restricted CTL epitope from pp65 covalently attached to a powerful synthetic T-helper epitope, and lipidated at the amino terminus. The test-of-concept for this novel lipopeptide CMV vaccine has been shown using an HLA A2.1 transgenic murine model. The goal of this project is to characterize the immune response to CMV in persons treated with HAART (highly active anti-retroviral therapy) and determine if this immunity can be boosted by lipopeptide immunization. In a three-way collaboration, the City of Hope team will conduct the in vitro measurements of immunity; and the team at the Los Angeles County Medical Center/University of Southern California (LAC/USC) will identify patients, obtain specimens for in vitro measurements, and vaccinate eligible patients. The third team, Peninsula Laboratories, will be responsible for the synthesis and production of lipopeptide vaccine under FDA-approved conditions. Individuals with HIV infection will be studied to understand the cellular immune response to CMV. Information not currently known despite the fact that CMV-associated diseases, including retinitis, enteritis, and encephalitis have been a leading cause of morbidity and mortality in this population, and that CMV could be linked to HIV progression. These studies are important because of the need to explain recent reports indicating less CMV disease with HAART, but the occasional case of CMV retinitis despite recovery to >200 CD4+ T cell counts. At LAC/USC, approximately 70% of new AIDS clinic patients are naive to HAART and will be evaluated for changing CMV immune repertoire during the course of HAART treatment and CD4+ T cell recovery. These studies are aimed at discovering whether immunoprophylaxis against CMV is possible or warranted in individuals undergoing HAART. The strategy of immunization in the setting of HIV-1 infection and HAART in patients with diverse DLA alleles will be built upon a concurrent demonstration of the clinical efficacy of the HLA A2.1-restricted CMV pp65 lipopeptide vaccine in patient volunteers.
| Project Number : | 3R01AI043267-02S1 |
|---|
| ICD : | NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES |
|---|
| IRG : | ZRG5 |
|---|
| Project Terms : | AIDS, AIDS therapy, Herpesviridae disease, Herpesviridae vaccine, active immunization, cytomegalovirus, human therapy evaluation cellular immunity, cytomegalovirus retinitis, cytotoxic T lymphocyte, drug design /synthesis /production, helper T lymphocyte, human immunodeficiency virus 1, vaccine development clinical research, human subject, tissue /cell culture |
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LIPOPEPTIDE VACCINE W/ ACTIVITY AGAINST HUMAN CYTOMEGALOVIRUS
(2002)
| Project Number : | 5M01RR000043-420736 |
|---|
| ICD : | NATIONAL CENTER FOR RESEARCH RESOURCES |
|---|
| Project Terms : | cytomegalovirus, dosage, drug screening /evaluation, vaccine development, viral vaccine clinical trial phase I, immunologic memory adult human (21+), clinical research, human subject |
|---|
BONE MARROW DONORS & VOLUNTEERS FOR STUDY OF CMV
(2002)
| Project Number : | 5M01RR000043-420738 |
|---|
| ICD : | NATIONAL CENTER FOR RESEARCH RESOURCES |
|---|
| Project Terms : | bone marrow transplantation, cytomegalovirus, tissue donor, vaccine development, viral vaccine, virus antigen microorganism immunology clinical research, human subject |
|---|
ACQUISITION OF IMMUNITY TO CYTOMEGALOVIRUS
(2002)
Abstract :
CMV infection remains an important problem for transplantation. No successful vaccine has been developed that prevents new infections or even controls existing ones. Pharmacologic treatments have limitations regarding their side effects and the development of resistant strain Asymptomatic CMV-seropositive individuals of survivors of CMV infection after BMT, have an ongoing cytotoxic T lymphocyte (CTL) response to CMV, which can be measured after in vitro stimulation (IVS), either using HLA tetrameter binding or chromium release assay (CRA). The CTL response has multiple targets, however, the response to the tegument protein pp65 predominates in most individuals examined. Although many CTL epitopes have already been mapped from two immunodominant CMV proteins (pp65 and pp150), a significant number have yet been determined, and their elucidation will lead to a broad-based vaccine strategy for all major ethic groups in the United States. Immunodominance of CTL responses to CMV proteins will be determined utilizing IVS strategies together with known CTL epitopes. In contrast, little is known of the predominance of CTL targets from CMV in BMT recipients. The frequency of CMV-specific CTL will be enumerated using flow cytometry with HLA tetramers, and the frequency of those CTL will be compared to viral load measurements using plasma PCR methods. As a means to enhance the immunogenicity of previous defined CTL epitopes, a combinatorial peptide chemistry approach will be used with the known CTL epitopes from pp65 and pp150, which have affinities that are in the micromolar to approach will be used with the known CTL epitopes from pp65 and pp150, which have affinities that are in the micromolar to nanomolar range. Analogue peptides will be defined that will be evaluated initially using HLA A*0201 restricted and pp65- specific T cell clones, to develop a more immunogenic CTL epitope which may lead to a more effective CMV vaccine. The functionality of these analogue peptides will be shown utilizing in vivo immunization of HLA-transgenic mice and IVS studies stimulating a memory response in human PBMC specific for killing of CMV infected fibroblasts in vitro. Two Phase II trials will evaluate modalities including CMV lipopeptide immunization in combination with standard tetanus toxin immunization of BMT recipients. Augmentation of the memory response to tetanus- specific CD4+ T cells will be evaluated by in vitro methods, and the duration of infection-free survival of BMT recipients will be measured using standard clinical parameters. Finally, the utility of targeting a single CMV protein utilizing a CTL epitope vaccine strategy versus targeting two or more CMV proteins will be evaluated using CMV lipopeptide immunization of BMT recipients. Since the endpoint of the trial is a reduction in the incidence of CMV-associated disease, the comparison must be done utilizing a patient population likely to have measurable CMV viral load. Taken together, these strategies of defining CTL epitopes, augmenting their immunogenicity, and defining efficacious ways to delay them to CMT recipients will aid in the development of an effective CMV vaccine both as therapeutic and prophylactic agent.
| Project Number : | 5P01CA030206-210013 |
|---|
| ICD : | NATIONAL CANCER INSTITUTE |
|---|
| Project Terms : | cytomegalovirus, human therapy evaluation, immunity, vaccine development, viral vaccine Herpesviridae disease, bone marrow transplantation, clinical trial phase II /III /IV, cytotoxic T lymphocyte, homologous transplantation, immunologic memory, microorganism immunology, nonhuman therapy evaluation, virus protein clinical research, epitope mapping, flow cytometry, human subject, laboratory mouse, peptide library, transgenic animal |
|---|
ACQUISITION OF IMMUNITY TO CYTOMEGALOVIRUS
(2001)
Abstract :
CMV infection remains an important problem for transplantation. No successful vaccine has been developed that prevents new infections or even controls existing ones. Pharmacologic treatments have limitations regarding their side effects and the development of resistant strain Asymptomatic CMV-seropositive individuals of survivors of CMV infection after BMT, have an ongoing cytotoxic T lymphocyte (CTL) response to CMV, which can be measured after in vitro stimulation (IVS), either using HLA tetrameter binding or chromium release assay (CRA). The CTL response has multiple targets, however, the response to the tegument protein pp65 predominates in most individuals examined. Although many CTL epitopes have already been mapped from two immunodominant CMV proteins (pp65 and pp150), a significant number have yet been determined, and their elucidation will lead to a broad-based vaccine strategy for all major ethic groups in the United States. Immunodominance of CTL responses to CMV proteins will be determined utilizing IVS strategies together with known CTL epitopes. In contrast, little is known of the predominance of CTL targets from CMV in BMT recipients. The frequency of CMV-specific CTL will be enumerated using flow cytometry with HLA tetramers, and the frequency of those CTL will be compared to viral load measurements using plasma PCR methods. As a means to enhance the immunogenicity of previous defined CTL epitopes, a combinatorial peptide chemistry approach will be used with the known CTL epitopes from pp65 and pp150, which have affinities that are in the micromolar to approach will be used with the known CTL epitopes from pp65 and pp150, which have affinities that are in the micromolar to nanomolar range. Analogue peptides will be defined that will be evaluated initially using HLA A*0201 restricted and pp65- specific T cell clones, to develop a more immunogenic CTL epitope which may lead to a more effective CMV vaccine. The functionality of these analogue peptides will be shown utilizing in vivo immunization of HLA-transgenic mice and IVS studies stimulating a memory response in human PBMC specific for killing of CMV infected fibroblasts in vitro. Two Phase II trials will evaluate modalities including CMV lipopeptide immunization in combination with standard tetanus toxin immunization of BMT recipients. Augmentation of the memory response to tetanus- specific CD4+ T cells will be evaluated by in vitro methods, and the duration of infection-free survival of BMT recipients will be measured using standard clinical parameters. Finally, the utility of targeting a single CMV protein utilizing a CTL epitope vaccine strategy versus targeting two or more CMV proteins will be evaluated using CMV lipopeptide immunization of BMT recipients. Since the endpoint of the trial is a reduction in the incidence of CMV-associated disease, the comparison must be done utilizing a patient population likely to have measurable CMV viral load. Taken together, these strategies of defining CTL epitopes, augmenting their immunogenicity, and defining efficacious ways to delay them to CMT recipients will aid in the development of an effective CMV vaccine both as therapeutic and prophylactic agent.
| Project Number : | 5P01CA030206-200013 |
|---|
| ICD : | NATIONAL CANCER INSTITUTE |
|---|
| Project Terms : | cytomegalovirus, human therapy evaluation, immunity, vaccine development, viral vaccine Herpesviridae disease, bone marrow transplantation, clinical trial phase II /III /IV, cytotoxic T lymphocyte, homologous transplantation, immunologic memory, microorganism immunology, nonhuman therapy evaluation, virus protein clinical research, epitope mapping, flow cytometry, human subject, laboratory mouse, peptide library, transgenic animal |
|---|
LIPOPEPTIDE VACCINE W/ ACTIVITY AGAINST HUMAN CYTOMEGALOVIRUS
(2001)
| Project Number : | 3M01RR000043-41S10736 |
|---|
| ICD : | NATIONAL CENTER FOR RESEARCH RESOURCES |
|---|
| Project Terms : | cytomegalovirus, dosage, drug screening /evaluation, vaccine development, viral vaccine clinical trial phase I, immunologic memory adult human (19+), clinical research, human subject |
|---|
BONE MARROW DONORS & VOLUNTEERS FOR STUDY OF CMV
(2001)
| Project Number : | 3M01RR000043-41S10738 |
|---|
| ICD : | NATIONAL CENTER FOR RESEARCH RESOURCES |
|---|
| Project Terms : | bone marrow transplantation, cytomegalovirus, tissue donor, vaccine development, viral vaccine, virus antigen microorganism immunology clinical research, human subject |
|---|
LIPOPEPTIDE VACCINE W/ ACTIVITY AGAINST HUMAN CYTOMEGALOVIRUS
(2001)
| Project Number : | 5M01RR000043-410736 |
|---|
| ICD : | NATIONAL CENTER FOR RESEARCH RESOURCES |
|---|
| Project Terms : | cytomegalovirus, dosage, drug screening /evaluation, vaccine development, viral vaccine clinical trial phase I, immunologic memory adult human (19+), clinical research, human subject |
|---|
BONE MARROW DONORS & VOLUNTEERS FOR STUDY OF CMV
(2001)
| Project Number : | 5M01RR000043-410738 |
|---|
| ICD : | NATIONAL CENTER FOR RESEARCH RESOURCES |
|---|
| Project Terms : | bone marrow transplantation, cytomegalovirus, tissue donor, vaccine development, viral vaccine, virus antigen microorganism immunology clinical research, human subject |
|---|
LIPOPEPTIDE VACCINE AGAINST CONSERVED TARGETS OF HIV
(2000)
Abstract :
DESCRIPTION: (Adapted from applicant's abstract): The cellular immune response to HIV has been shown to be robust and continues throughout the natural history of the infection. The capacity for CTL to diminish viral load is substantial, however, sequence variation of HIV limits the effectiveness of the CTL response. Combination pharmacologic therapy against HIV infection known as HAART, has dramatically decreased viral load in many AIDS patients. Despite this control of viral replication, patients who are HAART-responsive for a period of two years or greater, still harbor infectious virus that may, under the appropriate circumstance, become active in replication and cause serious health consequences. The purpose of these studies is to develop immunization strategies in murine models based on immunodominant CTL epitopes from HIV proteins in combination with T-help (HTL) epitopes as part of lipidated peptides. Epitopes will be chosen from sequence-conserved regions of immunogenic HIV proteins. These epitopes will be made into combination vaccines consisting of covalently attached and lipidated HTL+CTL epitopes. The initial approach is to determine the most efficacious means using lipidated vaccines to immunize transgenic mice expressing human HLA Class I and Class II MHC antigens. Mucosal and subcutaneous routes of immunization will be explored. The success of immunization strategies will be evaluated by in vitro methods including the recognition of human antigen presenting cells (APC) which will be infected with different isolates of HIV. The choice of CTL epitope will be made using the criteria that the recognition should be restricted by common HLA Class I alleles. One of the main goals of the proposal and the Innovation Grant Program is to evaluate means to propagate the CTL response using strong T-help (HTL) epitopes. The applicants will evaluate a series of HTL epitopes in combination with selected CTL epitopes using immunization strategies detailed in the proposal. The efficacy of the immune strategies will be examined further using a mouse model of HIV infection referred to as the huPBL-SCID mouse. Immunodeficient mice will be repopulated with human PBL of known HLA type and infected with various isolates of HIV. Vaccine stimulated CTL obtained from transgenic mice will be infused in the previously HIV infected huPBL SCID mice to evaluate their potential for recognition/lysis of infected cells and reduction of viral load. The vaccine strategy that results in the most successful reduction in viral load should be considered in terms of future human immunization clinical protocols for patients on HAART during the time when viral replication is suppressed.
| Project Number : | 1R21AI044313-01 |
|---|
| ICD : | NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES |
|---|
| IRG : | ZAI1 |
|---|
| Project Terms : | AIDS vaccine, HIV infection, cellular immunity, cytotoxic T lymphocyte, nonblood lipoprotein, peptide, vaccine development MHC class I antigen, antigen presenting cell, disease model, drug screening /evaluation, helper T lymphocyte, human immunodeficiency virus, passive immunization, protein sequence, viremia, virus protein, virus replication NOD mouse, SCID mouse, cell line, clinical research, drug administration route, human subject, transgenic animal |
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LIPOPEPTIDE VACCINE W/ ACTIVITY AGAINST HUMAN CYTOMEGALOVIRUS
(2000)
Abstract :
The purpose of this vaccine study is to determine whether a combination vaccine containing a CTl and T-helper epitope, lipidated at the amino terminus will serve to boost the immunity against cytomegalovirus. The protocol will have two phases; the first being amplification of a memory response in seropositive individuals, followed by an immunization of seronegative individuals who have not been productively infected by the virus. To establish safety, a dose escalation study will be done in the group of subjects with prior CMV immunity. The justification for using this group is that they represent a primary target population for an eventual immunization trial in marrow transplant donors. In addition, it is important to establish the safe dosing of this recall immunization, since it is probably more likely to be associated with side effects then the primary immunization. Once a safe dose is established at which an immune recall response occurs, then this dose will be evaluated for safety in primary immunization regimen. Here, CMV nanve subjects will be immunized using a primary injection followed by 2-3 booster injections. Healthy HLA-A*0201-positive male and female volunteers, aged 18-55, who are CMV-seropositive by standard antibody test (see section 4,13,2) will be randomly assigned to one of four dosage groups. Each group will consist of 10 subjects, 4 each will receive an identical dosage of the CMVpp65-A8201 vaccine containing either HTL PADRE peptide (vaccine A) or HTL Tetanus peptide (vaccine B, see Table 2). Within each dose level, 2 subjects will receive placebo, consisting of formulation buffer (i.e., 5-25% DMSO in PBS), and subject assignments will be blinded to the subjects and investigators. Subjects will be stratified by gender for equal distribution into each subgroup. Groups will be immunized sequentially at escalating doses of 50, 250, 1000, 2500 ig/dose. Vaccine A or B or placebo will be administered by injecting 0.5-1.0 ml s.c. into the upper forearm. A booster injection of the same dose and material will be given 28 days after the initial injection. The booster injection will be given in the same anatomical location as the primary inoculation. Subjects will be followed for 12 months for safety observations and immunologic evaluations. In an attempt to study the primary immune response to the CMVpp65-A*0201 vaccine and to establish its safety when used in a multiple booster regimen, HLA A*201/CMV-seronegative volunteers will be immunized using a dose established to be both safe and immunologically efficacious in trial #1. Six subjects will receive a primary immunization followed by a similar booster injection at 28 days and 56 days post-vaccination. There will be no placebo group. If no immunologic response is detected, then no additional booster will be given. However, if a partial or low level response is detected in a subject, then a third booster will be administered between 84-90 days post-primary vaccination. Subjects will be followed for 12 months for safety observations and immunologic evaluations.
| Project Number : | 5M01RR000043-400736 |
|---|
| ICD : | NATIONAL CENTER FOR RESEARCH RESOURCES |
|---|
| Project Terms : | cytomegalovirus, dosage, drug screening /evaluation, vaccine development, viral vaccine clinical trial phase I, immunologic memory adult human (19+), clinical research, human subject |
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BONE MARROW DONORS & VOLUNTEERS FOR STUDY OF CMV
(2000)
Abstract :
Previous results from several laboratories have indicated that the pp65 tegament protein from CMV is the major immunogenic protein recognized by individuals who are seropositive for the virus. CTL clones, which recognize pp65 in the context of ten different HLA Class I alleles, have been identified by our alboratory. The repertoire of epitopes that have been defined for these clones are representative of approximately 95% of the ethnic population of the United sates. Evaluation of whether these CTL epitopes are predominantly used in the general population requires an analysis of the immunogencity of the approach of stimulating peripheral blood lymphocytes form HLA-typed individuals who are seropostive for CMB, in a techique in which the free peptide epitope is added at a high concentration to PBMC in mictowell cultures. We have used this approach with CTL eptiopes specific for pp65 and restricted by HLA A*0201, A*1101,A*2402,A*6901 and B*0702. In each case, we have examined less than five healthy volunteers who are seropositive and hve been shown to respond to the CTL epitopes that are specific for the the HLA allele, which they ecpress. CD8+CTL, which recognize both peptide loaded and CMB infect targets, are amplified 100-foold in a two-week period. The use of the in vitro stimulation procedure allows a sensitive determination of whether an individual is making a CTL response to CMB, and whether pp65 is a component of that response. We wish to demonstrate that a t least five, and ifpossible, ten randomly chosen individualts will respond to a single epitope, suggesting the potential for a universal response to that epitope forma ll individuals who express the same restricting Class I allele. This provides the rationale for an approach to producing vaccine molecules, containing one or more of these epitopes to immunize at risk individuals against CMV infection. The clinical procedures of obtaining both peripheral blood and biopsy are the central part of the lcinical protocol, and could be best carried out under the auspices of the GCRC. We have developed cohorts fo individuals who express the HLA alleles for which we have epitopes form pp65, and more recently, pp150. We wish to carry out these studies, to characterize all of the major eppitopes of CMV pp65 and pp150, which are improtant in the juman immune reaponse to the virus. The prpose for carrying on these studies is to define a peptide-based vaccine that would be applicable to all at-risk individuals. we wil also continue to derive new CMV epitopes, to complete our repertoire of epitopes that would ensure as complete a representation as possible of individuals of different ethnicities that make up the american population. There are several consent forms (A-D) which are active for the protocol.
| Project Number : | 5M01RR000043-400738 |
|---|
| ICD : | NATIONAL CENTER FOR RESEARCH RESOURCES |
|---|
| Project Terms : | bone marrow transplantation, cytomegalovirus, tissue donor, vaccine development, viral vaccine, virus antigen microorganism immunology clinical research, human subject |
|---|
ACQUISITION OF IMMUNITY TO CYTOMEGALOVIRUS
(2000)
Abstract :
CMV infection remains an important problem for transplantation. No successful vaccine has been developed that prevents new infections or even controls existing ones. Pharmacologic treatments have limitations regarding their side effects and the development of resistant strain Asymptomatic CMV-seropositive individuals of survivors of CMV infection after BMT, have an ongoing cytotoxic T lymphocyte (CTL) response to CMV, which can be measured after in vitro stimulation (IVS), either using HLA tetrameter binding or chromium release assay (CRA). The CTL response has multiple targets, however, the response to the tegument protein pp65 predominates in most individuals examined. Although many CTL epitopes have already been mapped from two immunodominant CMV proteins (pp65 and pp150), a significant number have yet been determined, and their elucidation will lead to a broad-based vaccine strategy for all major ethic groups in the United States. Immunodominance of CTL responses to CMV proteins will be determined utilizing IVS strategies together with known CTL epitopes. In contrast, little is known of the predominance of CTL targets from CMV in BMT recipients. The frequency of CMV-specific CTL will be enumerated using flow cytometry with HLA tetramers, and the frequency of those CTL will be compared to viral load measurements using plasma PCR methods. As a means to enhance the immunogenicity of previous defined CTL epitopes, a combinatorial peptide chemistry approach will be used with the known CTL epitopes from pp65 and pp150, which have affinities that are in the micromolar to approach will be used with the known CTL epitopes from pp65 and pp150, which have affinities that are in the micromolar to nanomolar range. Analogue peptides will be defined that will be evaluated initially using HLA A*0201 restricted and pp65- specific T cell clones, to develop a more immunogenic CTL epitope which may lead to a more effective CMV vaccine. The functionality of these analogue peptides will be shown utilizing in vivo immunization of HLA-transgenic mice and IVS studies stimulating a memory response in human PBMC specific for killing of CMV infected fibroblasts in vitro. Two Phase II trials will evaluate modalities including CMV lipopeptide immunization in combination with standard tetanus toxin immunization of BMT recipients. Augmentation of the memory response to tetanus- specific CD4+ T cells will be evaluated by in vitro methods, and the duration of infection-free survival of BMT recipients will be measured using standard clinical parameters. Finally, the utility of targeting a single CMV protein utilizing a CTL epitope vaccine strategy versus targeting two or more CMV proteins will be evaluated using CMV lipopeptide immunization of BMT recipients. Since the endpoint of the trial is a reduction in the incidence of CMV-associated disease, the comparison must be done utilizing a patient population likely to have measurable CMV viral load. Taken together, these strategies of defining CTL epitopes, augmenting their immunogenicity, and defining efficacious ways to delay them to CMT recipients will aid in the development of an effective CMV vaccine both as therapeutic and prophylactic agent.
| Project Number : | 2P01CA030206-190013 |
|---|
| ICD : | NATIONAL CANCER INSTITUTE |
|---|
| Project Terms : | cytomegalovirus, human therapy evaluation, immunity, vaccine development, viral vaccine Herpesviridae disease, bone marrow transplantation, clinical trial phase II /III /IV, cytotoxic T lymphocyte, homologous transplantation, immunologic memory, microorganism immunology, nonhuman therapy evaluation, virus protein clinical research, epitope mapping, flow cytometry, human subject, laboratory mouse, peptide library, transgenic animal |
|---|
LIPOPEPTIDE VACCINE W/ ACTIVITY AGAINST HUMAN CYTOMEGALOVIRUS
(2000)
| Project Number : | 3M01RR000043-40S20736 |
|---|
| ICD : | NATIONAL CENTER FOR RESEARCH RESOURCES |
|---|
| Project Terms : | cytomegalovirus, dosage, drug screening /evaluation, vaccine development, viral vaccine clinical trial phase I, immunologic memory adult human (19+), clinical research, human subject |
|---|
BONE MARROW DONORS & VOLUNTEERS FOR STUDY OF CMV
(2000)
| Project Number : | 3M01RR000043-40S20738 |
|---|
| ICD : | NATIONAL CENTER FOR RESEARCH RESOURCES |
|---|
| Project Terms : | bone marrow transplantation, cytomegalovirus, tissue donor, vaccine development, viral vaccine, virus antigen microorganism immunology clinical research, human subject |
|---|
ACQUISITION OF IMMUNITY TO CYTOMEGALOVIRUS
(1999)
Abstract :
The focus of this proposal is to develop strategies that permit the rapid and efficient transfer of HCMV specific immunity from a bone marrow to an immunosuppressed recipient resulting in protective immune responses that limit or prevent disease. This approach is based on the development of subunit vaccines containing immunogenic viral proteins or peptides. The clear motivation for developing a simple and effective subunit viral protein vaccine is to provide an acceptable alternative to traditional pharmacologic based anti-viral therapy. Our approach will also provide a potentially simpler and cheaper alternative to the approach of adoptive transfer of ex vivo expanded CMV-specific CD8+ CTL from the bone marrow donor into the recipient. Since the HCMV matrix protein pp65 has been previously shown to be the target of a CD8+ CTL response in a large percentage of seropositive individuals, it will be the focus of strategies to define conditions of immunization which result in a widely applicable vaccine preparation. We will utilize the recently developed strategy of defining a minimal cytotoxic epitope (MCE) of a protein, as a means of priming a memory CTL response to the virus that encodes the protein in vitro using peripheral blood of CMV seropositive individuals. Narrowing our search of cytotoxic epitopes specific to several predominant HLAA alleles form HCMV pp65 will permit an early test of the effectiveness of the peptide vaccine strategy. We will determine the ability of an MCE from pp65 to activate memory CTL in a murine model of HCMV recognition. Alternatively, we will also explore whether previously characterized immunologic adjuvants are capable of introducing pp65 full length protein into the Class I protein degradation pathway, thereby causing the development of a primary CD8+ CTL response to the protein in mice. The strength and duration of the CTL response to HCMV pp65 in adjuvant+protein immunized mice will be measured after multiple inoculations of the immunostimulating complex. An adoptive transfer model of CTL immunity to HCMV pp65 will be developed that will combine our two proposed strategies of immunization; adjuvant+protein in the donor, and HLA restricted MCE in the immunosuppressed recipient. HLA-A2.1/Kb transgenic mice will be multiply immunized with adjuvant+protein(pp65), and bone marrow from them will be adoptively transferred into irradiated syngeneic recipients. The recipient's immune response to pp65 will be measured with and without a follow-up peptide immunization. This protocol will closely resemble actual conditions after BMT, and provide a realistic model of the ability of the immunization strategies posed here to provide durable immunity to HCMV in the critical time period after transplant for the immunosuppressed patient. These studies have broad implications for modulation of BMT recipient immunity by manipulation of the donor prior to transplantation, as well as for the general utility of protein vaccines in other groups of patients susceptible to severe HCMV-related disease.
| Project Number : | 5P01CA030206-180013 |
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| ICD : | NATIONAL CANCER INSTITUTE |
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| Project Terms : | cytomegalovirus, drug design /synthesis /production, immunity, microorganism immunology, nonhuman therapy evaluation, viral vaccine cytotoxic T lymphocyte, cytotoxicity, disease model, epitope mapping, gene expression, immunologic memory, leukocyte count, virus protein genetic mapping, laboratory mouse, transgenic animal, western blotting |
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LIPOPEPTIDE VACCINE W/ ACTIVITY AGAINST HUMAN CYTOMEGALOVIRUS
(1999)
Abstract :
The purpose of this vaccine study is to determine whether a combination vaccine containing a CTl and T-helper epitope, lipidated at the amino terminus will serve to boost the immunity against cytomegalovirus. The protocol will have two phases; the first being amplification of a memory response in seropositive individuals, followed by an immunization of seronegative individuals who have not been productively infected by the virus. To establish safety, a dose escalation study will be done in the group of subjects with prior CMV immunity. The justification for using this group is that they represent a primary target population for an eventual immunization trial in marrow transplant donors. In addition, it is important to establish the safe dosing of this recall immunization, since it is probably more likely to be associated with side effects then the primary immunization. Once a safe dose is established at which an immune recall response occurs, then this dose will be evaluated for safety in primary immunization regimen. Here, CMV naove subjects will be immunized using a primary injection followed by 2-3 booster injections. Healthy HLA-A*0201-positive male and female volunteers, aged 18-55, who are CMV-seropositive by standard antibody test (see section 4,13,2) will be randomly assigned to one of four dosage groups. Each group will consist of 10 subjects, 4 each will receive an identical dosage of the CMVpp65-A8201 vaccine containing either HTL PADRE peptide (vaccine A) or HTL Tetanus peptide (vaccine B, see Table 2). Within each dose level, 2 subjects will receive placebo, consisting of formulation buffer (i.e., 5-25% DMSO in PBS), and subject assignments will be blinded to the subjects and investigators. Subjects will be stratified by gender for equal distribution into each subgroup. Groups will be immunized sequentially at escalating doses of 50, 250, 1000, 2500 ig/dose. Vaccine A or B or placebo will be administered by injecting 0.5-1.0 ml s.c. into the upper forearm. A booster injection of the same dose and material will be given 28 days after the initial injection. The booster injection will be given in the same anatomical location as the primary inoculation. Subjects will be followed for 12 months for safety observations and immunologic evaluations. In an attempt to study the primary immune response to the CMVpp65-A*0201 vaccine and to establish its safety when used in a multiple booster regimen, HLA A*201/CMV-seronegative volunteers will be immunized using a dose established to be both safe and immunologically efficacious in trial #1. Six subjects will receive a primary immunization followed by a similar booster injection at 28 days and 56 days post-vaccination. There will be no placebo group. If no immunologic response is detected, then no additional booster will be given. However, if a partial or low level response is detected in a subject, then a third booster will be administered between 84-90 days post-primary vaccination. Subjects will be followed for 12 months for safety observations and immunologic evaluations.
| Project Number : | 5M01RR000043-390736 |
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| ICD : | NATIONAL CENTER FOR RESEARCH RESOURCES |
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| Project Terms : | cytomegalovirus, dosage, drug screening /evaluation, vaccine development, viral vaccine clinical trial phase I, immunologic memory adult human (19+), clinical research, human subject |
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BONE MARROW DONORS & VOLUNTEERS FOR STUDY OF CMV
(1999)
Abstract :
Previous results from several laboratories have indicated that the pp65 tegament protein from CMV is the major immunogenic protein recognized by individuals who are seropositive for the virus. CTL clones, which recognize pp65 in the context of ten different HLA Class I alleles, have been identified by our alboratory. The repertoire of epitopes that have been defined for these clones are representative of approximately 95% of the ethnic population of the United sates. Evaluation of whether these CTL epitopes are predominantly used in the general population requires an analysis of the immunogencity of the approach of stimulating peripheral blood lymphocytes form HLA-typed individuals who are seropostive for CMB, in a techique in which the free peptide epitope is added at a high concentration to PBMC in mictowell cultures. We have used this approach with CTL eptiopes specific for pp65 and restricted by HLA A*0201, A*1101,A*2402,A*6901 and B*0702. In each case, we have examined less than five healthy volunteers who are seropositive and hve been shown to respond to the CTL epitopes that are specific for the the HLA allele, which they ecpress. CD8+CTL, which recognize both peptide loaded and CMB infect targets, are amplified 100-foold in a two-week period. The use of the in vitro stimulation procedure allows a sensitive determination of whether an individual is making a CTL response to CMB, and whether pp65 is a component of that response. We wish to demonstrate that a t least five, and ifpossible, ten randomly chosen individualts will respond to a single epitope, suggesting the potential for a universal response to that epitope forma ll individuals who express the same restricting Class I allele. This provides the rationale for an approach to producing vaccine molecules, containing one or more of these epitopes to immunize at risk individuals against CMV infection. The clinical procedures of obtaining both peripheral blood and biopsy are the central part of the lcinical protocol, and could be best carried out under the auspices of the GCRC. We have developed cohorts fo individuals who express the HLA alleles for which we have epitopes form pp65, and more recently, pp150. We wish to carry out these studies, to characterize all of the major eppitopes of CMV pp65 and pp150, which are important in the human immune reaponse to the virus. The purpose for carrying on these studies is to define a peptide-based vaccine that would be applicable to all at-risk individuals. we wil also continue to derive new CMV epitopes, to complete our repertoire of epitopes that would ensure as complete a representation as possible of individuals of different ethnicities that make up the american population. There are several consent forms (A-D) which are active for the protocol.
| Project Number : | 5M01RR000043-390738 |
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| ICD : | NATIONAL CENTER FOR RESEARCH RESOURCES |
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| Project Terms : | bone marrow transplantation, cytomegalovirus, tissue donor, vaccine development, viral vaccine, virus antigen microorganism immunology clinical research, human subject |
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ACQUISITION OF IMMUNITY TO CYTOMEGALOVIRUS
(1998)
Abstract :
The focus of this proposal is to develop strategies that permit the rapid and efficient transfer of HCMV specific immunity from a bone marrow to an immunosuppressed recipient resulting in protective immune responses that limit or prevent disease. This approach is based on the development of subunit vaccines containing immunogenic viral proteins or peptides. The clear motivation for developing a simple and effective subunit viral protein vaccine is to provide an acceptable alternative to traditional pharmacologic based anti-viral therapy. Our approach will also provide a potentially simpler and cheaper alternative to the approach of adoptive transfer of ex vivo expanded CMV-specific CD8+ CTL from the bone marrow donor into the recipient. Since the HCMV matrix protein pp65 has been previously shown to be the target of a CD8+ CTL response in a large percentage of seropositive individuals, it will be the focus of strategies to define conditions of immunization which result in a widely applicable vaccine preparation. We will utilize the recently developed strategy of defining a minimal cytotoxic epitope (MCE) of a protein, as a means of priming a memory CTL response to the virus that encodes the protein in vitro using peripheral blood of CMV seropositive individuals. Narrowing our search of cytotoxic epitopes specific to several predominant HLAA alleles form HCMV pp65 will permit an early test of the effectiveness of the peptide vaccine strategy. We will determine the ability of an MCE from pp65 to activate memory CTL in a murine model of HCMV recognition. Alternatively, we will also explore whether previously characterized immunologic adjuvants are capable of introducing pp65 full length protein into the Class I protein degradation pathway, thereby causing the development of a primary CD8+ CTL response to the protein in mice. The strength and duration of the CTL response to HCMV pp65 in adjuvant+protein immunized mice will be measured after multiple inoculations of the immunostimulating complex. An adoptive transfer model of CTL immunity to HCMV pp65 will be developed that will combine our two proposed strategies of immunization; adjuvant+protein in the donor, and HLA restricted MCE in the immunosuppressed recipient. HLA-A2.1/Kb transgenic mice will be multiply immunized with adjuvant+protein(pp65), and bone marrow from them will be adoptively transferred into irradiated syngeneic recipients. The recipient's immune response to pp65 will be measured with and without a follow-up peptide immunization. This protocol will closely resemble actual conditions after BMT, and provide a realistic model of the ability of the immunization strategies posed here to provide durable immunity to HCMV in the critical time period after transplant for the immunosuppressed patient. These studies have broad implications for modulation of BMT recipient immunity by manipulation of the donor prior to transplantation, as well as for the general utility of protein vaccines in other groups of patients susceptible to severe HCMV-related disease.
| Project Number : | 5P01CA030206-170013 |
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| ICD : | NATIONAL CANCER INSTITUTE |
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| Project Terms : | cytomegalovirus, drug design /synthesis /production, immunity, microorganism immunology, nonhuman therapy evaluation, viral vaccine cytotoxic T lymphocyte, cytotoxicity, disease model, epitope mapping, gene expression, immunologic memory, leukocyte count, virus protein genetic mapping, laboratory mouse, transgenic animal, western blotting |
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ACQUISITION OF IMMUNITY TO CYTOMEGALOVIRUS
(1996)
Abstract :
The focus of this proposal is to develop strategies that permit the rapid and efficient transfer of HCMV specific immunity from a bone marrow to an immunosuppressed recipient resulting in protective immune responses that limit or prevent disease. This approach is based on the development of subunit vaccines containing immunogenic viral proteins or peptides. The clear motivation for developing a simple and effective subunit viral protein vaccine is to provide an acceptable alternative to traditional pharmacologic based anti-viral therapy. Our approach will also provide a potentially simpler and cheaper alternative to the approach of adoptive transfer of ex vivo expanded CMV-specific CD8+ CTL from the bone marrow donor into the recipient. Since the HCMV matrix protein pp65 has been previously shown to be the target of a CD8+ CTL response in a large percentage of seropositive individuals, it will be the focus of strategies to define conditions of immunization which result in a widely applicable vaccine preparation. We will utilize the recently developed strategy of defining a minimal cytotoxic epitope (MCE) of a protein, as a means of priming a memory CTL response to the virus that encodes the protein in vitro using peripheral blood of CMV seropositive individuals. Narrowing our search of cytotoxic epitopes specific to several predominant HLAA alleles form HCMV pp65 will permit an early test of the effectiveness of the peptide vaccine strategy. We will determine the ability of an MCE from pp65 to activate memory CTL in a murine model of HCMV recognition. Alternatively, we will also explore whether previously characterized immunologic adjuvants are capable of introducing pp65 full length protein into the Class I protein degradation pathway, thereby causing the development of a primary CD8+ CTL response to the protein in mice. The strength and duration of the CTL response to HCMV pp65 in adjuvant+protein immunized mice will be measured after multiple inoculations of the immunostimulating complex. An adoptive transfer model of CTL immunity to HCMV pp65 will be developed that will combine our two proposed strategies of immunization; adjuvant+protein in the donor, and HLA restricted MCE in the immunosuppressed recipient. HLA-A2.1/Kb transgenic mice will be multiply immunized with adjuvant+protein(pp65), and bone marrow from them will be adoptively transferred into irradiated syngeneic recipients. The recipient's immune response to pp65 will be measured with and without a follow-up peptide immunization. This protocol will closely resemble actual conditions after BMT, and provide a realistic model of the ability of the immunization strategies posed here to provide durable immunity to HCMV in the critical time period after transplant for the immunosuppressed patient. These studies have broad implications for modulation of BMT recipient immunity by manipulation of the donor prior to transplantation, as well as for the general utility of protein vaccines in other groups of patients susceptible to severe HCMV-related disease.
| Project Number : | 5P01CA030206-150013 |
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| ICD : | NATIONAL CANCER INSTITUTE |
|---|
| Project Terms : | cytomegalovirus, drug design /synthesis /production, immunity, microorganism immunology, nonhuman therapy evaluation, viral vaccine cytotoxic T lymphocyte, cytotoxicity, disease model, epitope mapping, gene expression, immunologic memory, leukocyte count, virus protein genetic mapping, laboratory mouse, transgenic animal, western blotting |
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ACQUISITION OF IMMUNITY TO CYTOMEGALOVIRUS
(1995)
Abstract :
The focus of this proposal is to develop strategies that permit the rapid and efficient transfer of HCMV specific immunity from a bone marrow to an immunosuppressed recipient resulting in protective immune responses that limit or prevent disease. This approach is based on the development of subunit vaccines containing immunogenic viral proteins or peptides. The clear motivation for developing a simple and effective subunit viral protein vaccine is to provide an acceptable alternative to traditional pharmacologic based anti-viral therapy. Our approach will also provide a potentially simpler and cheaper alternative to the approach of adoptive transfer of ex vivo expanded CMV-specific CD8+ CTL from the bone marrow donor into the recipient. Since the HCMV matrix protein pp65 has been previously shown to be the target of a CD8+ CTL response in a large percentage of seropositive individuals, it will be the focus of strategies to define conditions of immunization which result in a widely applicable vaccine preparation. We will utilize the recently developed strategy of defining a minimal cytotoxic epitope (MCE) of a protein, as a means of priming a memory CTL response to the virus that encodes the protein in vitro using peripheral blood of CMV seropositive individuals. Narrowing our search of cytotoxic epitopes specific to several predominant HLAA alleles form HCMV pp65 will permit an early test of the effectiveness of the peptide vaccine strategy. We will determine the ability of an MCE from pp65 to activate memory CTL in a murine model of HCMV recognition. Alternatively, we will also explore whether previously characterized immunologic adjuvants are capable of introducing pp65 full length protein into the Class I protein degradation pathway, thereby causing the development of a primary CD8+ CTL response to the protein in mice. The strength and duration of the CTL response to HCMV pp65 in adjuvant+protein immunized mice will be measured after multiple inoculations of the immunostimulating complex. An adoptive transfer model of CTL immunity to HCMV pp65 will be developed that will combine our two proposed strategies of immunization; adjuvant+protein in the donor, and HLA restricted MCE in the immunosuppressed recipient. HLA-A2.1/Kb transgenic mice will be multiply immunized with adjuvant+protein(pp65), and bone marrow from them will be adoptively transferred into irradiated syngeneic recipients. The recipient's immune response to pp65 will be measured with and without a follow-up peptide immunization. This protocol will closely resemble actual conditions after BMT, and provide a realistic model of the ability of the immunization strategies posed here to provide durable immunity to HCMV in the critical time period after transplant for the immunosuppressed patient. These studies have broad implications for modulation of BMT recipient immunity by manipulation of the donor prior to transplantation, as well as for the general utility of protein vaccines in other groups of patients susceptible to severe HCMV-related disease.
| Project Number : | 2P01CA030206-140013 |
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| ICD : | NATIONAL CANCER INSTITUTE |
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| Project Terms : | cytomegalovirus, drug design /synthesis /production, immunity, microorganism immunology, nonhuman therapy evaluation, viral vaccine cytotoxic T lymphocyte, cytotoxicity, disease model, epitope mapping, gene expression, immunologic memory, leukocyte count, virus protein genetic mapping, laboratory mouse, transgenic animal, western blotting |
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PROTEINS BINDING TO THE HUMAN GAMMA INTERFERON PROMOTER
(1994)
Abstract :
The product of the Interferon gamma (IFN-gamma) gene is a potent modulator of the immune response. Induction of both IFN-gamma and IL2 mRNA transcription is concurrently stimulated by T-cell activation agents. Molecular analysis of the human IFN-gamma promoter has revealed separate sequence elements which either respond to T-cell activation signals, or to an alternate activator encoded by the human retroviruses, HTLV-I and II. Using a combination of gel retardation analysis, methylation protection, and UV cross-linking we will identify the molecular components in the nucleus which bind to the previously identified functionally important sequences within the IFN-gamma promoter. Since human retroviruses are capable of being transcriptionally activated by T-cell inducing agents, it is likely that the nuclear products identified by the above techniques which bind to sequences in the IFN-gamma promoter would also interact with the products of the viral transactivator genes (tax I or tax II) similar to the protein:promoter binding of the AP-1 and Fos gene products. Some of these proteins (such as Nuclear Factor-kB) may also be utilized by the HIV promoter within the long terminal repeat, as a T-cell activation-specific transcription factor. We will isolate the genes which encode the nuclear factors important in IFN-gamma regulation, by screening an expression library with either an oligonucleotide specific to a nuclear factor binding site, or an oligonucleotide derived from the isolated and sequenced nuclear factors. These studies are aimed at an understanding of how the viral activation of the IFN-gamma gene is related to the concurrent oncogenic transformation of cells harboring the HTLV type of retrovirus. Use of Jurkat cells as a model system is one approach through the introduction of vectors expressing tax and studying their effects on endogenous genes such as IFN-gamma. Introduction of tax expression vectors and/or IFN-gamma promoter deletion plasmids into peripheral blood lymphocytes by electroporation would be an additional physiological approach to characterize sequences important in promoter activation, as compared to the Jurkat T-cell tumor model. Additional studies have revealed that the immunosuppressive agent cyclosporine A (Cs A) can overcome the activation of the IFN-gamma promoter by T-cell activating agents. It is an important goal to determine at which level of regulation does Cs A inhibit IFN-gamma transcription. Since tax gene products are able to relieve the inhibitory effects of Cs A on IFN-gamma induction, it will be equally important to discover how those two pathways interact to modify IFN-gamma gene regulation. The discovery that HTLV gene products modify the action of Cs A on the immune system has potential implication for patients in need of immunosuppressive therapy (transplantation) who have attendant retroviral- associated (HTLV) disease.
| Project Number : | 5R01CA052177-03 |
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| ICD : | NATIONAL CANCER INSTITUTE |
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| IRG : | EI |
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| Project Terms : | DNA binding protein, genetic promoter element, genetic transcription, human T lymphotropic virus 1, human T lymphotropic virus 2, interferon, leukocyte activation /transformation T lymphocyte, crosslink, interferon inducer, methylation, virus host interaction cyclosporine, gel mobility shift assay, genetic library, genetic manipulation, laboratory mouse, laboratory rabbit, molecular cloning, mutation, transfection, ultraviolet radiation |
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ANTIGEN BINDING SITES OF HUMAN T CELL RECEPTORS
(1993)
Abstract :
The product of the Interferon gamma (IFN-gamma) gene is a potent modulator of the immune response. Induction of both IFN-gamma and IL2 mRNA transcription is concurrently stimulated by T-cell activation agents. Molecular analysis of the human IFN-gamma promoter has revealed separate sequence elements which either respond to T-cell activation signals, or to an alternate activator encoded by the human retroviruses, HTLV-I and II. Using a combination of gel retardation analysis, methylation protection, and UV cross-linking we will identify the molecular components in the nucleus which bind to the previously identified functionally important sequences within the IFN-gamma promoter. Since human retroviruses are capable of being transcriptionally activated by T-cell inducing agents, it is likely that the nuclear products identified by the above techniques which bind to sequences in the IFN-gamma promoter would also interact with the products of the viral transactivator genes (tax I or tax II) similar to the protein:promoter binding of the AP-1 and Fos gene products. Some of these proteins (such as Nuclear Factor-kB) may also be utilized by the HIV promoter within the long terminal repeat, as a T-cell activation-specific transcription factor. We will isolate the genes which encode the nuclear factors important in IFN-gamma regulation, by screening an expression library with either an oligonucleotide specific to a nuclear factor binding site, or an oligonucleotide derived from the isolated and sequenced nuclear factors. These studies are aimed at an understanding of how the viral activation of the IFN-gamma gene is related to the concurrent oncogenic transformation of cells harboring the HTLV type of retrovirus. Use of Jurkat cells as a model system is one approach through the introduction of vectors expressing tax and studying their effects on endogenous genes such as IFN-gamma. Introduction of tax expression vectors and/or IFN-gamma promoter deletion plasmids into peripheral blood lymphocytes by electroporation would be an additional physiological approach to characterize sequences important in promoter activation, as compared to the Jurkat T-cell tumor model. Additional studies have revealed that the immunosuppressive agent cyclosporine A (Cs A) can overcome the activation of the IFN-gamma promoter by T-cell activating agents. It is an important goal to determine at which level of regulation does Cs A inhibit IFN-gamma transcription. Since tax gene products are able to relieve the inhibitory effects of Cs A on IFN-gamma induction, it will be equally important to discover how those two pathways interact to modify IFN-gamma gene regulation. The discovery that HTLV gene products modify the action of Cs A on the immune system has potential implication for patients in need of immunosuppressive therapy (transplantation) who have attendant retroviral- associated (HTLV) disease.
| Project Number : | 1R01CA052177-01 |
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| ICD : | NATIONAL CANCER INSTITUTE |
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| IRG : | EI |
|---|
| Project Terms : | GENETICS, GENES, REGULATORY SEQUENCES, PROMOTERS, GENETICS, GENETIC REGULATION, TRANSCRIPTION, IMMUNITY, CELLULAR, LEUKOCYTE ACTIVATION, TRANSFORMATION AND PROLIFERATION, IMMUNITY, CYTOKINES, LYMPHOKINES, INTERFERONS, PROTEINS, BINDING PROTEINS, DNA-BINDING PROTEINS, VIRUSES, RETROVIRIDAE, HUMAN T-CELL LEUKEMIA-LYMPHOMA VIRUS1, VIRUSES, RETROVIRIDAE, HUMAN T-CELL LEUKEMIA-LYMPHOMA VIRUS2 ALKYL (GROUPS), METHYLATION-DEMETHYLATION, BLOOD CELLS, T LYMPHOCYTES, CHEMICAL BONDS, CROSSLINKS, IMMUNOLOGY, INTERFERON INDUCERS, VIRUS DISEASE CHARACTERISTICS, HOST-VIRUS ANIMALS, CHORDATES, MAMMALS, LAGOMORPHS, RABBITS (LABORATORY), ANIMALS, CHORDATES, MAMMALS, RODENTS, MYOMORPHA, MICE (LABORATORY), ANTIBIOTICS, CYCLOSPORIN A, GENETIC MANIPULATION, TRANSFECTION, GENETICS, BIOCHEMICAL GENETICS, MOLECULAR CLONING, GENETICS, GENETIC LIBRARIES, GENETICS, MUTATION, PHYSICAL SEPARATION, ELECTROPHORESIS, GEL, RADIATION, ELECTROMAGNETIC WAVES, ULTRAVIOLET RAYS (290NM TO 380NM), genetic manipulation |
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PROTEINS BINDING TO THE HUMAN GAMMA INTERFERON PROMOTER
(1993)
Abstract :
The product of the Interferon gamma (IFN-gamma) gene is a potent modulator of the immune response. Induction of both IFN-gamma and IL2 mRNA transcription is concurrently stimulated by T-cell activation agents. Molecular analysis of the human IFN-gamma promoter has revealed separate sequence elements which either respond to T-cell activation signals, or to an alternate activator encoded by the human retroviruses, HTLV-I and II. Using a combination of gel retardation analysis, methylation protection, and UV cross-linking we will identify the molecular components in the nucleus which bind to the previously identified functionally important sequences within the IFN-gamma promoter. Since human retroviruses are capable of being transcriptionally activated by T-cell inducing agents, it is likely that the nuclear products identified by the above techniques which bind to sequences in the IFN-gamma promoter would also interact with the products of the viral transactivator genes (tax I or tax II) similar to the protein:promoter binding of the AP-1 and Fos gene products. Some of these proteins (such as Nuclear Factor-kB) may also be utilized by the HIV promoter within the long terminal repeat, as a T-cell activation-specific transcription factor. We will isolate the genes which encode the nuclear factors important in IFN-gamma regulation, by screening an expression library with either an oligonucleotide specific to a nuclear factor binding site, or an oligonucleotide derived from the isolated and sequenced nuclear factors. These studies are aimed at an understanding of how the viral activation of the IFN-gamma gene is related to the concurrent oncogenic transformation of cells harboring the HTLV type of retrovirus. Use of Jurkat cells as a model system is one approach through the introduction of vectors expressing tax and studying their effects on endogenous genes such as IFN-gamma. Introduction of tax expression vectors and/or IFN-gamma promoter deletion plasmids into peripheral blood lymphocytes by electroporation would be an additional physiological approach to characterize sequences important in promoter activation, as compared to the Jurkat T-cell tumor model. Additional studies have revealed that the immunosuppressive agent cyclosporine A (Cs A) can overcome the activation of the IFN-gamma promoter by T-cell activating agents. It is an important goal to determine at which level of regulation does Cs A inhibit IFN-gamma transcription. Since tax gene products are able to relieve the inhibitory effects of Cs A on IFN-gamma induction, it will be equally important to discover how those two pathways interact to modify IFN-gamma gene regulation. The discovery that HTLV gene products modify the action of Cs A on the immune system has potential implication for patients in need of immunosuppressive therapy (transplantation) who have attendant retroviral- associated (HTLV) disease.
| Project Number : | 5R01CA052177-02 |
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| ICD : | NATIONAL CANCER INSTITUTE |
|---|
| IRG : | EI |
|---|
| Project Terms : | DNA binding protein, genetic promoter element, genetic transcription, human T lymphotropic virus 1, human T lymphotropic virus 2, interferon, leukocyte activation /transformation T lymphocyte, crosslink, interferon inducer, methylation, virus host interaction cyclosporine, genetic manipulation, genomic library, laboratory mouse, laboratory rabbit, molecular cloning, mutation, transfection, ultraviolet radiation |
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