| Project/Area Number |
14207013
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| Research Category |
Grant-in-Aid for Scientific Research (A)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Virology
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| Research Institution | The University of Tokyo |
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Principal Investigator |
IWAMOTO Aikichi The University of Tokyo, Institute of Medical Science, Professor, 医科学研究所, 教授 (10133076)
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| Co-Investigator(Kenkyū-buntansha) |
ODAWARA Takashi University of Tokyo, Institute of Medical Science, Lecturer, 医科学研究所, 講師 (40204218)
NAKAMURA Tetsuya University of Tokyo, Institute of Medical Science, Associate Professor, 医科学研究所, 助教授 (30189047)
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| Project Period (FY) |
2002 – 2003
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| Project Status |
Completed (Fiscal Year 2003)
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| Budget Amount *help |
¥51,090,000 (Direct Cost: ¥39,300,000、Indirect Cost: ¥11,790,000)
Fiscal Year 2003: ¥14,690,000 (Direct Cost: ¥11,300,000、Indirect Cost: ¥3,390,000)
Fiscal Year 2002: ¥36,400,000 (Direct Cost: ¥28,000,000、Indirect Cost: ¥8,400,000)
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| Keywords | HIV / vaccine / epitope / CTL / tetramer |
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| Research Abstract |
(1)Analysis of CTL epitopes
HIV derived from HLA A24-positive or negative patients was sequenced. We found that one of the HLA A24-restricted CTL epitopes in Nef region (Nef138-10) had a consistent nucleotide mutation in HLA A24-positive patients, but not in HLA A24-negative patients. The observation suggests that immunological pressure caused by CTLs selected the mutation in Nef138-10. The observations that we obtained will be published in Journal of Virology.
(2)Phase I clinical study using HIV vaccine
We started a phase I clinical study to investigate whether HIV-specific immunity augmented by HIV vaccine can control HIV proliferation in HIV-infected patients. For the therapeutic vaccine, we used autologous dendritic cells pulsed with several HIV peptides corresponding to CTL epitopes. Four patients were enrolled in the study, and two finished vaccine administration. Immunological analysis is now under way.
(3)Genetic analysis of CTL using micro array system
Although HIV-specific CTL can be sorted by flowcytometer using tetramer staining, the expected number of CTLs is small. Thus, mRNA must be efficiently amplified before micro array assay. We are now working to develop not only efficient but also reproducible method for amplification using T7 polymerase.
(4)Analysis of CTL epitope using overlapping peptides that span whole HIV sequence.
We made approximately 600 peptides (15 amino-acid-length peptides which overlap neighboring peptides by 10 aa) that span whole HIV sequence. Peripheral blood mononuclear cells derived from HIV-infected patients were cultured with pooled peptides (10〜,50 peptides), and IFN-r-secreting lymphocytes were analyzed using ELISPOT assay. By reducing the number of peptides in each pool, we are now narrowing peptides that serve strong CTL epitope activity.
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