1991 Fiscal Year Final Research Report Summary
Development of Vaccine for Preventing HTLV-I and HIV Infection and Disease Development Using Vaccinie Virus Vector
| Project/Area Number |
01870022
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| Research Category |
Grant-in-Aid for Developmental Scientific Research
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| Allocation Type | Single-year Grants |
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| Research Field |
Virology
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| Research Institution | Kyoto University |
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Principal Investigator |
SHIDA Hisatoshi Inst, for Virus Res. Kyoto Univ. Associate Professor, ウイルス研究所, 助教授 (00144395)
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| Co-Investigator(Kenkyū-buntansha) |
KANNAGI Mari Medical School Kumamoto Univ. Assistant Professor, 医学部, 助手 (80202034)
TANAKA Yuetsu Hygenic Sci. Kitasato Univ. Assistant Professor, 衛生学部, 助手 (30163588)
INOKO Hidetoshi Medical School Tokai Univ. Associate Professor, 医学部, 助教授 (10101932)
SHIKU Hiroshi Medical School Nagasaki Univ. Professor, 医学部, 教授 (80154194)
HAYAMI Masanori Inst. for Virus Res. Kyoto Univ. Professor, ウイルス研究所, 教授 (40072946)
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| Project Period (FY) |
1989 – 1991
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| Keywords | HTLV-I / Vaccine Vector / Vaccine / HAM / CTL / SIV |
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| Research Abstract |
(1) We examined whether there is deference between CTL activities of healthy carriers and that of ATL or HAM patients. It has been found that the HAM patients had CTL against Tax protein whereas the healthy carriers did not. Epitope mapping using synthesized peptides revealed the antigenicity of amino terminal of Tax. These CTLs were restricted by HLAA2. Measurement of. priming of T cells revealed that among healthy carriers some had primed T cel-is to pX proteins (Tax, Rex, and p2l^X) and the other did not. These high responses correlated to the HLA types characteristic to HAM patients. The primed T cells had CD8 antigen. These results suggest the relationship between anti Tax CTL and HAM.
(2) To identify the neutralization epitope of HTLV-I, anti env MAbs which had the neutralizing activity were produced. Epitope mapping using synthetic peptides revealed the gp46 amino acid sequence 191-196 as the binding site of the MAb. Immunization of the rabbits by this peptide elicited neutralizing antibody and prevented HTLV-I infection.
(3) To construct more efficient RW vectors, we introduced mutations into the region of p7.5 promoter and then placed them in tandemly repeated position. Furthermore we combined them with ATI promoter to produce stronger promoter. When we measure its efficacy by expression of CAT gene as a reporter, CAT protein was synthesized whose amount was approximately 5% of that of total cell proteins.
(4) We constructed the RW which harbors the SIV env gene under the control of ATI hybrid promoter. After cynomologous monkeys were immunized with this RVV, they were challenged by SIV to examine its capability of preventing infection. All three monkeys challenged were infected by SIV. In the second experiment, partially-purified env protein was injected as booster after the immunization by the RW, and then the monkeys were challenged by SIV. One of three monkeys immunized was protected.
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