| Project/Area Number |
01480196
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| Research Category |
Grant-in-Aid for General Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
Hygiene
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| Research Institution | Gunma University |
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Principal Investigator |
HOSHINO Hiroo Department of Hygiene, Gunma University School of Medicine, Professor, 医学部・衛生学教室, 教授 (00107434)
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| Co-Investigator(Kenkyū-buntansha) |
YAMAMOTO Akiko Department of Hygiene, Gunma University School of Medicine, Assistant, 医学部・衛生学教室, 助手 (70092012)
TAKEUCHI Yasuhiro Department of Hygiene, Gunma University School of Medicine, Assistant, 医学部・衛生学教室, 助手 (70188168)
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| Project Period (FY) |
1989 – 1990
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| Project Status |
Completed (Fiscal Year 1991)
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| Budget Amount *help |
¥5,300,000 (Direct Cost: ¥5,300,000)
Fiscal Year 1990: ¥1,800,000 (Direct Cost: ¥1,800,000)
Fiscal Year 1989: ¥3,500,000 (Direct Cost: ¥3,500,000)
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| Keywords | AIDS / HIV-1 / Reverse transcriptase / Antibody / AIDS(Acquired immunedeficieny Syndromel) / HIV(Human immunodeficiency virus) |
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| Research Abstract |
To isolate cell lines producing a large amount of HIV-1, which is to be used for purification of reverse transcriptase (RT), we have isolated 11 HIV-1 strans mainly derived from Japanese hemophiliacs. These HIV-1 were transferred to human T cell lines or monocytic cell lines. MOLT4/HIV- 1[GUN-4]cells produced HIV-1 abundantly. Further we isolated cell clones from them, which were producing a large amount of HIV-1.
To produce HIV-1 RT by genetic engineering, molecular clones of HIV-1[GUN-1]were isolated using lambda phages. Infectivities of molecular clones of HIV-1 was confirmed by transfecting these phage clone into human T cells. Pol gene of these clones can be used for production of. recombinant HIV-1 RT.
We have developed a new assay system for detection of antibody against RT of HIV-1 : Antibody against HIV-l RT stabilized the enzyme upon its heat inactivation at 56ﾟC. Sera from HIV-1positive subjects specifically stabilized HIV-1 RT, but not RTs of HIV-2, murine leukemia virus, human-n T-cell leukemia virus type 1 or bovine leukemia virus.
The clinical significance of the presence of stabilizing antibodies was studied using sera of Japanese, British or Canadian HIV-1-positive subjects. Antibody against HIV-1 RT was more frequently detected by stabilization assay than conventional neutralization assay. There was a significant inverse correlation between stabilizing activities of sera and numbers of CD4-positive cells in peripheral blood. The stabilization assay of HIV-1 RT can be done using as small as 10mu1 of serum. This assay together with neutralization assay was shown to have clinical significance to estimate the severity and prognosis of HIV-1 infection.
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