Epidemiology of antibody against reverse transcriptase of AIDS virus

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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.