权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Epidemiology of antibody against reverse transcriptase of AIDS virus

艾滋病病毒逆转录酶抗体的流行病学

基本信息

批准号：
01480196
负责人：
HOSHINO Hiroo
金额：
$ 3.39万
依托单位：
Gunma University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (B)
财政年份：
1989
资助国家：
日本
起止时间：
1989 至 1990
项目状态：
已结题

项目摘要

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.

为了分离产生大量HIV-1的细胞系，用于逆转录酶（RT）的纯化，我们分离了11株主要来自日本血友病患者的HIV-1株。这些HIV-1被转移到人类T细胞系或单核细胞系。MOLT4/HIV- 1[GUN-4]细胞大量产生HIV-1。我们进一步从它们身上分离出细胞克隆，它们产生了大量的HIV-1。利用lambda噬菌体分离HIV-1[GUN-1]的分子克隆，通过基因工程产生HIV-1 RT。通过将这些噬菌体克隆转染人T细胞，证实了HIV-1分子克隆的感染性。这些无性系的Pol基因可用于生产。我们建立了一种新的检测HIV-1抗RT抗体的检测系统：抗体抗HIV-1 RT在56℃热失活后稳定酶。HIV-1阳性受试者的血清特异性稳定HIV-1 RT，但不稳定HIV-2、小鼠白血病病毒、人t细胞白血病病毒1型或牛白血病病毒的RT。使用日本、英国或加拿大hiv -1阳性受试者的血清研究稳定抗体存在的临床意义。稳定法比常规中和法更频繁地检测到抗HIV-1 RT抗体。血清稳定活性与外周血cd4阳性细胞数量呈显著负相关。HIV-1 RT的稳定试验可以用低至10mu1的血清来完成。该试验与中和试验一起被证明对估计HIV-1感染的严重程度和预后具有临床意义。