El Messaoudi K, Thiry L, Van Tieghem N, Liesnard C, Englert Y, Moguilevsky N, Bollen A
Department of Applied Genetics, Faculty of Sciences, Erasme Hospital, Université Libre de Bruxelles, Nivelles, Belgium.
AIDS. 1999 Feb 25;13(3):333-9. doi: 10.1097/00002030-199902250-00005.
To investigate HIV-1 infectivity in the natural environment of vaginal secretions.
Vaginal wash samples collected from 14 healthy women were incubated in vitro with various HIV-1 strains for 10 min at 37 degrees C and then assayed for infectivity on primary lymphocyte cultures, or on CEM cells, or on CD4- ME180 cells derived from vaginal epithelium.
HIV-1 infectivity was measured by early virus growth in the various host cells tested using a quantitative p24 assay and by the Karber procedure.
Preincubation of HIV-1(IIIB) with vaginal wash samples or 2 microg/ml cathepsin D increased the ability of the virus to grow in lymphocyte cultures. The vaginal wash effect was abolished by 5 microg/ml pepstatin A, an inhibitor of aspartyl proteases. Presence of precursor and mature forms of cathepsin D in vaginal wash was demonstrated after passage through a pepstatin A-agarose column. Median tissue culture infective doses of HIV-1(IIIB) and HIV-1(JRFL) strains were increased 14.4-fold and 18-fold, respectively, after preincubation in vaginal wash sample, and were increased by pretreatment with 2 microg/ml cathepsin D. When CD4 receptors of CEMss cells were blocked by OKT4a monoclonal antibody, the cells lost susceptibility to HIV-1 (IIIB), but supported the growth of virus pretreated with vaginal wash sample or cathepsin D. These treated viruses were able to initiate infection of CD4-ME180 epithelial cells, which were not receptive to untreated virus. ME180 cells were shown to possess the messenger of CXC-chemokine receptor-4.
Vaginal secretions may help HIV-1 transmission to women by increasing infectivity for CD4+ cells and allowing entrance into some CD4-epithelial cells.
研究HIV-1在阴道分泌物自然环境中的感染性。
从14名健康女性收集的阴道冲洗样本在体外与各种HIV-1毒株于37℃孵育10分钟，然后检测其对原代淋巴细胞培养物、CEM细胞或源自阴道上皮的CD4-ME180细胞的感染性。
通过使用定量p24测定法和Karber程序在各种测试宿主细胞中早期病毒生长来测量HIV-1感染性。
HIV-1（IIIB）与阴道冲洗样本或2μg/ml组织蛋白酶D预孵育增加了病毒在淋巴细胞培养物中的生长能力。5μg/ml胃蛋白酶抑制剂A（一种天冬氨酸蛋白酶抑制剂）消除了阴道冲洗的作用。通过胃蛋白酶抑制剂A-琼脂糖柱后，在阴道冲洗液中证实了组织蛋白酶D的前体和成熟形式的存在。HIV-1（IIIB）和HIV-1（JRFL）毒株的半数组织培养感染剂量在阴道冲洗样本中预孵育后分别增加了14.4倍和18倍，并且通过用2μg/ml组织蛋白酶D预处理而增加。当CEMss细胞的CD4受体被OKT4a单克隆抗体阻断时，细胞对HIV-1（IIIB）失去敏感性，但支持用阴道冲洗样本或组织蛋白酶D预处理的病毒生长。这些经处理的病毒能够引发对未处理病毒不敏感的CD4-ME180上皮细胞的感染。ME180细胞被证明具有CXC趋化因子受体-4的信使。
阴道分泌物可能通过增加对CD4+细胞的感染性并允许进入一些CD4-上皮细胞来帮助HIV-1传播给女性。
