Hatch W C, Tanaka K E, Calvelli T, Rashbaum W K, Kress Y, Lyman W D
Department of Pathology, Pediatrics, and Obstetrics & Gynecology, Albert Einstein College of Medicine, Bronx, NY 10461.
J Immunol. 1992 May 15;148(10):3055-61.
Human fetal thymuses were obtained from abortuses of HIV-1 seronegative females. Thymocytes were isolated and cultured for 2 days with PHA. Thereafter, the culture was divided and half of the cells were exposed to the HIV-1 RF isolate for 4 h. After this incubation period, the HIV-1 exposed and nonexposed control cells were cultured in RPMI 1640 supplemented with IL-2 for 30 days and subsequently maintained in RPMI without the addition of growth factors. Long term culture of both HIV-1 exposed and control thymocytes has yielded two cell lines that have been maintained for more than 3 yr without the addition of growth factors. Flow cytometry using mAb that recognize T cell differentiation markers was used to analyze cell phenotypes. The HIV-1 exposed thymocyte cell line (E88/RF) was shown to be HIV-1 infected by p24 ELISA, reverse transcriptase activity, immunocytochemistry, in situ hybridization, polymerase chain reaction, electron microscopy, and to produce infectious particles by a syncytial forming assay. The non-HIV-1-exposed thymocyte cell line (T412) has remained negative by all criteria for HIV-1 infection. Flow cytometry showed the T412 cells to be positive for the T cell markers CD45, CD38, and CD4 but negative for all other markers tested. The E88/RF cells are positive for CD45 and CD38 but negative for CD4 and other markers. These data report the isolation of two human fetal thymocyte cell lines; one uninfected and susceptible to HIV-1 infection, and the other persistently and productively infected with HIV-1 with little cytopathology. These findings suggest that HIV-1 can persistently infect early T cells and may alter T cell differentiation.
人类胎儿胸腺取自HIV-1血清阴性女性的流产胎儿。分离胸腺细胞并用PHA培养2天。此后，将培养物分开，一半细胞暴露于HIV-1 RF毒株4小时。在此孵育期后，将暴露于HIV-1和未暴露的对照细胞在补充有IL-2的RPMI 1640中培养30天，随后在不添加生长因子的RPMI中维持培养。对暴露于HIV-1的胸腺细胞和对照胸腺细胞进行长期培养，得到了两个细胞系，在不添加生长因子的情况下已维持培养超过3年。使用识别T细胞分化标志物的单克隆抗体进行流式细胞术分析细胞表型。通过p24 ELISA、逆转录酶活性、免疫细胞化学、原位杂交、聚合酶链反应、电子显微镜检测，显示暴露于HIV-1的胸腺细胞系（E88/RF）被HIV-1感染，并通过合胞体形成试验产生感染性颗粒。未暴露于HIV-1的胸腺细胞系（T412）在所有HIV-1感染标准下均为阴性。流式细胞术显示T412细胞的T细胞标志物CD45、CD38和CD4呈阳性，但对所有其他检测标志物呈阴性。E88/RF细胞的CD45和CD38呈阳性，但CD4和其他标志物呈阴性。这些数据报告了两个人类胎儿胸腺细胞系的分离；一个未感染且易受HIV-1感染，另一个持续且高效地感染HIV-1，几乎没有细胞病理学变化。这些发现表明HIV-1可持续感染早期T细胞，并可能改变T细胞分化。
