Eylar E H, Lefranc C, Báez I, Colón-Martinez S L, Yamamura Y, Rodriguez N, Yano N, Breithaupt T B
Ponce School of Medicine, Department of Biochemistry, Puerto Rico 00732-7004, USA.
J Clin Immunol. 2001 Mar;21(2):135-44. doi: 10.1023/a:1011055805869.
Flow cytometric analysis of T cells from HIV+ and normal individuals activated for 15 hr showed that the percentage of cells producing interferon-gamma (INFgamma) was enhanced approximately threefold (39 compared to 14%) in the HIV+ CD8+ population. Activation modes, other than anti-CD3 with PMA, were ineffective, and in no case did the percentage of HIV+ CD4+ T cells show increased INFgamma production over controls. Enhanced INFgamma production was not induced by either anti-CD3 or PMA alone, or anti-CD3 or ConA with anti-CD28, or enhanced by N-acetylcysteine. In contrast to INFgamma production, the percentage of CD4+ T cells producing interleukin-2 (Il-2) greatly exceeded that of the CD8+ T cells. The results from flow cytometry analyses of HIV+ CD8+ T cells was supported by quantitative analysis of INFgamma mRNA (by PCR) and INFgamma secretion by ELISA. These methods showed a sixfold and three- to fivefold increase, respectively, on a per cell basis. As HIV infection progresses, as shown by loss of CD4+ T cells, the proportion of CD8+ CD28- T cells increases, and it is this T cell subset that is responsible for 80% or more of the enhanced INFgamma production. The enhanced INFgamma in HIV+ patients derives from two factors: the increase in CD8+ CD28- cells to 70% and the percentage producing INFgamma (60%, compared to 21% for CD8+ CD28+ cells). Our findings of a substantial increase in INFgamma production in HIV infection arising from the increased number of CD8+ CD28- T cells are compatible with clinical studies which show elevated INFgamma in HIV+ serum and INFgamma producing CD8+ T cells dominating HIV+ lymph nodes. We also found a significantly decreased proliferative response of the HIV+-derived CD8+ T cell fraction with coactivator anti-CD-28, in contrast to PMA (with anti-CD3), which is probably a reflection of the diminished population of CD8+ CD28+ T cells in HIV+ donors compared to normal donors (30.7 compared to 67.9%).
对来自HIV阳性个体和正常个体且已激活15小时的T细胞进行流式细胞术分析,结果显示,在HIV阳性CD8 +群体中,产生γ干扰素(INFγ)的细胞百分比提高了约三倍(从14%提高到39%)。除了用佛波酯(PMA)处理抗CD3外,其他激活方式均无效,并且在任何情况下,HIV阳性CD4 + T细胞产生INFγ的百分比均未超过对照组。单独的抗CD3或PMA,或抗CD3或伴刀豆球蛋白A(ConA)与抗CD28处理均未诱导INFγ产生增加,N - 乙酰半胱氨酸也未增强其产生。与INFγ产生情况相反,产生白细胞介素 - 2(Il - 2)的CD4 + T细胞百分比大大超过CD8 + T细胞。对HIV阳性CD8 + T细胞进行流式细胞术分析的结果得到了INFγ信使核糖核酸(mRNA,通过聚合酶链反应(PCR)测定)定量分析以及酶联免疫吸附测定(ELISA)检测INFγ分泌情况的支持。这些方法显示,以每个细胞为基础,INFγ mRNA增加了六倍,INFγ分泌增加了三到五倍。随着HIV感染的进展,如CD4 + T细胞数量减少所示,CD8 + CD28 - T细胞的比例增加,正是这个T细胞亚群导致了80%或更多的INFγ产生增加。HIV阳性患者中INFγ增加源于两个因素:CD8 + CD28 - 细胞增加到70%以及产生INFγ的百分比增加(60%,而CD8 + CD28 +细胞为21%)。我们发现,HIV感染中由于CD8 + CD28 - T细胞数量增加导致INFγ产生大幅增加,这与临床研究结果相符,临床研究显示HIV阳性血清中INFγ升高,且HIV阳性淋巴结中以产生INFγ的CD8 + T细胞为主。我们还发现,与PMA(与抗CD3联用)相比,共激活剂抗CD - 28处理使HIV来源的CD8 + T细胞部分的增殖反应显著降低,这可能反映了与正常供体相比,HIV阳性供体中CD8 + CD28 + T细胞数量减少(分别为30.7%和67.9%)。