Lederman M M, Purvis S F, Walter E I, Carey J T, Medof M E
Department of Medicine, Case Western Reserve University, Cleveland, OH 44106.
Proc Natl Acad Sci U S A. 1989 Jun;86(11):4205-9. doi: 10.1073/pnas.86.11.4205.
Although the human immunodeficiency virus can induce cytopathic changes in human lymphocytes in vitro, the mechanism(s) underlying progressive lymphopenia in patients with AIDS and AIDS-related complex has not been elucidated. To investigate this issue, peripheral blood lymphocytes of AIDS and AIDS-related complex patients and healthy control subjects were examined for their ability to resist homologous complement-mediated lysis. Upon sensitization with monoclonal antibodies to major histocompatibility complex class I antigen, as much as 48% lysis of patients' cells was observed in as little as a 1:32 dilution of human serum compared to 18 +/- 8% (mean +/- SD) lysis of controls' cells even in a 1:8 dilution of human serum. To investigate the mechanism of the abnormal complement sensitivity, AIDS and AIDS-related complex cells were analyzed for expression of decay-accelerating factor (DAF), a complement regulatory protein that functions intrinsically in blood cell membranes to prevent complement activation on their surfaces. Flow cytometric assays using anti-DAF monoclonal antibodies demonstrated that patients' lymphocytes and monocytes were DAF-deficient, in contrast to their polymorphonuclear leukocytes, which showed normal DAF levels. Expression of DAF was diminished on CD4+ as well as CD8+ T-lymphocyte subpopulations as opposed to expression of CD3, which was comparable in patients and controls. Incubation of normal lymphocytes with anti-DAF monoclonal antibodies or phosphatidylinositol-specific phospholipase C, an enzyme that cleaves DAF, enhanced lysis. Conversely, reconstitution of patients' cells with exogenous DAF reduced their lysis. The findings of heightened complement sensitivity and DAF deficiency of patients' lymphocytes in vitro suggest the possibility that the DAF deficit may contribute to the progressive lymphopenia of AIDS in vivo.
虽然人类免疫缺陷病毒在体外可诱导人淋巴细胞发生细胞病变,但艾滋病患者和艾滋病相关综合征患者出现进行性淋巴细胞减少的潜在机制尚未阐明。为研究此问题,对艾滋病患者、艾滋病相关综合征患者及健康对照者的外周血淋巴细胞抵抗同源补体介导裂解的能力进行了检测。用针对主要组织相容性复合体I类抗原的单克隆抗体致敏后,在人血清稀释至1:32时,就观察到患者细胞高达48%的裂解,而对照者细胞即使在人血清稀释至1:8时,裂解率也仅为18±8%(平均值±标准差)。为研究补体敏感性异常的机制,对艾滋病患者和艾滋病相关综合征患者的细胞进行了衰变加速因子(DAF)表达分析,DAF是一种补体调节蛋白,在血细胞细胞膜内发挥作用,防止补体在其表面激活。使用抗DAF单克隆抗体的流式细胞术检测表明,患者的淋巴细胞和单核细胞缺乏DAF,而其多形核白细胞DAF水平正常。与CD3表达在患者和对照者中相当不同,DAF在CD4⁺以及CD8⁺T淋巴细胞亚群上的表达减少。用抗DAF单克隆抗体或磷脂酰肌醇特异性磷脂酶C(一种可裂解DAF的酶)孵育正常淋巴细胞可增强裂解。相反,用外源性DAF重建患者细胞可降低其裂解率。体外患者淋巴细胞补体敏感性增强和DAF缺乏的研究结果提示,DAF缺陷可能在体内导致艾滋病患者进行性淋巴细胞减少。