Janossy G, Borthwick N, Lomnitzer R, Medina E, Squire S B, Phillips A N, Lipman M, Johnson M A, Lee C, Bofill M
Department of Clinical Immunology, Royal Free Hospital and School of Medicine, London, UK.
AIDS. 1993 May;7(5):613-24. doi: 10.1097/00002030-199305000-00002.
The proliferative defects of CD4 and CD8 cells taken from 474 HIV-1-seropositive individuals during various stages of disease were quantitated. Phytohaemagglutinin (PHA) and soluble anti-CD3 were used in optimal mitogenic concentrations in the presence of recombinant interleukin-2 (rIL-2) and conditioned medium, and the proliferation of cells from HIV-1-seropositive donors was assessed in co-culture with HIV-1-seronegative cells in order to exclude effects of cytokine deficiency. Defects within the CD45RA+ ('unprimed') and CD45R0+ ('primed') T-cell populations were also investigated.
Quantitative immunofluorescence and double and triple labelling in flow cytometry were performed for (1) CD25 (IL-2 receptor alpha chain) expression, (2) lymphocyte and T-cell survival, and (3) blast transformation and proliferation--in relation to the original input of cells for each subpopulation.
T cells from normal and HIV-1-seropositive donors were CD25+ at day 1. In HIV-1-seropositive patients a variable number of CD4 and CD8 lymphocytes failed to further increase CD25, and died as a sign of activation-associated lymphocyte death (AALD). Forty-two per cent of asymptomatic subjects, including 32% of those with CD4 cell counts > 400 x 10(6)/l, showed a poor blast transformation (< 30% blasts). Cells from HIV-1-seropositive donors showed poor blast responses when co-cultured with HIV-1-seronegative cells; both CD4 and CD8 cells were handicapped. In asymptomatic HIV-1-seropositive people T cells with the CD45R0+ RA- ('primed') phenotype were three to five times more vulnerable to AALD than the CD45RA+ RO- ('unprimed') cells. In patients in Centers for Disease Control and Prevention (CDC) disease stage IV both CD45R0+ and -RA+ populations were severely affected.
This is the first quantitative analysis to demonstrate that in HIV-1 infection mitogen-stimulated CD45R0+ ('primed') T cells preferentially die upon activation. Both the CD4 and CD8 lineages are affected, as seen in animal models of graft versus host disease. AALD may explain defects of immunological memory. The analysis of AALD may be a suitable assay for studying whether antiviral drugs influence the proliferative responses of lymphocytes.
对474例处于疾病不同阶段的HIV-1血清阳性个体的CD4和CD8细胞的增殖缺陷进行定量分析。在重组白细胞介素-2（rIL-2）和条件培养基存在的情况下，使用最佳促有丝分裂浓度的植物血凝素（PHA）和可溶性抗CD3，并将HIV-1血清阳性供体的细胞与HIV-1血清阴性细胞共培养，以评估细胞增殖，从而排除细胞因子缺乏的影响。还对CD45RA+（“未致敏”）和CD45R0+（“致敏”）T细胞群体中的缺陷进行了研究。
采用定量免疫荧光以及流式细胞术中的双重和三重标记法，对以下方面进行检测：（1）CD25（IL-2受体α链）表达；（2）淋巴细胞和T细胞存活情况；（3）与每个亚群细胞的原始输入量相关的母细胞转化和增殖情况。
正常和HIV-1血清阳性供体的T细胞在第1天时为CD25+。在HIV-1血清阳性患者中，数量不等的CD4和CD8淋巴细胞未能进一步增加CD25表达，并发生死亡，这是活化相关淋巴细胞死亡（AALD）的表现。42%的无症状受试者，包括32% CD4细胞计数>400×10⁶/L的受试者，表现出较差的母细胞转化（<30%母细胞）。HIV-1血清阳性供体的细胞与HIV-1血清阴性细胞共培养时，母细胞反应较差；CD4和CD8细胞均存在缺陷。在无症状的HIV-1血清阳性个体中，具有CD45R0+ RA-（“致敏”）表型的T细胞比CD45RA+ RO-（“未致敏”）细胞更容易发生AALD，前者的易感性是后者的三到五倍。在疾病控制和预防中心（CDC）IV期的患者中，CD45R0+和-RA+群体均受到严重影响。
这是首次进行的定量分析，表明在HIV-1感染中，有丝分裂原刺激的CD45R0+（“致敏”）T细胞在活化时优先死亡。CD4和CD8谱系均受到影响，这与移植物抗宿主病动物模型中的情况一致。AALD可能解释免疫记忆缺陷。对AALD的分析可能是一种合适的检测方法，用于研究抗病毒药物是否影响淋巴细胞的增殖反应。
