Holzer T J, Heynen C A, Novak R M, Pitrak D L, Dawson G J
Abbott Laboratories, Diagnostics Division, Abbott Park, IL 60064-3500.
AIDS. 1993 Nov;7 Suppl 2:S3-5. doi: 10.1097/00002030-199311002-00002.
Markers of HIV disease progression such as soluble p24 antigen detection and CD4 lymphocyte depletion are most useful in the later stages of HIV disease and are relatively insensitive as therapeutic monitors. Flow cytometric detection of HIV-1 replication in CD4 lymphocytes was evaluated for use as a marker in predicting disease progression earlier in the course of HIV disease.
To determine whether the number of HIV-1-infected CD4 cells, as measured by p24 antigen detection, can be correlated with disease progression, we used flow cytometry to detect intracellular HIV-1 p24 in CD4 lymphocytes from HIV-1-seropositive subjects at all stages of HIV disease.
Mononuclear cells from HIV-1-seropositive subjects and uninfected control subjects were permeabilized and stained with anti-HIV-1 p24 monoclonal antibodies. The cells were then stained with a fluorescein isothiocyanate-conjugated goat antimurine immunoglobulin G followed by a phycoerythrin-conjugated monoclonal anti-CD4 antibody. The percentage of p24-positive CD4 lymphocytes was compared with absolute CD4 counts, soluble p24 detection and Walter Reed classification.
CD4 lymphocyte absolute counts and the percentage of CD4 lymphocytes declined as the Walter Reed classification indicated disease progression. The mean percentage of p24 antigen-positive CD4 lymphocytes increased with disease progression. Only 30% of Walter Reed stage 6 subjects were soluble p24 antigen-positive, whereas 68% were cellular p24 antigen-positive.
The percentage of p24 antigen-positive CD4 lymphocytes increased as HIV disease progressed. Flow cytometric quantitation of p24 antigen-positive CD4 cells is a useful method of monitoring in vivo HIV replication and disease progression.
诸如可溶性p24抗原检测和CD4淋巴细胞耗竭等HIV疾病进展标志物在HIV疾病后期最为有用，作为治疗监测指标相对不敏感。评估流式细胞术检测CD4淋巴细胞中HIV-1复制情况，以作为在HIV疾病进程中更早预测疾病进展的标志物。
为确定通过p24抗原检测测得的HIV-1感染的CD4细胞数量是否与疾病进展相关，我们使用流式细胞术检测HIV疾病各阶段HIV-1血清阳性受试者CD4淋巴细胞内的HIV-1 p24。
对HIV-1血清阳性受试者和未感染对照受试者的单核细胞进行通透处理，并用抗HIV-1 p24单克隆抗体染色。然后用异硫氰酸荧光素偶联的山羊抗鼠免疫球蛋白G染色，接着用藻红蛋白偶联的抗CD4单克隆抗体染色。将p24阳性CD4淋巴细胞的百分比与CD4绝对计数、可溶性p24检测及沃尔特·里德分类法进行比较。
随着沃尔特·里德分类法表明疾病进展，CD4淋巴细胞绝对计数和CD4淋巴细胞百分比下降。p24抗原阳性CD4淋巴细胞的平均百分比随疾病进展而增加。沃尔特·里德6期受试者中只有30%的人可溶性p24抗原呈阳性，而68%的人细胞p24抗原呈阳性。
随着HIV疾病进展，p24抗原阳性CD4淋巴细胞的百分比增加。流式细胞术定量检测p24抗原阳性CD4细胞是监测体内HIV复制和疾病进展的一种有用方法。
