Draenert R, Verrill C L, Tang Y, Allen T M, Wurcel A G, Boczanowski M, Lechner A, Kim A Y, Suscovich T, Brown N V, Addo M M, Walker B D
Howard Hughes Medical Institute, Partners AIDS Research Center, Massachusetts General Hospital and Harvard Medical School Division of AIDS, Boston, Massachusetts 02129, USA.
J Virol. 2004 Jan;78(2):630-41. doi: 10.1128/jvi.78.2.630-641.2004.
CD8 T-cell responses are thought to be crucial for control of viremia in human immunodeficiency virus (HIV) infection but ultimately fail to control viremia in most infected persons. Studies in acute infection have demonstrated strong CD8-mediated selection pressure and evolution of mutations conferring escape from recognition, but the ability of CD8 T-cell responses that persist in late-stage infection to recognize viruses present in vivo has not been determined. Therefore, we studied 24 subjects with advanced HIV disease (median viral load = 142,000 copies/ml; median CD4 count = 71/ micro l) and determined HIV-1-specific CD8 T-cell responses to all expressed viral proteins using overlapping peptides by gamma interferon Elispot assay. Chronic-stage virus was sequenced to evaluate autologous sequences within Gag epitopes, and functional avidity of detected responses was determined. In these subjects, the median number of epitopic regions targeted was 13 (range, 2 to 39) and the median cumulative magnitude of CD8 T-cell responses was 5,760 spot-forming cells/10(6) peripheral blood mononuclear cells (range, 185 to 24,700). On average six (range, one to 8) proteins were targeted. For 89% of evaluated CD8 T-cell responses, the autologous viral sequence was predicted to be well recognized by these responses and the majority of analyzed optimal epitopes were recognized with medium to high functional avidity by the contemporary CD8 T cells. Withdrawal of antigen by highly active antiretroviral therapy led to a significant decline both in breadth (P = 0.032) and magnitude (P = 0.0098) of these CD8 T-cell responses, providing further evidence that these responses had been driven by recognition of autologous virus. These results indicate that strong, broadly directed, and high-avidity gamma-interferon-positive CD8 T-cells directed at autologous virus persist in late disease stages, and the absence of mutations within viral epitopes indicates a lack of strong selection pressure mediated by these responses. These data imply functional impairment of CD8 T-cell responses in late-stage infection that may not be reflected by gamma interferon-based screening techniques.
CD8 T细胞反应被认为对控制人类免疫缺陷病毒(HIV)感染中的病毒血症至关重要,但在大多数感染者中最终无法控制病毒血症。急性感染研究表明,CD8介导的选择压力很强,且会发生导致病毒逃避识别的突变演变,但尚未确定晚期感染中持续存在的CD8 T细胞反应识别体内存在的病毒的能力。因此,我们研究了24名晚期HIV疾病患者(病毒载量中位数=142,000拷贝/毫升;CD4计数中位数=71/微升),并通过γ干扰素酶联免疫斑点试验,使用重叠肽确定了HIV-1特异性CD8 T细胞对所有表达的病毒蛋白的反应。对慢性期病毒进行测序,以评估Gag表位内的自体序列,并确定检测到的反应的功能亲和力。在这些受试者中,靶向的表位区域中位数为13个(范围为2至39个),CD8 T细胞反应的累计幅度中位数为5,760个斑点形成细胞/10⁶外周血单个核细胞(范围为185至24,700)。平均有6种(范围为1至8种)蛋白质被靶向。对于89%的评估CD8 T细胞反应,预测自体病毒序列能被这些反应很好地识别,并且大多数分析的最佳表位被当代CD8 T细胞以中等到高的功能亲和力识别。通过高效抗逆转录病毒疗法去除抗原导致这些CD8 T细胞反应的广度(P=0.032)和幅度(P=0.0098)均显著下降,进一步证明这些反应是由对自体病毒的识别驱动的。这些结果表明,针对自体病毒的强大、广泛定向且高亲和力的γ干扰素阳性CD8 T细胞在疾病晚期持续存在,并且病毒表位内没有突变表明缺乏由这些反应介导的强大选择压力。这些数据意味着晚期感染中CD8 T细胞反应存在功能损害,而基于γ干扰素的筛选技术可能无法反映这一点。