Shankar P, Russo M, Harnisch B, Patterson M, Skolnik P, Lieberman J
Center for Blood Research, Harvard Medical School, and the New England Medical Center, Tufts University School of Medicine, Boston, MA, USA.
Blood. 2000 Nov 1;96(9):3094-101.
The functional status of circulating human immunodeficiency (HIV)-specific CD8 T cells in chronically infected subjects was evaluated. By flow cytometry, only 5 of 7 subjects had detectable CD8 T cells that produced IFN-gamma after stimulation with HIV-infected primary CD4 T cells. In 2 subjects, the frequency of IFN-gamma-producing cells increased 4-fold when IL-2 was added to the culture medium; in another subject, IFN-gamma-producing cells could be detected only after IL-2 was added. IFN-gamma-producing cells ranged from 0.4% to 3% of CD8 T cells. Major histocompatibility complex-peptide tetramer staining, which identifies antigen-specific T cells irrespective of function, was used to evaluate the proportion of HIV-specific CD8 T cells that may be nonfunctional in vivo. CD8 T cells binding to tetramers complexed to HIV gag epitope SLYNTVATL and reverse transcriptase epitope YTAFTIPSI were identified in 9 of 15 and 5 of 12 HLA-A2-expressing seropositive subjects at frequencies of 0.1% to 1.1% and 0.1 to 0.7%, respectively. Freshly isolated tetramer-positive cells expressed a mixed pattern of memory and effector markers. On average, IFN-gamma was produced by less than 25% of tetramer-positive CD8 T cells after stimulation with the relevant gag or reverse transcriptase peptide. In all subjects tested, freshly isolated CD8 T cells were not cytolytic against peptide-pulsed B lymphoblastoid cell line or primary HIV-infected CD4 T-cell targets. Exposure to IL-2 enhanced the cytotoxicity of CD8 T cells against primary HIV-infected CD4 targets in 2 of 2 subjects tested. These results suggest that a significant proportion of HIV-specific CD8 T cells may be functionally compromised in vivo and that some function can be restored by exposure to IL-2.
对慢性感染受试者体内循环的人免疫缺陷病毒(HIV)特异性CD8 T细胞的功能状态进行了评估。通过流式细胞术,7名受试者中只有5名的CD8 T细胞在受到HIV感染的原代CD4 T细胞刺激后可检测到产生γ干扰素。在2名受试者中,当向培养基中添加白细胞介素-2(IL-2)时,产生γ干扰素的细胞频率增加了4倍;在另一名受试者中,只有在添加IL-2后才能检测到产生γ干扰素的细胞。产生γ干扰素的细胞占CD8 T细胞的0.4%至3%。主要组织相容性复合体肽四聚体染色可识别抗原特异性T细胞而不考虑其功能,用于评估体内可能无功能的HIV特异性CD8 T细胞的比例。在15名表达HLA-A2的血清阳性受试者中的9名以及12名中的5名中,分别鉴定出与HIV gag表位SLYNTVATL和逆转录酶表位YTAFTIPSI复合的四聚体结合的CD8 T细胞,频率分别为0.1%至1.1%和0.1%至0.7%。新鲜分离的四聚体阳性细胞表达记忆和效应标志物的混合模式。平均而言,在用相关的gag或逆转录酶肽刺激后,不到25%的四聚体阳性CD8 T细胞产生γ干扰素。在所有测试的受试者中,新鲜分离的CD8 T细胞对肽脉冲的B淋巴母细胞系或原代HIV感染的CD4 T细胞靶标无细胞毒性。在2名测试的受试者中,暴露于IL-2增强了CD8 T细胞对原代HIV感染的CD4靶标的细胞毒性。这些结果表明,相当一部分HIV特异性CD8 T细胞在体内可能功能受损,并且通过暴露于IL-2可以恢复一些功能。