HIV-specific CD8(+) T cells produce antiviral cytokines but are impaired in cytolytic function.
HIV-specific CD8(+) T cells produce antiviral cytokines but are impaired in cytolytic function.
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DOI：
10.1084/jem.192.1.63
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发表时间：
2000-07-03
影响因子：
15.3
通讯作者：
Rowland-Jones, S L
中科院分区：
文献类型：
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作者：
Appay, V;Nixon, D F;Donahoe, S M;Gillespie, G M;Dong, T;King, A;Ogg, G S;Spiegel, H M;Conlon, C;Spina, C A;Havlir, D V;Richman, D D;Waters, A;Easterbrook, P;McMichael, A J;Rowland-Jones, S L
利用肽-人组织相容性白细胞抗原（HLA） I类四聚体复合物来鉴定抗原特异性CD8+ T细胞，为我们理解它们在控制病毒感染中的作用提供了重大进展。然而，这些细胞的确切功能仍然存在疑问，特别是在HIV感染中。病毒特异性的细胞毒性T淋巴细胞通过分泌可溶性因子如细胞因子和趋化因子来发挥其活性。我们在这里描述了一种方法，结合使用四聚体和细胞内染色来检查抗原特异性CD8+ T细胞的功能异质性。经特异性肽抗原刺激后，用FACS®分析四聚体阳性人群外周血中干扰素(IFN)-γ、肿瘤坏死因子(TNF)-α、巨噬细胞炎症蛋白(MIP)-1β和穿孔素的分泌情况。使用这种方法，我们评估了HIV特异性CD8+ T细胞与巨细胞病毒（CMV）特异性CD8+ T细胞在HIV慢性感染中的功能表型。我们发现，大多数针对CMV和HIV抗原特异性的循环CD8+ T细胞在应答抗原时，在分泌抗病毒细胞因子方面具有功能活性，尽管鉴定出一部分四聚体染色细胞分泌IFN-γ和MIP-1β，但不分泌TNF-α。然而，一个惊人的发现是hiv特异性CD8+ T细胞表达的穿孔素水平明显低于cmv特异性CD8+ T细胞。穿孔素的缺乏与hiv特异性细胞上CD27的持续表达有关，表明成熟受损，与来自同一供体的cmv特异性细胞相比，hiv特异性细胞的体外特异性裂解更低。因此，hiv特异性CD8+ T细胞的细胞溶解活性受损。
The use of peptide–human histocompatibility leukocyte antigen (HLA) class I tetrameric complexes to identify antigen-specific CD8+ T cells has provided a major development in our understanding of their role in controlling viral infections. However, questions remain about the exact function of these cells, particularly in HIV infection. Virus-specific cytotoxic T lymphocytes exert much of their activity by secreting soluble factors such as cytokines and chemokines. We describe here a method that combines the use of tetramers and intracellular staining to examine the functional heterogeneity of antigen-specific CD8+ T cells ex vivo. After stimulation by specific peptide antigen, secretion of interferon (IFN)-γ, tumor necrosis factor (TNF)-α, macrophage inflammatory protein (MIP)-1β, and perforin is analyzed by FACS® within the tetramer-positive population in peripheral blood. Using this method, we have assessed the functional phenotype of HIV-specific CD8+ T cells compared with cytomegalovirus (CMV)-specific CD8+ T cells in HIV chronic infection. We show that the majority of circulating CD8+ T cells specific for CMV and HIV antigens are functionally active with regards to the secretion of antiviral cytokines in response to antigen, although a subset of tetramer-staining cells was identified that secretes IFN-γ and MIP-1β but not TNF-α. However, a striking finding is that HIV-specific CD8+ T cells express significantly lower levels of perforin than CMV-specific CD8+ T cells. This lack of perforin is linked with persistent CD27 expression on HIV-specific cells, suggesting impaired maturation, and specific lysis ex vivo is lower for HIV-specific compared with CMV-specific cells from the same donor. Thus, HIV-specific CD8+ T cells are impaired in cytolytic activity.