Enumeration of latently infected CD4+ T cells from HIV-1-infected patients using an HIV-1 antigen ELISPOT assay
Enumeration of latently infected CD4+ T cells from HIV-1-infected patients using an HIV-1 antigen ELISPOT assay
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DOI：
10.1016/s1386-6532(03)00083-0
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发表时间：
2004-01-01
影响因子：
8.8
通讯作者：
Vendrell, JP
中科院分区：
文献类型：
--
作者：
Fondere, JM;Planas, JF;Vendrell, JP
背景资料：潜伏感染的携带有复制能力的HIV-1的静息CD 4（+）T细胞存在于初始的慢性感染个体以及正在接受高效抗逆转录病毒治疗（HAART）的个体中。这些细胞作为病毒复制重新激活的潜在来源，仍然是根除HIV-1的主要障碍。目的：酶联免疫斑点（ELISPOT）试验适用于在单细胞水平上检测和计数HIV-1抗原分泌细胞。我们应用该检测来计数潜伏的HIV-1感染的CD 4（+）T细胞。研究设计：在HIV-1感染的静息CD 4（+）T细胞体外模型中以及在18名HAART治疗患者和4名HIV-1感染的未治疗患者中评估了潜伏感染的CD 4（+）T细胞。将富集的CD 4（+）T细胞与或不与抗CD 3和CD 28 T细胞受体的抗体以及来自HIV-1血清阴性个体的经辐照的外周血单核细胞一起培养。在细胞培养期，用HIV-1抗原ELISPOT试验检测CD 4（+）T淋巴细胞。结果：在实验性HIV-1感染模型中，经多克隆活化后，共计数到5579 +/-4190个分泌HIV-1抗原的CD 4（+）T细胞。相比之下，从未刺激的T细胞中仅获得15 +/- 6个HIV-1免疫斑点。在所有受试患者中，检测到诱导的HIV-1抗原分泌细胞的频率为55 +/- 108/10（6）CD 4（+）T细胞。结论：由于每个免疫斑点代表一个HIV-1抗原分泌细胞，因此HIV-1 ELISPOT检测试剂盒可有效计数潜伏感染HIV-1的循环CD 4（+）T淋巴细胞。(C)2003 Elsevier B. V.保留所有权利。
Background: Latently infected resting CD4(+) T cells carrying replication-competent HIV-1 are present in naive, chronically infected individuals as well as in those who are receiving highly active antiretroviral therapy (HAART). These cells serve as a potential source of reactivation of viral replication and remain a major obstacle for the eradication of HIV-1. Objectives: The enzyme-linked immunospot (ELISPOT) assay was adapted to the detection and the enumeration of HIV-1 antigen-secreting cells at the single cell level. We applied this test to count latently HIV-1-infected CD4(+) T cells. Study design: Latently infected CD4(+) T cells were assessed in an in vitro model of HIV-1-infected resting CD4(+) T cells as well as in eighteen HAART-treated and in four HIV-1-infected untreated patients. Enriched CD4(+) T cells were cultured with or without antibodies against CD3 and CD28 T cell receptors and with irradiated peripheral blood mononuclear cells from HIV-1 seronegative individuals. At the term of the cell culture, CD4(+) T lymphocytes were tested using the HIV-1 antigen ELISPOT assay. Results: In the experimental HIV-1 infection model, 5579 +/- 4190 CD4(+) T cells secreting HIV-1 antigen were enumerated after polyclonal activation. In contrast, only 15 +/- 6 HIV-1 immunospots were obtained from unstimulated T cells. In all patients tested, induced HIV-1 antigen-secreting cells were measured at a frequency of 55 +/- 108/10(6) CD4(+) T cells. Conclusion: As each immunospot represents one HIV-1 antigen-secreting cell, the HIV-1 ELISPOT assay is a powerful to enumerate circulating CD4(+) T lymphocytes latently infected with HIV-1. (C) 2003 Elsevier B.V. All rights reserved.