Long-term culture and fine specificity of human cytotoxic T-lymphocyte clones reactive with human immunodeficiency virus type 1.
Long-term culture and fine specificity of human cytotoxic T-lymphocyte clones reactive with human immunodeficiency virus type 1.
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与 1 型人类免疫缺陷病毒反应的人类细胞毒性 T 淋巴细胞克隆的长期培养和良好特异性。
DOI：
10.1073/pnas.86.23.9514
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发表时间：
1989
影响因子：
11.1
通讯作者：
R. Schooley
中科院分区：
文献类型：
--
作者：
B. Walker;C. Flexner;K. Birch;Laura Fisher;T. Paradis;A. Aldovini;R. Young;B. Moss;R. Schooley
人类免疫缺陷病毒1型（HIV-1）免疫原性表位的定义对艾滋病疫苗策略的合理设计至关重要。在这项研究中，我们从两名血清阳性受试者的外周血中生成了7个HIV-1逆转录酶特异性细胞毒性t淋巴细胞（CTL）克隆。这些CTL克隆识别的表位是用表达截断逆转录酶蛋白的重组hiv -1痘苗病毒载体感染的靶细胞鉴定的，并通过用重叠的25个氨基酸合成逆转录酶肽孵育的靶细胞进一步确定。鉴定出5个不同的CTL表位，每个表位的识别都受到I类人白细胞抗原（HLA）的限制。克隆在连续培养中保持特异性的细胞溶解功能长达11个月，只需要定期用cd3特异性单克隆抗体进行再刺激。这些结果表明，hiv -1特异性，主要组织相容性i类限制性CTL识别单个病毒基因产物与不同宿主HLA抗原的多个表位。此外，他们还证明，人类病毒特异性CTL可以在长期培养中生长，而无需再次暴露于病毒抗原。
The definition of human immunodeficiency virus type 1 (HIV-1) immunogenic epitopes is central to the rational design of AIDS vaccine strategies. In this study, we have generated seven HIV-1 reverse transcriptase-specific cytotoxic T-lymphocyte (CTL) clones from the peripheral blood of two seropositive subjects. Epitopes recognized by these CTL clones were identified by using target cells infected with recombinant HIV-1-vaccinia virus vectors expressing truncated reverse transcriptase proteins and further defined by using target cells incubated with overlapping 25-amino acid synthetic reverse transcriptase peptides. Five different CTL epitopes were identified, and in each case recognition was restricted by class I human leukocyte antigens (HLA). Clones maintained specific cytolytic function in continuous culture for up to 11 months, requiring only periodic restimulation with a CD3-specific monoclonal antibody. These results indicate that HIV-1-specific, major histocompatibility class I-restricted CTL recognize multiple epitopes of a single viral gene product in conjunction with different host HLA antigens. In addition, they demonstrate that human virus-specific CTL can be grown in long-term culture without the need for reexposure to viral antigen.