Littaua R A, Oldstone M B, Takeda A, Debouck C, Wong J T, Tuazon C U, Moss B, Kievits F, Ennis F A
Department of Medicine, University of Massachusetts Medical Center, Worcester 01655.
J Virol. 1991 Aug;65(8):4051-6. doi: 10.1128/JVI.65.8.4051-4056.1991.
A unique epitope on the gag protein of human immunodeficiency virus type 1 (HIV-1), located at amino acid 145 to 150, has been mapped by using a CD8+ cytotoxic T-lymphocyte (CTL) clone. This epitope is highly conserved among 18 HIV-1 strains. The HIV-1 gag-specific human leukocyte antigen (HLA) class I-restricted CD8+ CTL clone was generated from fresh peripheral blood mononuclear cells of an HIV-seropositive donor by stimulation with gamma-irradiated allogeneic peripheral blood mononuclear cells in the presence of an anti-CD3 monoclonal antibody and recombinant interleukin-2. This gag-specific CTL clone killed autologous target cells infected with a recombinant vaccinia virus containing the gag gene of HIV-1 and target cells pulsed with an authentic p24gag construct expressed in Escherichia coli. Fine specificity was determined by using a panel of overlapping 30-amino-acid-long synthetic peptides and subsequently using smaller peptides to precisely map the CTL domain on p24. The epitope is on a highly conserved region, and it overlaps with a major B-cell epitope of gag. This CD8+ T-cell epitope is restricted by HLA-Cw3, which has not been previously identified as a restricting element for human CTL responses.
利用一个CD8 + 细胞毒性T淋巴细胞（CTL）克隆，已绘制出1型人类免疫缺陷病毒（HIV - 1）gag蛋白上一个位于氨基酸145至150处的独特表位。该表位在18株HIV - 1毒株中高度保守。HIV - 1 gag特异性人类白细胞抗原（HLA）I类限制性CD8 + CTL克隆是通过在抗CD3单克隆抗体和重组白细胞介素 - 2存在的情况下，用γ射线照射的同种异体外周血单核细胞刺激一名HIV血清阳性供体的新鲜外周血单核细胞而产生的。这个gag特异性CTL克隆杀死了被含有HIV - 1 gag基因的重组痘苗病毒感染的自体靶细胞，以及用在大肠杆菌中表达的真实p24gag构建体脉冲处理过的靶细胞。通过使用一组重叠的30个氨基酸长的合成肽，随后使用更小的肽来精确绘制p24上的CTL结构域，确定了精细特异性。该表位位于一个高度保守的区域，并且与gag的一个主要B细胞表位重叠。这个CD8 + T细胞表位受HLA - Cw3限制，HLA - Cw3以前尚未被鉴定为人类CTL反应的限制元件。
