Zivny J, Kurane I, Leporati A M, Ibe M, Takiguchi M, Zeng L L, Brinton M A, Ennis F A
Department of Medicine, University of Massachusetts Medical Center, Worcester 01655, USA.
J Exp Med. 1995 Sep 1;182(3):853-63. doi: 10.1084/jem.182.3.853.
It is generally accepted that virus-specific CD8+ cytotoxic T lymphocytes (CTLs) recognize nine-amino acid peptides in conjunction with HLA class I molecules. We recently reported that dengue virus-specific CD8+ CTLs of two different serotype specificities, which were established by stimulation with dengue virus, recognize a single nine-amino acid peptide of the nonstructural protein NS3 of dengue virus type 4 (D4V) in an HLA-B35-restricted fashion. To further analyze the relationships between the serotype specificities of T cells and the amino acid sequence of the recognized peptides, we examined the ability of this viral peptide D4.NS3.500-508 (TPEGIIPTL) to stimulate T lymphocytes of an HLA-B35-positive, dengue virus type 4-immune donor. Peptide stimulation of the PBMC generated dengue virus-specific, HLA-B-35-restricted CD8+ CTL clones. These clones lysed dengue virus-infected autologous cells, as well as autologous target cells pulsed with this peptide. Four patterns of dengue virus serotype specificities were demonstrated on target cells infected with dengue-vaccinia recombinant viruses or pulsed with synthetic peptides corresponding to amino acid sequences of four dengue virus serotypes. Two serotype-specific clones recognized only D4V. Three dengue virus subcomplex-specific clones recognized D1V, D3V, and D4V, and one subcomplex-specific clone recognized D2V and D4V. Three dengue virus serotype-cross-reactive clones recognized D1V-D4V. Thus, a single nine-amino acid peptide induces proliferation of a heterogeneous panel of dengue virus-specific CD8+ CTL clones that are all restricted by HLA-B35 but have a variety of serotype specificities. Peptides that contain a single amino acid substitution at each position of D4.NS3.500-508 were recognized differently by the T cell clones. These results indicate that a single epitope can be recognized by multiple CD8+ CTLs that have a variety of serotype specificities, but the manner of recognition by these multiple CTLs is heterogeneous.
普遍认为，病毒特异性CD8 + 细胞毒性T淋巴细胞（CTL）与HLA I类分子结合识别九氨基酸肽。我们最近报道，通过登革病毒刺激建立的两种不同血清型特异性的登革病毒特异性CD8 + CTL，以HLA - B35限制的方式识别登革病毒4型（D4V）非结构蛋白NS3的单个九氨基酸肽。为了进一步分析T细胞血清型特异性与识别肽氨基酸序列之间的关系，我们检测了这种病毒肽D4.NS3.500 - 508（TPEGIIPTL）刺激HLA - B35阳性、登革病毒4型免疫供体的T淋巴细胞的能力。肽刺激外周血单核细胞（PBMC）产生了登革病毒特异性、HLA - B - 35限制的CD8 + CTL克隆。这些克隆裂解登革病毒感染的自体细胞以及用该肽脉冲处理的自体靶细胞。在用登革痘苗重组病毒感染或用对应于四种登革病毒血清型氨基酸序列的合成肽脉冲处理的靶细胞上，证明了四种登革病毒血清型特异性模式。两个血清型特异性克隆仅识别D4V。三个登革病毒亚复合体特异性克隆识别D1V、D3V和D4V，一个亚复合体特异性克隆识别D2V和D4V。三个登革病毒血清型交叉反应性克隆识别D1V - D4V。因此，单个九氨基酸肽诱导了一组异质性的登革病毒特异性CD8 + CTL克隆的增殖，这些克隆均受HLA - B35限制，但具有多种血清型特异性。在D4.NS3.500 - 508的每个位置含有单个氨基酸替代的肽被T细胞克隆以不同方式识别。这些结果表明，单个表位可被具有多种血清型特异性的多个CD8 + CTL识别，但这些多个CTL的识别方式是异质性的。
