Identification of T-cell epitopes in the structural and non-structural proteins of classical swine fever virus
Identification of T-cell epitopes in the structural and non-structural proteins of classical swine fever virus
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DOI：
10.1099/0022-1317-83-3-551
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发表时间：
2002-03-01
影响因子：
3.8
通讯作者：
Saalmüller, A
中科院分区：
文献类型：
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作者：
Armengol, E;Wiesmüller, KH;Saalmüller, A
为了鉴定猪瘟病毒（CSFV）新的t细胞表位，合成并筛选了573条重叠的合成五肽，覆盖了猪瘟病毒（Glentorf株）82%的基因组序列。在增殖试验中，分布在CSFV病毒蛋白序列中的26个多肽能够在感染CSFV- glentorf的d/d单倍型猪的PBMCs中诱导特异性t细胞反应。在这26个多肽中，18个也被CSFV-Alfort/187感染的d/d单倍型猪的pbmc识别。进一步实验表明，猪瘟病毒非结构蛋白NS2-3的1446 ~ 1460氨基酸残基对应的肽290 （KHKVRNEVMVHWFDD）在体外二次再刺激后可诱导干扰素γ分泌。这种五肽刺激t细胞的主要组织相容性复合体（MHC）限制被鉴定为主要是MHC II类和部分MHC i类。在细胞溶解实验中，猪瘟病毒特异性细胞毒性t淋巴细胞（ctl）能够裂解装载肽290的靶细胞。这些发现表明在该肽中存在csfv特异性辅助t细胞表位和CTL表位。
To identify new T-cell epitopes of classical swine fever virus (CSFV), 573 overlapping, synthetic pentadecapeptides spanning 82% of the CSFV (strain Glentorf) genome sequence were synthesized and screened. In proliferation assays, 26 peptides distributed throughout the CSFV viral protein sequences were able to induce specific T-cell responses in PBMCs from a CSFV-Glentorf-infected d/d haplotype pig. Of these 26 peptides, 18 were also recognized by PBMCs from a CSFV-Alfort/187-infected d/d haplotype pig. In further experiments, it could be shown that peptide 290 (KHKVRNEVMVHWFDD), which corresponds to amino acid residues 1446-1460 of the CSFV non-structural protein NS2-3 could induce interferon-gamma secretion after secondary in vitro restimulation. The major histocompatibility complex (MHC) restriction for stimulation of T-cells by this pentadecapeptide was identified as being mainly MHC class II and partially MHC class I. In cytolytic assays, CSFV-specific cytotoxic T-lymphocytes (CTLs) were able to lyse peptide 290-loaded target cells. These findings indicate the existence of a CSFV-specific helper T-cell epitope and a CTL epitope in this peptide.