鉴定登革热病毒特异性 HLA-A*0201 限制性 CD8+ T 细胞表位。

Identification of a dengue virus-specific HLA-A*0201-restricted CD8+ T cell epitope.

机构信息

Department of Microbiology, Zhongshan Medical School, Sun Yat-Sen University, Guangzhou, China.

出版信息

J Med Virol. 2010 Apr;82(4):642-8. doi: 10.1002/jmv.21736.

PMID:20166182

Abstract

In this study, a combination of epitope-prediction programs and in vitro assays was used to identify dengue virus (DENV)-specific CD8(+) T cell epitopes. Peripheral blood mononuclear cells (PBMCs) isolated from patients who recovered from dengue fever were stimulated with candidate epitope peptides derived from DENV, which were predicted by using SYFPEITHI and RANKpep epitope-prediction programs. The IFN-gamma ELISpot results and the results of intracellular staining of IFN-gamma showed that peptides NS4b_40 (TLYAVATTI), E_256 (QEGAMHTAL), NS3_205 (LPAIVREAI), NS5_210 (SRNSTHEMY), and NS3_207 (AIVREAIKR) could induce the recall response of CD8(+) T cells. Furthermore, the results of the MHC-peptide complex stabilization assay revealed that peptide NS4b_40 (TLYAVATTI) has a high affinity for HLA-A0201 molecules. The IFN-gamma ELISpot results and staining of intracellular IFN-gamma confirmed that this peptide could induce high-level CD8(+) T cell response in HLA-A0201 positive PBMCs. Peptide NS4b_40 (TLYAVATTI) was identified as a novel DENV-specific HLA-A*0201-restricted CD8(+) T cell epitope.

摘要

在这项研究中，采用了表位预测程序和体外测定法来鉴定登革热病毒（DENV）特异性 CD8+ T 细胞表位。从登革热康复患者中分离出外周血单核细胞（PBMC），用 SYFPEITHI 和 RANKpep 表位预测程序预测的 DENV 衍生候选表位肽刺激这些 PBMC。IFN-γ ELISpot 结果和 IFN-γ 细胞内染色结果表明，肽 NS4b_40（TLYAVATTI）、E_256（QEGAMHTAL）、NS3_205（LPAIVREAI）、NS5_210（SRNSTHEMY）和 NS3_207（AIVREAIKR）能够诱导 CD8+T 细胞的回忆反应。此外，MHC-肽复合物稳定测定的结果表明，肽 NS4b_40（TLYAVATTI）与 HLA-A0201 分子具有高亲和力。IFN-γ ELISpot 结果和 IFN-γ 细胞内染色证实，这种肽能够在 HLA-A0201 阳性 PBMC 中诱导高水平的 CD8+T 细胞反应。肽 NS4b_40（TLYAVATTI）被鉴定为一种新的 DENV 特异性 HLA-A*0201 限制性 CD8+T 细胞表位。