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目的:鉴定汉滩病毒核衣壳蛋白(HTNVNP)C 端T细胞表位,为肾综合征出血热(HFRS)发病机制、疫苗研制及抗病毒免 疫反应研究奠定基础.方法:采用Ficoll密度梯度离心法分离HFRS恢复期患者外周血单个核细胞(PBMC).用IFN-γELISPO 实验和T细胞增殖实验,测试7例患者PBMC对23条NPC-端合成多肽的T细胞应答.结果:IFN-γELISPOT实验结果表明, 名供体(3、4)可分别检测到对51、70号2条多肽特异性T细胞应答.在供体3,70号肽特异性T细胞频率为45SFC/10E6PBMC 在供体4,51号肽特异性T细胞频率为82SFC/10E6PBMC。T细胞增殖实验与ELISPOT结果基本一致,但53号肽和64号肽 还可分别刺激供体1和供体4的T细胞增殖,而未能诱导IFN-γ分泌.结论:51号和70号多肽可能是NPC-端较强的T细胞 表位.
Objective: To identify the C-terminal T cell epitopes of Hantaan virus nucleocapsid protein (HTNVNP) and lay a foundation for the pathogenesis of HFRS, vaccine development and antiviral immune response.Methods: Ficoll density gradient centrifugation Peripheral blood mononuclear cells (PBMCs) were isolated from patients with HFRS convalescence. T-cell responses to 23 NPC-synthesized peptides in 7 patients were tested by IFN-γELISPO assay and T cell proliferation assay.Results IFN-γELISPOT assay The results showed that the donor (3, 4) could detect 2 polypeptide-specific T cell responses to 51 and 70 respectively.When donor 3,70 peptide-specific T cell frequency was 45SFC / 10E6PBMC in donor 4 , 51 peptide-specific T cell frequency of 82SFC / 10E6PBMC. T cell proliferation assay was consistent with ELISPOT results, but peptide 53 and peptide 64 also stimulated T cell proliferation of donor 1 and donor 4, respectively, but failed to induce IFN-γ secretion.Conclusion: Polypeptides may be NPC-terminal T cell epitopes.