摘要
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)发病机理、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 γELISPOT实验和T细胞增殖实验 ,测试 7名患者PBMC对 2 3条NPC 端合成多肽的T细胞应答。结果 IFN γELISPOT实验结果表明 ,2名供体(3、4 )可分别检测到对 5 1、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4 5SFC 10 6 PBMC ;在供体 4 ,5 1号肽特异性T细胞频率为 82SFC 10 6 PBMC。T细胞增殖实验与ELISPOT结果基本一致 ,但 5 3号肽和 6 4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 5 1号和 70号多肽可能是NPC 端较强的T细胞表位。
Objective To identify T cell epitopes on hantaan virus nucleocapsid protein, to lay the foundation for an understanding of HFRS pathogenesis and the design of vaccine strategies. Methods Peripheral blood mononuclear cells(PBMCs) were separated by Ficoll-hypaque density gradient centrifugation from HFRS patients. T cell response to 23 polypeptides synthesized based on nucleocapsid protein was examined by IFN-γ ELISPOT and T cell proliferation assay in 7 patients with hemorrhagic fever with renal syndrome (HFRS) in the convalescent phase. Results IFN-γ ELISPOT analysis showed that of the 7 individuals tested, two had detectable HTNV-specfic T cell responses to NP peptides. The frequency of No.70 and No.51 peptide-specific T cell was 45 and 82 SFC/10 6PBMCs in the donor 3 and 4 respectively. The results obtained by T cell proliferation were basically consistent with that of IFN-γ ELISPOT. In addition, T cell proliferation induced by No.53 and No.64 peptides was found in the donor 1 and 4 respectively, but not IFN-γ secreting. Conclusion No.51 and No.70 peptide could be T cell epitope on C-terminal of nucleocapsid protein.
出处
《中华微生物学和免疫学杂志》
CAS
CSCD
北大核心
2004年第5期393-396,共4页
Chinese Journal of Microbiology and Immunology
