汉滩病毒核蛋白诱导肾综合征出血热患者的T淋巴细胞反应

T cells responses induced by nucleocapsid protein of Hantaan Virus in patients of Hemorrhagic Fever with Renal Syndrome

目的测定汉滩病毒(HTNV)核蛋白(NP)C-端多肽诱导肾综合征出血热(HFRS)患者的特异性T淋巴细胞反应,为HFRS发病机制、疫苗研制及抗病毒免疫反应研究提供资料。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-γ酶联免疫斑点试验(ELIspot)测试13名患者对23条多肽的T淋巴细胞反应,筛选出反应较强的肽段。结果5名患者有不同程度的肽特异性T细胞反应。No.70肽可诱导3号和10号患者分泌IFN-γ,T细胞频率分别为51和55SFC/106PBMC,No.51肽可诱导4、7、12号患者分泌IFN-γ,T细胞频率分别为90、65、95SFC/106PBMC。结论No.51肽和No.70肽可诱导较强的T淋巴细胞反应,可能是NPC端的细胞毒性T淋巴细胞(CTL)表位。

[Objective] To detect T cell resposes to C-terminal of Hantaan Virus （HTNV） nucleocapsid protein （NP） and to lay the foundation for an understanding of Hemorrhagic Fever with Renal Syndrome（HFRS） pathogenesis, the design of vaccine strategies and anti-viral immune response. [Methods] Peripheral blood mononuclear cells （PBMCs） were separated on Ficoll-Hypaque density gradient centrifugation from HFRS patients. Using IFN-γ enzyme-linked immunospot （ELIspot） assay, T cell responses to 23 polypeptides on NP were examined from 13 patients with HFRS in the convalescent phase. The polypeptides with a stronger response were sorted out. [ Results ] IFN-γ ELIspot analysis showed that, of the 13 individuals tested, five had detectable HTNV-speeific T celt responses to NP peptides. No.70 peptide can induce secretion of IFN-γ in No.3 and No.10 patient. The frequency of peptide-speeific T cell was 51 and 55 SFC/10^6 PBMCs, respectively. No.51 peptide can induce secretion of IFN-γ in No.4,No.10 and No.12 patient. The frequency of peptide-specifie T cell/was 90, 65 and 95 SFC/10^6 PBMCs, respectively. [ Conclusion] No.51 and No.70 peptide can induce stronger T cell responses. They may be cytotoxie T lymphocyte （CTL） epitope in C-terminal of nucleoeapsid protein.