原发性胆汁性肝硬化HLA-A* 0201限制性PDC-E2特异性CTL功能分析

Functional analysis of HLA-A* 0201-restricted PDC-E2 specific CTLs in primary biliary cirrhosis

目的:探讨原发性胆汁性肝硬化(PBC)中HLA-A* 0201限制的PDC-E2159-167特异性CTL与PDC-E2165-174特异性CTL功能。方法:分析15例HLA-A*0201阳性(A2+)PBC患者抗原肽PDC-E2159-167、PDC-E2165-174诱导的CTL对以表达HLA-A* 0201分子的T2细胞作为靶细胞的细胞毒性作用,并利用四聚体技术结合细胞内因子染色研究PDC-E2159-167特异性CTL与PDC-E2165-174特异性CTL分泌细胞因子(IFN-γ、IL-4和TNF-α)情况。结果:15例A2+PBC患者中,12例PDC-E2159-167诱导的CTL和10例PDC-E2165-174诱导的CTL对负载相应抗原肽的靶细胞特异杀伤活性大于10%(E∶T=40∶1),而对照组则均小于10%;15例A2+PBC患者从PBMC诱导的PDC-E2159-167特异性CTL和PDC-E2165-174特异性CTL中分泌IFN-γ细胞比值分别为0.33%±0.24%(0%～0.86%)、0.27%±0.24%(0%～0.75%),未检测到分泌TNF-α或IL-4的PDC-E2159-167或PDC-E2165-174特异性CTL。结论:结果表明,大部分PBC患者中HLA-A* 0201限制性PDC-E2159-167特异性CTL和PDC-E2165-174特异性CTL具有特异杀伤活性,HLA-A* 0201限制性PDC-E2特异性CTL主要分泌IFN-γ并非IL-4、TNF-α。

Objectlve：To investigate the effector functions of either HLA-A ＊ 0201-restricted PDC-E2159-167 specific CTLs or PDC-E2165-174 specific CTL in primary biliary cirrhosis, nethods：Cytotoxicity of PDC-E2159-167 or PDC-E2165-174-induced CTL lines from 15 cases of HLA-A＊ 0201 positive（ A2^＋） PBC patients was analyzed against T2 cells as compared to 3 A2^＋ patients with other clinical liver diseases and 3 A2^＋ healthy volunteers. Cytokines including IFN-γ, IL-4 and TNF-α produced by the PDC-E2 specific CTLs were detected with tetramer technology and intraceUular staining. Results： Of the 15 A2^＊ PBC patients, PDC-E2159-167 induced CTLs from 12 patients and PDC-E2165-174 induced CTLs from 10 patients, which showed specific cytotoxicity more than 10%, but all the controls were less than 10%. Frequencies of IFN-γ producing PDC-E2159-167 specific CTLs and PDC-E216-174 specific CTLs in A2^＋ PBC patients were 0. 33% ±0. 24% （0-0.86% ） and 0.27% ±0. 24% （0-0.75% ）, respectively, whereas production of IL-4 or TNF-α could not be detected after stimulated with PDC-E2 peptides. Conclusion：Our data suggest that HLA-A^＊ 0201 restricted PDC-E2159-167 or PDC-E2165-174 induced CTLs in most of A2^＋ PBC patients show specific cytotoxicity and produce cytokine IFN-γ, but not IL-4 and TNF-α.