摘要
目的建立HLA-A*2402限制的HBcAg特异性CTL细胞克隆。方法取HLA-A*2402阳性的慢性HBV感染者的PBMC,用限制性表位肽(HBV core117-125,EYLVSFGVW)和重组HBcAg刺激,并以有限稀释法对活化的T细胞进行克隆化,用免疫荧光染色、流式细胞术以及乳酸脱氢酶释放实验对所获克隆进行鉴定。结果PBMC经表位肽激活后,特异性细胞毒活性达到43.7%;从活化细胞获得11株T细胞克隆;除2株是CD4+T细胞克隆外,其余9株为CD8+T细胞克隆,其中2株为Tc1,3株为Tc2和4株为Tc0。9株CD8+T克隆在效/靶比例为2.5:1时均具有HBcAg特异性靶细胞溶解活性(33.9%～90.2%)。结论用HBVcore117-125能激活HLA-A*2402阳性的慢性HBV感染者外周血CD8+T细胞,随之建立的9株CD8+T克隆均具有HBcAg表位特异性细胞毒活性。
Objective To establish HLA-A＊2402 restricted HBV core antigen（HBcAg）-specific CTL clones.Methods PBMC obtained from a patient with chronic hepatitis B with HLA-A＊2402 molecule positive were stimulated by using synthetic peptides（HBc117-125,EYLVSFGVW） and rHBcAg,and T cell clones subsequently were established by limiting dilution technique.T cell clones were then characterized by immunofluorescence staining,flow cytometry analysis and LDH release.Results After 3wk of in vitro stimulation,PBMC from the patient displayed peptide-specific cytolytic activity（43.7%,E/T ratio=60：1）;Eleven HBV-specific T cell clones were established from these activating PBMC by limiting dilution technique;Nine of these clones were CD8＋T clones,and the other two were CD4＋T clones;The nine CD8＋T cell clones,including 2 Tc1,3 Tc2 and 4 Tc0,displayed peptidespecific cytolytic activity（33.9% to 90.2%,E/T ratio=2.5：1）.Conclusion HBc117-125 is capable of inducing Agspecific cytotoxic T cells in PBMC of HLA-A＊2402 positive patient with chronic hepatitis B.The Nine CD8 ＋T cell clones display HBcAg epitope-specific cytolytic activity.
出处
《实用肝脏病杂志》
CAS
2010年第2期85-88,共4页
Journal of Practical Hepatology
基金
国家自然科学基金资助项目(编号:30671929)
