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目的 探讨组织相容性抗原 (HLA) 肽四聚体流式细胞技术检测乙型肝炎病毒特异性CD8+ T细胞的临床应用价值。方法 采用HLA A2分子胞外段与乙型肝炎病毒 (HBV)核心表位肽core 18 2 7结合的HLA 肽四聚体 (Tetramer) ,设计流式细胞技术检测乙肝患者外周血中针对该肽段的特异性CD8+ T细胞数量 ,以占总计数CD8+ 细胞数的百分比表示。结果 11例HLA A2 + 的急性乙肝患者外周血特异性CD8+ T细胞为 0 0 2 %～ 2 0 4 % ,中位数为 0 2 0 % ,16例HLA A2 + 的慢乙肝患者为<0 0 1%～ 0 6 5 % ,中位数为 0 0 5 % ,二组之间差有显著性 (P <0 0 1)。 15例HLA A2 + 正常对照组、13例HLA A2 -急性乙肝对照组和 19例HLA A2 -慢性乙肝对照组的特异性的CD8+ T细胞均不超过0 0 2 %。结论 HLA 肽四聚体流式细胞技术能在体外直接检测HBV特异性的CD8+ T细胞数量 ,其水平一定程度上反映了特异性CTL应答状况 ,且与乙肝的不同临床转归有关。
Objective To investigate the value of histocompatibility antigen (HLA) peptide tetramer flow cytometry in detection of hepatitis B virus-specific CD8 + T cells. Methods Tetramer, HLA class A2 extracellular domain and core epitope binding peptide of hepatitis B virus (HBV) core 18 2 7, was designed and the flow cytometry Of specific CD8 + T cell numbers, expressed as a percentage of total counted CD8 + cells. Results The percentages of peripheral blood-specific CD8 + T cells in 11 patients with HLA A2 + were 0 0 2% -2 20% with a median of 0 2 0%, and 16 patients with chronic hepatitis B with HLA A2 + were <0 0 1% ~ 0 6 5%, the median was 0 0 5%, the difference between the two groups was significant (P <0.01). The specific CD8 + T cells in 15 cases of HLA A2 + normal control group, 13 cases of HLA A2 - acute hepatitis B control group and 19 cases of HLA A2 - chronic hepatitis B control group did not exceed 0.2%. Conclusion HLA peptide tetramer flow cytometry can directly detect the number of HBV-specific CD8 + T cells in vitro, and its level reflects the specific CTL response to some extent, which is related to the different clinical outcomes of hepatitis B patients.