【摘要】 目的　研究慢性乙型肝炎患者外周血中可作用于病毒感染靶细胞的特异性CD3+ T细胞比率。方法　筛选HLA A2 患者 :将MHC :Ig双体融合蛋白试剂 ,加入HBV核心蛋白 (C)、聚合酶蛋白 (P)及表面抗原蛋白 (S)中HLA A2 限制性合成肽 ,与患者CD3+ 细胞作用 ,采用双重荧光抗体染色 ,用FACS检测各患者的特异性T细胞比率 ,同时设立各种对照管。结果　 2 3/ 5 4例 (42 .6 % )患者为HLA A2 。其中 15例经用HBVC、P、S肽染色后 ,5例患者的特异性CD3+ 阳性细胞比率高于对照管 ,4例患者有结合S肽的特异性CD3+ 细胞 ,其中 1例同时可结合P肽 ,1例仅与P肽结合。结论　慢性乙型肝炎患者外周血中与特异性肽结合的CD3+ T细胞比率较少。MHC :Ig双体融合蛋白技术使用方便 ,但非特异性染色率偏高。本技术可用于随访治疗中的慢性乙型肝炎患者免疫状态的变化更多还原

【Abstract】 Objective To study the percentage of HBV specific CD3 + T cells in the peripheral blood of chronic hepatitis B patients. Methods HLA A 2 patients were screened by MHC:Ig dimer reagent. HLA A 2 restricted HBV core, polymerase, and surface peptides were added to T cells of chronic hepatitis B patients, followed by double staining with anti CD3 antibodies and antibodies reactive with the heavy chain of Ig. FACS was used for monitoring HBV specific T cell percentage. Results 23/54 (42.6%) patients belonged to HLA A 2. Fifteen of them were reacted with HBV peptides. CD3 + T cells of five patients showed higher percentage of binding with the HBV peptides than that in the controls. Four patients showed CD3 + T cells binding with S peptide, one with P peptide and one binding with both S and P peptide. Conclusions The percentage of T cells which could bind with HBV peptide was low in chronic hepatitis B and the binding efficiency was low as well. The MHC:Ig dimer reagent is convenient for counting the number of HBV specific T cells, but the nonspecific staining background seems to be relatively high. However, this technology is useful to monitor changes of immune responses in chronic hepatitis B patients during immuno therapy.更多还原