【摘要】 目的通过前S1合成肽对肝细胞癌(HCC)组织中浸润T淋巴细胞亚群表达的分析,探讨前S1合成肽对乙型肝炎病毒(HBV)导致的HCC组织中浸润T淋巴细胞亚群表达和CD4+/CD8+比值的影响,为特异性治疗HCC提供依据。方法用Merrifield固相化学合成法合成HBV中前S1抗原性最强的P34～59多肽段。测序后,用0.01mol/L磷酸盐缓冲液溶解除菌后作为抗原。选择12例术后病理诊断为HCC,术前血清乙肝标志物检测HBsAg、HBeAg、抗-HBc阳性和HBVDNA阳性,且HBsAg在HCC组织中表达亦为阳性的病例作为研究对象。在96孔培养板的每孔加入经密梯度离心获得的单个核细胞悬液100μl(1×106细胞),设每孔含前S1合成肽的量分别为1、5、10、15μg及白细胞介素-II500u和阴性对照共6组,每组均设3复孔,作用7天后制片,采用APAAP法观察CD3+、CD4+、CD8+和CD4+/CD8+的比值变化。结果含前S1合成肽10μg/孔和15μg/孔组中,CD4+表达有增加,分别为(35.3±3.4)%和(34.9±3.8)%,与自身对照[(29.3±3.5)%]相比差异有统计学意义(t=4.839,P<0.01;t=5.389,P<0.01);CD4+/CD88+比值两组分别为1.09±0.39,1.14±0.43,与自身对照0.81±0.41相比差异有统计学意义(t=4.041,P<0.01;t=4.762,P<0.01);10μg/孔和15μg/孔组CD4+表达和CD4+/CD8+比值差异无统计学意义(t=0.289,P>0.05;t=0.361,P>0.05)。结论在适当的剂量下,前S1合成肽能够在体外改变HCC组织中T淋巴细胞亚群的表达,使CD4+增加,但不存在剂量效应关系,同时能改变CD8+静息状态,使二者互相作用,达到杀伤肿瘤细胞的目的 ,为特异性运用前S1蛋白提供客观依据。更多还原

【Abstract】 Objective To explore whether PreS1 synthetic peptides can change the expression of T lymphocyte subgroups and ratio of CD+44/CD+8 in hepatocellular carcinoma (HCC) caused by hepatitis B virus (HBV).Methods P34-59 region composed by way of Merrifield solid phase synthesis,which was the most intensive antigen in PreS1 peptides,served as the antigen after dissolved in 0.01mol/L PBS.Twelve patients were chosen as objects,which were pathologically diagnosed as HCC after operation,with HBsAg,HBeAg,anti-HBc and HBV DNA positive in serum as well as HBsAg positive in HCC tissue.Monocytes were isolated and cultured in 96-well cell culture plate with 1×106 cells in every well.Then PreS1 synthetic petides were added in dose of 1μg,5μg,10μg and 15μg,while IL-2 of 500 U was also added in.Seven days later,percentage of CD+3,CD+4,CD+8 and ratio of CD+4/CD+8 were detected.Results Percentage of CD+4 increased significantly (t = 4.839,P < 0.01;t = 5.389,P < 0.01).Ratio of CD+4/CD+8 decreasedly significantly (t = 4.041,P < 0.01;t = 4.762,P < 0.01) in two groups where PreS1 synthetic peptides were added in dose of 5μg and 10μg.The above-mentioned items were compared with those in control group.The percentage of CD+3 in IL-2 group rised significantly compared with that in control group.Conclusion With proper doses,PreS1 synthetic peptides can change the expression of T lymphocyte subgroups in HCC tissue,increase the percentage of CD+4 obviously and change the quiescent condition of CD+8 to make carcinoma cells be killed through the interaction of CD+4 and CD+8.更多还原