慢性乙型肝炎患者抗病毒治疗前后HBeAg特异性T细胞应答特征研究

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目的研究慢性乙型肝炎(CHB)患者抗病毒治疗前后T细胞对HBV特异性抗原蛋白(HBeAg)的免疫应答特征。方法分别收集22例慢性乙型肝炎患者抗病毒治疗前、治疗后3月、6月和12月的外周血,以HBeAg蛋白刺激外周血单个核细胞,采用酶联免疫斑点技术(ELISPOT)检测产生IFN-γ的HBeAg特异性T细胞的频率和强度。结果 1)CHB患者抗病毒治疗前和治疗后3月、6月、12月总的T细胞反应频率分别为31.8%(7/22)、50.0%(11/22)、77.3%(17/22)和95.5%(21/22),治疗后12月反应频率明显高于治疗前和治疗后3月(P<0.05),治疗后6月反应频率明显高于治疗前(P<0.05),核苷类似物治疗组与干扰素治疗组治疗后T细胞反应频率均无明显差异(P>0.05)。2)治疗后12月T细胞的反应强度明显高于治疗前、治疗后3月及治疗后6月(P<0.05)。核苷类似物治疗组与干扰素治疗组均分别得出上述相同的结果。3)将患者T细胞的反应强度与HBV DNA载量(取对数值表示)做Spearman秩相关分析,rs=-0.186<0,P=0.041,P<0.05,认为患者T细胞对HBeAg蛋白的反应强度与HBV DNA载量存在负相关关系。结论 HBeAg可以刺激CHB患者产生特异性T细胞应答,且随着抗病毒治疗时间的延长,患者HBeAg特异性T细胞的应答频率和应答强度均有所增强,且与血清HBV DNA水平呈负相关关系。 Objective To investigate the immune response of T cells to HBV specific antigen protein (HBeAg) in patients with chronic hepatitis B (CHB) before and after antiviral therapy. Methods Peripheral blood mononuclear cells of 22 patients with chronic hepatitis B before antiviral therapy, 3 months, 6 months and 12 months after treatment were collected and stimulated by HBeAg protein respectively. ELISPOT IFN-γ HBeAg-specific T cell frequency and intensity. Results 1) The frequency of T cell responses in CHB patients before and after 3 months, 6 months and 12 months of treatment were 31.8% (7/22), 50.0% (11/22) and 77.3% (17/22 ) And 95.5% (21/22) respectively. The response frequency at 12 months after treatment was significantly higher than that before treatment and 3 months after treatment (P <0.05). The reaction frequency at 6 months after treatment was significantly higher than that before treatment (P <0.05) There was no significant difference in the frequency of T cell responses between glycosylated and interferon treated groups (P> 0.05). 2) The response intensity of T cells in December after treatment was significantly higher than that before treatment, 3 months after treatment and 6 months after treatment (P <0.05). The same results were obtained for the nucleoside analogue treatment group and the interferon treatment group, respectively. 3) Spearman rank correlation analysis was performed between the response intensity of T cells and HBV DNA load (expressed as logarithm), rs = -0.186 <0, P = 0.041, P <0.05. The response of patients with T cells to HBeAg protein There is a negative correlation between intensity and HBV DNA load. Conclusions HBeAg can stimulate specific T cell responses in patients with CHB, and with the prolongation of antiviral therapy, the response frequency and response strength of HBeAg-specific T cells are increased, and negatively correlated with serum HBV DNA levels .
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