摘要
目的： 通过检测慢性乙型肝炎急性发作患者外周血Th1及Th2细胞的百分比及Th1/Th2的比值、乙型肝炎病毒（HBV）DNA载量以及肝功能的状况，了解在乙型肝炎病毒复制及肝细胞受损时Th1/Th2细胞免疫应答的变化情况 。
方法：研究对象为正常对照组19例，慢性乙型肝炎急性发作组17例。应用流式细胞仪测定外周血中淋巴细胞胞浆内含白细胞介素-4（IL-4）及干扰素-γ（IFN-γ）的细胞的百分比及比值，采用荧光定量聚合酶链反应技术（FQ-PCR），检测慢性乙型肝炎患者发作时及治疗后外周血清中HBV DNA的载量；应用全自动生化分析仪检测患者急性发作及治疗后血清中丙氨酸氨基转移酶（ALT）、天门冬氨酸氨基转移酶（AST）的活性和血清总胆红素（TB）的含量。
结果：（1）在慢性乙型肝炎急性发作时Th1细胞百分比（14.52±4.86%）明显高于治疗后（9.33±2.72%）（P<0.05），也比正常对照组高（9.92±4.54%）（P<0.05）；急性发作时Th2细胞百分比（1.64±0.59%）与治疗后（1.81±0.75%）相比较无显著性差异(P>0.05),与正常对照组（2.058±0.985%）相比较也无显著性差异；急性发作时Th1/Th2比值比正常对照组高（P<0.05）,也比治疗后Th1/Th2比值高(P<0.05), 治疗后Th1/Th2比值与正常对照组相比较，无显著性差异（P>.05）；（2）慢性乙型肝炎急性发作时HBV DNA含量（经log转换后）为7.87±1.96copies/ml，治疗后为5.45±1.95 copies/ml，两者相比较有显著性差异(P<0.05)；（3）急性发作时ALT为467.52±103.46U/L，治疗后为137.66±53.29 U/L，两者相比较有显著性差异（P<0.05）；（4）Th1细胞比例与ALT活性呈正相关关系（r=0.432, P<0.05）；，Th1/Th2比值与ALT活性呈正相关关系(r=0.493,P<0.05); Th1细胞比例与HBV DNA含量及
肝功能其他指标均无明显相关关系。
结论：（1）慢性乙型肝炎急性发作时Th1细胞比例与肝炎活动程度呈正相关；（2）慢性乙型肝炎急性发作与Th1/Th2的失衡密切相关。（3）Th1细胞比例测定可作为动态监测肝炎活动的有效指标之一。
To investigate the immune response of Th1/Th2 cells to HBV duplication and subsequent injury of liver cells through detection of Th1/Th2 cells’ ratio, HBV DNA load and liver function indexes in peripheral blood of chronic hepatitis B patients during acute exacerbation.
Methods: 19 cases of normal controls and 17 cases of chronic hepatitis B patients experiencing acute exacerbation were recruited into this study. Using flow cytometry assay, Th1/Th2 ratio was estimated by quantification of cytokins (IL-4, r-IFN) inside the plasma of single CD4+ T cells. HBV DNA load in peripheral serum of chronic HBV infectants during and after acute exacerbation period was analyzed by polymerase chain reaction (PCR) assay combined with fluorescent probe (FQ-PCR). Liver function indexes (ALT，AST，TB) during acute exacerbation and after treatment was estimated by Hitachi 7170A auto-chemistry analyzer. Results: (1) The percentage of Th1 cells（14.52±4.86%） in chronic HBV infectants during acute exacerbation was higher than that these patients after treatment（9.33±2.72%）（P<0.05） and that of normal controls（9.92±4.54%）（P<0.05）. Similarly, the ratio of Th1/Th2 during acute exacerbation was higher than that of normal controls （P<0.05） and that these patients after treatment (P<0.05). There was no statistic difference of Th2 cells percentage between cases and controls（P>0.05）. Also, there was no statistic difference of Th1/Th2 ratio between normal controls and cases after treatment (P>0.05). There was no statistic difference of Th2 cells percentage between patients before（1.64±0.59%） and after treatment（1.81±0.75%）, and between patients （1.64±0.59%）and controls（2.05±0.98%）. (2) HBV DNA load of chronic HBV infectants during acute exacerbation (7.87±
1.96copies/ml,log transformed) was higher than that these patients after treatment(5.45±1.95)(P<0.05). (3) ALT activity during acute exacerbation (467.52±103.46U/L) were significant higher than that these patients after treatment (137.66±53.29 U/L) (P<0.05). (4) The ratio of both Th1 (r=0.432, P<0.05) and Th1/Th2 (r=0.493, P<0.05) were positively associated with ALT activity. Nevertheless, The percentage of Th1 cells was not significantly associated with either HBV DNA load or other liver function indexes.
Conclusions: (1) The percentage of Th1 cells in chronic HBV infectants during acute exacerbation is positively associated with the severity of hepatitis. (2) The aberration of Th1/Th2 ratio is closely associated with acute exacerbation of chronic HBV infectants. (3) The percentage of Th1 cells is potential biomarker for dynamic monitoring of hepatitis.
引文
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