HBV变异患者外周血CD4^＋CD25^＋调节性T细胞变化及其临床意义

Relationship between CD4^+CD25^+ regulatory T cells and exacerbation of liver disease related HBV mutation patients

目的研究乙型肝炎病毒(HBV)变异患者外周血CD4+CD25+调节性T细胞(Treg细胞)及肝组织CD8+T细胞的关系,并探讨其临床意义。方法慢性乙型肝炎患者64例。HBV变异者27例,无变异者37例;急性乙型肝炎患者16例。各组间年龄、性别无明显差异。25例患者进行肝脏穿刺活检,免疫组化方法检测肝组织CD8+T细胞分布;流式细胞技术测定外周血Treg细胞变化;荧光定量PCR测定血清HBVDNA水平,PCR-RFLP测定前C区G1896A、P区YMDD变异。结果急性乙型肝炎患者、HBV变异伴肝病进展患者外周血Treg细胞比例明显低于HBV变异肝病无明显变化者(3.00±1.33,2.57±0.83vs4.32±0.96,P<0.01),后者与HBV无变异者没有明显差异(4.32±0.96vs4.77±2.11,P>0.05)。外周血Treg细胞与HBVDNA水平呈正相关(r=0.411,P<0.01);免疫组化显示HBV变异伴肝病进展患者肝组织CD8+T明显增多,急性乙型肝炎患者肝组织汇管区较多CD8+T浸润。结论Treg比例降低与HBV变异肝病进展可能有关,可作为HBV变异后临床转归的预测指标。初步显示,肝组织CD8+T浸润与外周血Treg细胞呈负相关,提示HBV变异后肝脏病进展的免疫损伤机制。

Objective To study the relationship Between circulating CD4^ ＋ CD25^ ＋ Regulatory T （Tregs） Cells and hepatic expression of CD8 ^＋ T in HBV mutation patients. Its clinical significance was also surveried. Methods A total of 80 subjects, including 64 chronic hepatitis B patients （CHB）, 16 acute hepatitis B patients（AHB） were enrolled in the study. Of CHB patients, 27 patients with HBV mutation （22 with natural Pre C mutation, 5 with YMDD mutation related Lamivudin）. The gender and average age were comparable between CHB and AHB patients. The Tregs were determined by FCM. The hepatic expression of CD8^ ＋ T were measured by immunohistochemistry stain. The quantity HBV - DNA and HBV mutation were detected by PCR. Results The frequency of Tregs in AHB and HBV mutation with exacerbation of liver disease patients were decreased, compared to that in CHB without HBV mutation （3.00 ±1.33,2. 57 ±0. 83 vs 4. 77 ±2. 11 ,P 〈 0. 01 ）. However, the frequency of Tregs in HBV mutation without exacerbation of liver disease patient were comparable to that in CHB without HBV mutation patients （4. 32 ±0. 96 vs 4. 77 ± 2. 11, P 〉 0. 05 ）. The frequency of Tregs cells was positively correlated with serum HBV DNA load. The hepatic CD8 ^＋T expression were significantly incresesd in AHB and HBV mutation with exacerbation of liver disease patients. The negtive relationship between hepalic CD8^ ＋ T and circulating Treg was also observed. Conclusion The positive relationship between the decreased Tregs and exacerbation of liver disease in patients with HBV mutation is eertified. The inereased hepatic expression of Cd8^ ＋ T may be contributed to exacerbation of liver disease. This may play an important role in HBV mutation result in immune hepatic injury.