原发性胆汁性肝硬化患者外周血T细胞表型及细胞因子的变化及意义

Significance and change of serum cytokines and T lymphocyte subsets in peripheral blood in patients with primary biliary cirrhosis

目的观察原发性胆汁性肝硬化(PBC)患者不同时期的T淋巴细胞表型及细胞因子变化,探讨其在PBC诊治中的价值及意义。方法研究分为PBC组(32例),正常对照组(对照组,20例)。以流式细胞仪检测外周血CD4+、CD8+、CD4+/CD8+、CD4+CD25+、CD4+CD28-、CD8+CD95+T淋巴细胞;以双抗体夹心酶联免疫吸附测定(ELISA)法检测外周血细胞因子IL-2I、L-4I、L-10、TNF-α水平。结果PBC患者活动期CD4+、CD4+/CD8+、CD4+CD28-、CD8+CD95+T细胞与对照组比较明显升高(39.71±7.62)vs(27.81±3.21)(、2.31±0.23)vs(1.53±0.71)、(1.54±0.98)vs(4.15±1.89)、(9.46±4.48)vs(22.72±7.67),而CD4+CD25+与对照组比较明显降低(1.68±1.32)vs(3.99±1.21)(P<0.05)。治疗后CD4+CD25+T细胞比治疗前明显升高(P<0.05),CD4+、CD4+/CD8+的比值、CD4+CD28-和CD8+CD95+T细胞比治疗前均明显降低(P<0.05)。肝功严重损伤组外周血:CD4+CD25+T细胞明显低于肝功能轻度损伤组(1.52±0.74 vs 1.89±0.91,P<0.05),而CD4+CD28-和CD8+CD95+T细胞比例明显高于肝功能轻度损伤者(4.34±1.92 vs 2.98±1.73,23.19±7.86 vs 20.16±7.48,均P<0.05);且CD4+CD25+T细胞数量与肝功能损伤指标碱性磷酸酶(ALP)、谷丙转肽酶(GGT)呈负相关(R分别为-0.328,-0.337,均P<0.05),CD4+CD28-和CD8+CD95+T细胞数量与肝功能损伤指标ALP、GGT呈正相关(R分别为0.326,0.318,0.459,0.512,均P<0.05)。PBC活动期细胞因子IL-2、TNF-α与对照组比较明显升高(166.54±9.05)ng/L vs(25.03±4.14)ng/L(、137.13±13.49)ng/L vs(47.66±9.73)ng/L(P<0.05)。治疗后IL-10和IL-4明显升高,而IL-2和TNF-α明显降低(P<0.05)。肝功严重损伤者IL-2和TNF-α水平明显高于肝功能轻度损伤者(P<0.05);且IL-2和TNF-a水平与肝功能损伤指标ALP和GGT呈正相关(R分别为0.426、0.436、0.514、0.512,P<0.05)。结论PBC患者活动期外周血以CD4+、CD4+CD28-和CD8+CD95+T细胞升高、CD4+CD25+降低和辅助T细胞亚群1(Th1)细胞因子占优势,治疗后以CD4+CD25+升高和辅助T细胞亚群2(Th2)细胞因子占优势,提示细胞免疫和细胞因子的免疫调节在PBC发病机制中发挥重要作用。

Objective To detect serum cytokines and T lymphocyte subsets in peripheral blood in patients with primary biliary cirrhosis（PBC） and evaluate the significance for diagnosis of PBC. Methods The subjects included PBC group（32 cases） and normal control group（control group,20 cases）. The number of CD4^＋,CD8^＋,CD4^＋CD28^- ,CD4^＋CD25^＋and CD8^＋CD95T cells were detected by flow cytometry and cytokines（IL-2, IL-4, IL-10, TNF-α） in the PBC patient serum by ELISA. Results CD4^＋, CD4^＋CD8^＋, CD4^＋CD28^- , CD8^＋CD95^＋T cells in PBC patients active stage were significantly higher than control group, respectively （39.71±7.62） vs （27.81± 3.21）, （2.31 ± 0.23） vs （1.53±0.71）,（4.15±1.89） vs （1.54±0.98）, （22.72±7.67） vs （9.46±4.48）. But CD4^＋CD25^＋decreased obviously（ P 〈0.05）, （1.68 ± 1.32） vs （3.99 ± 1.21） CD4^＋CD25^＋T cell increased obviously after therapy（ P〈 0.05）. CD4^＋CD8^＋ CD4^＋ CD4^＋CD28^- and CD8^＋CD95^＋T cell decreased obviously compared to before therapy （P 〈 0.05）. The peripheral blood CD4^＋CD25^＋T cells in the patients with severe hepatic damnification was significantly lower compared to those of patients without severe hepatic （1.52 ±0.74 vs 1.89± 0.91, P〈0.05）. The percentage of CD4^＋CD28^- and CD8^＋CD95^＋T ceils in the patients with severe hepatic damnification was significantly higher compared to that of the patients without severe hepatic （4.34±1.92 vs 3.98±1.73,23.19±7.86 vs 21.56±7.48,all P 〈 0.05）. CD4^＋CD25^＋T cell number of peripheral blood of PBC patients and liver functional damnification index（ALP, GGT） showed negative correlation （ r=-0. 328, -0. 337, respectively, all P〈0.05）. The numbers of CD4^＋CD28^-,CD8＋D95^＋T cell and liver functional damnification index （ALP, GGT） showed positive correlation （r = 0. 326,0. 318,0. 459,0. 512, respectively, all P 〈 0. 05）. The cytokines IL-2, TNF-α were respectively in PBC active stage increased obviously compared with those of control group （25.03±4.14） ng/L vs （166.54±9.05） ng/L, （47.66± 9.73） ng/L vs （137.13 ± 13.49） ng/L（ P 〈0.05）. IL-10, IL-4 increased and IL-2, TNF-α decreased obviously after therapy （ P 〈0. 05）. The level of IL-2 and TNF-α in the patients with severe hepatic damnification were significantly higher compared to those of the patients without severe hepatic damnification（ P 〈0.05）. And the level of IL-2,TNF-α and liver functional damnification index （ALP, GGT） showed positive correlation （ r = 0. 426,0. 436, 0. 514,0. 512, P 〈0.05）. Conclusion The increase of CD4^＋, CD4^＋CD28^- and CD8^＋CD95^＋T cell, CD4^＋CD25^＋decrease and Thl cytokines are dominated in the peripheral blood of PBC patients＇ active stage;CD4^＋CD25^＋decrease and Th2 cytokines are dominated after therapy. And the cell immunity and immunity adjustment of cytokines display very important effect in PBC disease mechanism.