乙肝相关性慢加急性肝衰竭合并细菌感染患者Treg/Th17细胞免疫表达的研究

Study on immune expression of Treg/Th17 cells in patients with HBV-ACLF and bacterial infection

目的观察乙型病毒性肝炎(简称乙肝)相关性慢加急性肝衰竭(hepatitis B virus related acute-on-chronic liver failure,HBV-ACLF)合并细菌感染患者外周血调节性T细胞(regulatory T cell,Treg)和辅助性T细胞17(helper T cell 17,Th17)细胞计数及相关细胞因子免疫表达。方法选择96例HBV-ACLF患者,依据感染情况分为有细菌感染组与无细菌感染组。采用流式细胞术检测患者外周血Treg、Th17细胞计数,细胞因子微球检测技术检测转化生长因子-β(transforming growth factor-β,TGF-β)、白介素-10(interleukin-10,IL-10)、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、白介素-17A(interleukin-17A,IL-17A)、白介素-23(interleukin-23,IL-23)表达,实时荧光定量PCR检测叉状头状转录蛋白3(forkhead box protein 3,Foxp3)、维甲酸相关孤核受体-γt(retinoid-related orphan nuclcar receptor-γt,ROR-γt)mRNA表达,及检测相关炎症指标[C反应蛋白(C-reactive protein,CRP)、降钙素原(procalcitonin,PCT)、内毒素(lipopolysaccharide,LPS)]水平。结果有细菌感染组Treg细胞计数与无细菌感染组比较,差异无统计学意义(P>0.05)。与无细菌感染组比较,有细菌感染组Th17细胞计数明显升高(P<0.01);有细菌感染组CRP、PCT、LPS水平均显著升高(P<0.01,P<0.05);有细菌感染组Th17、TNF-α、IL-17A、IL-23、ROR-γt mRNA表达均明显上调(P<0.01,P<0.05);有细菌感染组Foxp3 mRNA、IL-10表达均明显下调(P<0.01,P<0.05)。有细菌感染组恶化率(83.33%)显著高于无细菌感染组(11.11%)(P<0.01)。结论HBV-ACLF患者随着病程进展,感染风险增加,其感染后存在Th17细胞优势化现象,具体机制可能与Th17细胞过度增值分化导致炎症反应失控,加重肝组织炎症反应相关。

Objective To observe the cell count of regulatory T cell(Treg)and helper T cell 17(Th17)cell and the expression of related cytokines in peripheral blood of patients with hepatitis B virus related acute-on-chronic liver failure(HBV-ACLF)combined with bacterial infection.Methods 96 patients with HBV-ACLF were divided into bacterial infection group and non-bacterial infection group according to the infection.Flow cytometry was used to detect the cell count of Treg and Th17 in peripheral blood.The expression levels of transforming growth factor-β(TGF-β),interleukin-10(IL-10),tumor necrosis factor-α(TNF-α),interleukin-17A(IL-17A)and interleukin-23(IL-23)were detected by cytometric bead array.The expression of forkhead box protein 3(Foxp3)and retinoid-related orphan nuclcar receptor-γt(ROR-γt)mRNA were detected by real-time fluorescent quantitative PCR,and the levels of C-reactive protein(CRP),procalcitonin(PCT)and lipopolysaccharide(LPS)were detected.Results There was no significant difference in Treg cell count between the bacterial infection group and non-bacterial infection group(P>0.05).Compared with the non-bacterial infection group,Th17 cell count in bacterial infection group was significantly increased(P<0.01),CRP,PCT and LPS were significantly increased in the bacterial infection group(P<0.01,P<0.05).The expression of Th17,TNF-α,IL-17A,IL-23 and ROR-γt mRNA in the bacterial infection group were all up-regulated(P<0.01,P<0.05),Foxp3 mRNA and IL-10 expression were significantly down-regulated in bacterial infection group(P<0.01,P<0.05).The deterioration rate in the bacterial infection group(83.33%)was significantly higher than that in the non-bacterial infection group(11.11%)(P<0.01).Conclusion Patients with HBV-ACLF have an increased risk of infection with the progression of the disease,and there is a phenomenon of Th17 cell dominance after infection.It may be related to the excessive value-added differentiation of Th17 cell,which leads to the uncontrolled inflammatory response and aggravates the inflammatory response of liver tissue.