HBV相关慢加急性肝衰竭患者Treg细胞、Th17细胞表达特点及其与HBV前C区/BCP区变异的相关性

摘要

Objective To investigate the expression characteristics of peripheral T regulatory cell (Treg) and T help cell 17 (Th17) in patients with hepatitis B related acute-on-chronic liver failure (HBV-ACLF),and their correlation with HBV pre-C/BCP mutations.Methods Forty-one HBV ACLF patients and 41 early ACLF patients were enrolled.Levels of serum interleukin (IL)-17,transforming growth factor (TGF)-β1,interferon (IFN)-γ,IL-1β and IL 21 were determined using enzyme-linked immuno sorbent assay (ELISA).Expression of Treg and Th17 were determined using flow cytometry.Sequencing for mutations of HBV pre-C/BCP region was performed.Results Levels of Treg,Th17,IL-17,TGF-β1,IFN-γ and IL-1β,A1762T/G1764A double mutations and G1896A mutation were significantly higher in HBV-ACLF patients than those in early HBV ACLF patients (t'=8.594,P＜0.001;t =4.609,P＜0.001;t =2.316,P-0.023;t =2.939,P =0.004;t =8.094,P＜0.001;t =5.429,P＜0.001;x2 =4.038,P =0.044;x2 =6.032,P =0.01),respectively.There were no significant difference in the ratio of Treg/Th17 and IL-21 level between HBV-ACLF and early HBV-ACLF group (t =0.902,P =0.370;t =0.294,P =0.769).Treg and Th17 levels in patients with A1762T/G1764A double mutations or G1896A mutation were both significantly higher than those with no mutations (t =2.932,P =0.004;t =2.339,P=0.022;t =3.232,P =0.002;r=2.990,P =0.004).][here were no significant difference in the ratio of Treg/ Th17 between mutation and non mutation groups (t =0.030,P =0.976;t'=0.272,P =0.787).Conclusion Changes of Treg and Th17,A1762T/G1764A double mutations in HBV BCP region and G1896A mutation in pre-C region might play roles in pathogenesis of HBV ACLF.%目的 探讨HBV相关慢加急性肝衰竭患者外周血调节性T淋巴细胞(Treg)、辅助性T淋巴细胞(Th)17细胞表达特点及其与HBV前C区/BCP区变异的相关性.方法 选择HBV相关慢加急性肝衰竭和肝衰竭前期患者各41例,采用ELISA检测外周血IL-17、TGFβ1、IFN γ、IL-1β及IL-21水平,流式细胞术检测Treg和Th17表达水平,直接测序法检测HBV前C区及BCP区基因,统计学处理用t检验.结果 肝衰竭患者Treg、Th17、IL-17、TGF-β1、IFN-γ、IL-1β水平、HBV变异位点中A1762T/G1764A双突变、G1896A突变比例分别为(9.28±1.12)％、(0.23±0.05)％、(151.83±35.22) pg/mL、(153.24±34.15) pg/mL、(177.61±29.25) pg/mL、(77.46±13.15) pg/mL、22/19、23/18,均高于肝衰竭前期患者的(7.47±0.74)％、(0.19±0.03)％、(134.02±34.39) pg/mL、(130.76±35.13) pg/mL、(130.68±22.87) pg/mL、(60.12±15.67) pg/mL、13/28、12/29,差异均有统计学意义(t’值分别为8.594、4.609、2.316、2.939、8.094、5.429,x2值分别为4.038、6.032,均P＜0.05);而Treg/Th17比值、IL-21水平在两组中差异无统计学意义(t=0.902,P=0.370;t=0.294,P=0.769).HBV BCP区A1762T/G1764A双突变和G1896A突变的患者Treg、Th17水平均高于野生型,差异有统计学意义(t=2.932,P=0.004;t=2.339,P=0.022;F 3.232,P=0.002;t'=2.990,P=0.004),而Treg/Th17比值在突变型和野生型中差异均无统计学意义(t=0.030,P=0.976;t’=0.272,P=0.787).结论 Treg细胞及Th17细胞变化,HBV BCP区A1762T/G1764A双突变、G1896A突变可能与HBV相关慢加急性肝衰竭相关.