中国免疫学杂志2017，Vol.33Issue(6)：889-894,899,7.DOI:10.3969/j.issn.1000-484X.2017.06.017
乙型肝炎患者治疗前后外周血CD4+ CD25+ Tregs TCR CDR3谱系漂移特征分析
Spectrum drift characteristics of CD4+ CD25+ Tregs TCR CDR3 in hepatitis B patients before and after treatment
摘要
Abstract
Objective:To determine the spectrum drift characteristics of CI4+CD25+Tregs TCR β chain CDR3 in patients with different phases of acute hepatitis B (AHB) and chronic hepatitis B (CHB) patients before and after the entecavir treatment.Methods:Anticoagulation venous blood was collected from 4 normal control subjects,3 AHB patients with acute phase and convalescent phase,and 4 CHB patients before and after the entecavir treatment;and peripheral blood mononuclear cells were isolated;CD4+ CD25+ Tregs were separated by using the magnetic beads,and total RNAs were extracted from CD4+ CD25+ Tregs and used for reverse transcription.The TRBV CDR3 was amplified by polymerase chain reaction (PCR) with forward primers specific for 24 TRBV families and one fluorescence-labeled common reverse primer specific for the BC region.The PCR products were sent out for Genescan,and results were analyzed for the TRBV family CDR3 spectrum characteristics by using the Peak Scanner Software vl.0.Data were analyzed with the comparative t-test to perform the statistical analysis.Results:The CDR3 spectral types of the TRBV family showed drift characteristics in 3 cases of AHB patients with acute and convalescent phases;single/oligo peak spectral type family was observed in most of patients with acute phase;multiple peak spectral type was seen in patients with convalescent phase;and the common spectrum shift of TRBV4,10,14,16,19 families seen in patients with acute phase was changed to multiple peak spectral type.The clonal expansion of TRBV family in the CD4+CD25+Tregs in PBMC from AHB patients with convalescent phase was significantly lower than AHB patients with acute phase (t =9.456,P =0.011).The clonal expansion of Tregs TRBV13.2,15,16,18,20 family seen in C HB patients before treatment may interfere the virus removal through down-regulating the body's immune response;and with the decline of viral load in serum after the antiviral treatment,the clonal expansion of Tregs TRBV1,5.2,6,12,14,24 family may help body induce immune tolerance and result in the HBV persistence.The clonal expansion of TRBV family in the CI4+CD25+Tregs in PBMC from of CHB patients after antiviral treatment was increased (t =-0.666,P =0.553).Conclusion:TRBV4,10,14,16,19 family of spectrum shift seen in AHB patients with acute phase was changed to multiple peak spectral type in patients with convalescent phase,suggesting this transition may be associated with HBsAg and HBeAg turning to negative.The clonal expansion of Tregs TRBV13.2,15,16,18,20 family seen in CHB patients before treatment may interfere the virus removal through down-regulating the body's immune response;and with the decline of viral load in serum after the antiviral treatment,the clonal expansion of Tregs TRBV1,5.2,6,12,14,24 family may help body induce immune tolerance and result in the HBV persistence.关键词
慢性乙型肝炎/急性乙型肝炎/T淋巴细胞受体/互补决定区3/调节性T细胞/恩替卡韦Key words
Chronic hepatitis B/Acute hepatitis B/T-cell receptor/Complementarity determining region 3/CD4+ CD25+ Tregs/Entecavir分类
医药卫生引用本文复制引用
潘宗琴,吕红,庄勤建,姚新生,邱隆敏..乙型肝炎患者治疗前后外周血CD4+ CD25+ Tregs TCR CDR3谱系漂移特征分析[J].中国免疫学杂志,2017,33(6):889-894,899,7.基金项目
本文为贵州省社会发展公关基金资助项目(No.黔科合SY[2010]3049号). （No.黔科合SY[2010]3049号）
