HBsAg、HBsAb双阳性HBV感染者TCR β链CDR3克隆型分析

Analysis of TCR β chain CDR3 clonetype in HBV-infected patients with coexistence of HBsAg and HBsAb

目的分析HBsAg和HBsAb双阳性乙型肝炎患者与其他HBV感染者间T细胞受体(TCR)组库β链互补决定区3(CDR3)克隆型差异性。方法以11例HBsAg和HBsAb双阳性乙型肝炎患者为病例组,10例自然痊愈(HBsAb^+)者为对照组1,10例HBsAg阳性但HBsAb阴性乙型肝炎患者为对照组2。用Illumina HiseqX10测序仪对全血DNA的CDR3序列进行高通量测序,建立CDR3免疫组库,并进行CDR3克隆型及多样性分析。结果病例组任意两样本CDR3克隆型重叠率为6.28%(0.25%,13.10%);对照组1为10.49%(6.20%,17.30%);对照组2为2.60%(0.13%,13.69%),病例组与对照组1相比差异有统计学意义(P=0.008),对照组1与对照组2组相比差异有统计学意义(P=0.001),病例组与对照组2相比差异无统计学意义。经过病例组与2个对照组分别比较得到:克隆型TRBV7-2/TRBD1/TRBJ2-1频率病例组高于对照组1(P=0.029),克隆型TRBV7-3/TRBD1/TRBJ2-7频率病例组低于对照组1(P=0.031)。病例组与对照组1相比,V基因型TRBV5-8频率差异有统计学意义(P=0.047);病例组与对照组2之间,有14种克隆型频率差异有统计学意义,V基因型TRBV28频率差异有统计学意义(P=0.028)。3组样本的TCRβ链CDR3多样性差异无统计学意义(P>0.05)。结论克隆型TRBV7-2/TRBD1/TRBJ2-1和TRBV7-3/TRBD1/TRBJ2-7,V基因型TRBV5-8可能与HBsAg和HBsAb双阳性相关,而TCRβ链CDR3多样性与HBsAg和HBsAb双阳性无明显关系。

Objective To compare T-cell receptor (TCR)β chain complementarity-determining region 3 (CDR3) in the patients with coexistence of HBsAg and HBsAb and other HBV infected patients. Methods The clonotype and diversity of CDR3 in blood of group cases (positive HBsAg and HBsAb)( n =11), control 1 (negative HBsAg and positive HBsAb)( n =10) and control 2 (positive HBsAg and negative HBsAb)( n =10) were analyzed by high-throughput TCR sequencing with Illumina HiseqX10. Results In the case group, the overlap rate of 6.28%(0.25%, 13.10%) was detected between any two samples, which was significantly lower than the overlap rate of 10.49%(6.20%,17.30%) seen in control 1 group ( P =0.008). In control 2 group, the overlap rate of 2.60%( 0.13%,13.69%) was significantly lower than control 1 group ( P =0.001). There was no difference between case group and control 2 group. After pairwise comparison between the three groups, the frequency of clonotype TRBV7-2/TRBD1/TRBJ2-1 in case group was higher than that of control 1 group ( P =0.029), the frequency of TRBV7-3/TRBD1/TRBJ2-7 in case group was lower than that of control 1 group ( P =0.031). The difference of TRBV5-8 was significant in comparing case group with control 1 group ( P =0.047). There were 14 clonotypes which had differences between case group and control 2 group in frequency. TRBV28was significant in comparing case group with control 2 group ( P =0.028). For diversity, there was no difference among the three groups. Conclusion Clonotype TRBV7-2/TRBD1/TRBJ2-1, TRBV7-3/TRBD1/TRBJ2-7 and TRBV5-8 were associated with coexistence of HBsAg and HBsAb, but the diversity was not associated with TCR β chain CDR3.