HBsAg、HBsAb双阳性HBV感染者TCR β链CDR3克隆型分析
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摘要
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. TRBV28 was 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.
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. TRBV28 was 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.
引文
[1]Luo Z,Li L,Ruan B.Impact of the implementation of a vaccination strategy on hepatitis B virus infections in China over a 20-year period[J].Int J Infect Dis,2012,16(2):e82-e88.
[2]Liu Y,Zhang L,Zhou J Y,et al.Clinical and virological characteristics of chronic hepatitis B patients with coexistence of HBsAg and anti-HBs[J].PLoS One,2016,11(1):e146980.
[3]Soroosh P,Shokri F,Azizi M,et al.Analysis of T-cell receptor beta chain variable gene segment usage in healthy adult responders and nonresponders to recombinant hepatitis B vaccine[J].Scand J Immunol,2003,57(5):423-431.
[4]Yang J,Chen J,He J,et al.Profiling the repertoire of T-cell receptor beta-chain variable genes in peripheral blood lymphocytes from subjects who have recovered from acute hepatitis B virus infection[J].Cell Mol Immunol,2014,11(4):332-342.
[5]Hou X,Guo Q,Wei W,et al.Screening of genes related to early and late flowering in tree peony based on bulked segregant RNA sequencing and verification by quantitative real-time PCR[J].Molecules,2018,23(3):pii:E689.
[6]周婷婷,冯健男.抗体信息数据库[J].中国生物化学与分子生物学报,2017,33(8):750-755.
[7]Linninge C,Roth B,Erlanson-Albertsson C,et al.Abundance of Enterobacteriaceae in the colon mucosa in diverticular disease[J].World J Gastrointest Pathophysiol,2018,9(1):18-27.
[8]傅晓春,刘灿,欧启水.慢性乙型肝炎患者HBsAg和抗HBs共存的发生机制及其临床意义[J].临床检验杂志,2016,34(8):566-570.
[9]傅晓春,陈静,叶爱珠,等.医院就诊人群HBsAg与抗HBs双阳性HBV感染者的流行病学与分子病毒学特征[J].临床检验杂志,2017,35(1):47-52.
[10]Park GC,Hwang S,Ahn CS,et al.Analysis of S gene mutation of the hepatitis B virus in adult liver transplant recipients showing resistance to hepatitis B immunoglobulin therapy[J].Transplant Proc,2013,45(8):3047-3051.
[11]Ding F,Yu HG,Li YX,et al.Sequence analysis of the HBV S protein in Chinese patients with coexisting HBsAg and anti-HBs antibodies[J].J Med Virol,2015,87(12):2067-2073.
[12]Chen Y,Qian F,Yuan Q,et al.Mutations in hepatitis B virus DNA from patients with coexisting HBsAg and anti-HBs[J].J Clin Virol,2011,52(3):198-203.
[13]付钰,杨国胜,吴松.免疫组库的临床应用[J].国际免疫学杂志,2014,37(5):411-414,423.
[14]王鹏,贺晓燕,王小妹,等.T细胞TCR CDR3受体库的高通量测序分析概况[J].现代免疫学,2011,31(6):512-516.
[15]Oliveira J,Mahony J,Hanemaaijer L,et al.Biodiversity of bacteriophages infecting Lactococcus lactis starter cultures[J].J Dairy Sci,2018,101(1):96-105.
[2]Liu Y,Zhang L,Zhou J Y,et al.Clinical and virological characteristics of chronic hepatitis B patients with coexistence of HBsAg and anti-HBs[J].PLoS One,2016,11(1):e146980.
[3]Soroosh P,Shokri F,Azizi M,et al.Analysis of T-cell receptor beta chain variable gene segment usage in healthy adult responders and nonresponders to recombinant hepatitis B vaccine[J].Scand J Immunol,2003,57(5):423-431.
[4]Yang J,Chen J,He J,et al.Profiling the repertoire of T-cell receptor beta-chain variable genes in peripheral blood lymphocytes from subjects who have recovered from acute hepatitis B virus infection[J].Cell Mol Immunol,2014,11(4):332-342.
[5]Hou X,Guo Q,Wei W,et al.Screening of genes related to early and late flowering in tree peony based on bulked segregant RNA sequencing and verification by quantitative real-time PCR[J].Molecules,2018,23(3):pii:E689.
[6]周婷婷,冯健男.抗体信息数据库[J].中国生物化学与分子生物学报,2017,33(8):750-755.
[7]Linninge C,Roth B,Erlanson-Albertsson C,et al.Abundance of Enterobacteriaceae in the colon mucosa in diverticular disease[J].World J Gastrointest Pathophysiol,2018,9(1):18-27.
[8]傅晓春,刘灿,欧启水.慢性乙型肝炎患者HBsAg和抗HBs共存的发生机制及其临床意义[J].临床检验杂志,2016,34(8):566-570.
[9]傅晓春,陈静,叶爱珠,等.医院就诊人群HBsAg与抗HBs双阳性HBV感染者的流行病学与分子病毒学特征[J].临床检验杂志,2017,35(1):47-52.
[10]Park GC,Hwang S,Ahn CS,et al.Analysis of S gene mutation of the hepatitis B virus in adult liver transplant recipients showing resistance to hepatitis B immunoglobulin therapy[J].Transplant Proc,2013,45(8):3047-3051.
[11]Ding F,Yu HG,Li YX,et al.Sequence analysis of the HBV S protein in Chinese patients with coexisting HBsAg and anti-HBs antibodies[J].J Med Virol,2015,87(12):2067-2073.
[12]Chen Y,Qian F,Yuan Q,et al.Mutations in hepatitis B virus DNA from patients with coexisting HBsAg and anti-HBs[J].J Clin Virol,2011,52(3):198-203.
[13]付钰,杨国胜,吴松.免疫组库的临床应用[J].国际免疫学杂志,2014,37(5):411-414,423.
[14]王鹏,贺晓燕,王小妹,等.T细胞TCR CDR3受体库的高通量测序分析概况[J].现代免疫学,2011,31(6):512-516.
[15]Oliveira J,Mahony J,Hanemaaijer L,et al.Biodiversity of bacteriophages infecting Lactococcus lactis starter cultures[J].J Dairy Sci,2018,101(1):96-105.