摘要
目的 采用高通量测序的方法探讨脓毒症患者早期T细胞抗原受体（TCR）β链互补决定区3（CDR3）的组成及多态性。 方法 从深圳市人民医院重症医学科收集脓毒症患者6例，从体检科收集年龄匹配的对照患者9例，入科后留取其静脉血，分离外周血单核细胞并提取DNA，对TCR β链CDR3进行多重PCR扩增，然后对产物进行高通量测序及数据分析。 结果 脓毒症组（S组）和对照组（N组）的TCR β链CDR3长度呈高斯分布，且两组前60种特有核苷酸序列和特有氨基酸表达水平存在很大差异；针对TRβV片段59个基因的表达水平进行比较，其中TRBV 10-2、TRBV 10-3、TRBV 12-1、TRBV 5-7的表达水平差异有统计学意义（P＜0.05）；对TRβJ片段中的14个基因的表达水平进行了F检验，其中TRBJ 1-1、TRBJ 1-2、TRBJ 2-1、TRBJ 2-5的表达水平差异有统计学意义（P＜0.05）。 结论 脓毒症患者早期T细胞TCR β链CDR3的多态性发生了明显改变，提示其与脓毒症的免疫发病机制有关，这将为脓毒症的发病机制研究、治疗和预后的判断提供新的方法和手段。
Objective To analyze the composition and polymorphism of the T cell receptor （TCR） β-chain complement determining region（ CDR3 ） repertoire in the early stage of sepsis using high - throughput sequencing. Methods 6 cases of sepsis were included in the Intensive Care Unit in Shenzhen People＇s Hospital. 9 cases of age - matched volunteers were collected from the Physical Examination Center as control group. The venous blood was collected, the peripheral blood mononuclear cells were isolated and DNA was extracted. The TCR β - chain CDR3 was amplified by multiplex - PCR, and high -throughput sequencing and data analysis of the products were carried out. Results The length of TCR β- chain CDR3 of patients with sepsis and the control group exhibited a Gauss distribution, and there was a significant difference in the expression level of the first 60 specific nucleotide sequences and the specific amino acids between two groups. The expression level of the 59 genes of the TRβV fragment was compared, in which the expression level of TRBV 10 -2, TRBV 10 - 3, TRBV 12 - 1 and TRBV 5 - 7 were statistically different （ P 〈 0.05 ）. The expression level of 14 genes in TRβJ fragment was analyzed by F test. The expression level of TRBJ 1 - 1, TRBJ 1 - 2, TRBJ 2 - 1 and TRBJ 2 - 5 had significant differences （ P 〈 0.05 ）. Conclusion The polymorphism of TCR β - chain CDR3 in T cells in the early stage of sepsis has been significantly altered, suggesting that it isrelated to the immune pathogenesis of sepsis, which will provide a new method and means for the pathogenesis, treatment and prognosis of sepsis.
作者
江意春
袁丰华
赵颖
黄泽伟
刘振密
刘晓君
刘雪燕
Jiang Yi-chun;Yuan Feng-hua;Zhao Ying;Huang Ze-wei;Liu Zhen-mi;Liu Xiao-jun;Liu Xue-yan(Department of ICU,the Second Clinical Medical College（Shenzhen People＇s Hospital）,Jinan University,Shenzhen 518020,China)
出处
《中国急救医学》
CAS
CSCD
北大核心
2018年第11期976-981,I0001,共7页
Chinese Journal of Critical Care Medicine
