Department of Neonatology, Longgang District Central Hospital of Shenzhen, Shenzhen, Guangdong 518116, China.
Zhongguo Dang Dai Er Ke Za Zhi. 2021 Nov 15;23(11):1154-1160. doi: 10.7499/j.issn.1008-8830.2106044.
To investigate the diversity of peripheral blood T cell receptor (TCR) β chain complementarity-determining region 3 (CDR3) based on immune repertoire sequencing in neonates with sepsis and the possible pathogenesis of neonatal sepsis.
A total of 12 neonates with sepsis were enrolled as the case group, and 9 healthy full-term infants, matched for gestational age, birth weight, and age, were enrolled as the control group. Omega nucleic acid purification kit (SQ blood DNA Kit II) was used to extract DNA from peripheral blood samples, TCR β chain CDR3 was amplified by multiplex PCR, and then high-throughput sequencing was performed for the products to analyze the diversity of TCR β chain CDR3 and the difference in expression.
The length and type of TCR β chain CDR3 were similar between the case and control groups, and Gaussian distribution was observed in both groups. With D50 and Shannon-Wiener index as the evaluation indices for diversity, the case group had a significantly lower diversity of TCR β chain CDR3 than the control group (<0.05). The frequency of 48 genes in TCR β chain V segment was compared, and the results showed that compared with the control group, the case group had significantly higher frequencies of , , and (<0.05). The frequency of 13 genes in TCR β chain J segment were compared, and the results showed that compared with the control group, the case group had significantly higher frequencies of , , and (<0.05).
There is a significant change in the diversity of TCR β chain CDR3 in the peripheral blood of neonates with sepsis, suggesting that it might be associated with the immune pathogenesis of neonatal sepsis.
通过免疫受体库测序技术研究脓毒症新生儿外周血 T 细胞受体(TCR)β链互补决定区 3(CDR3)的多样性,探讨其与新生儿脓毒症发病机制的关系。
选取 12 例脓毒症新生儿作为病例组,另选取同期出生、胎龄、出生体重及日龄相匹配的 9 例健康足月新生儿作为对照组。采用 Omega 核酸提取试剂盒(SQ 血 DNA Kit II)提取外周血 DNA,采用多重 PCR 扩增 TCRβ 链 CDR3,对产物进行高通量测序,分析 TCRβ 链 CDR3 的多样性及表达差异。
病例组和对照组 TCRβ 链 CDR3 的长度和类型相似,均呈正态分布。以 D50 和 Shannon-Wiener 指数作为多样性评价指标,病例组 TCRβ 链 CDR3 的多样性明显低于对照组(<0.05)。比较 TCRβ 链 V 区 48 个基因的频率,结果显示,与对照组相比,病例组中 、 、 的频率明显升高(<0.05)。比较 TCRβ 链 J 区 13 个基因的频率,结果显示,与对照组相比,病例组中 、 、 的频率明显升高(<0.05)。
脓毒症新生儿外周血 TCRβ 链 CDR3 多样性发生显著变化,可能与新生儿脓毒症的免疫发病机制有关。