Department of Immunology, Erasmus MC, University Medical Centre, Rotterdam, Netherlands.
Division of Rheumatology, Department of Paediatrics, The Hospital for Sick Children, Toronto, ON, Canada.
Front Immunol. 2019 Aug 29;10:2084. doi: 10.3389/fimmu.2019.02084. eCollection 2019.
Quantification of T-cell receptor excision circles (TRECs) has impacted on human T-cell research, but interpretations on T-cell replication have been limited due to the lack of a genomic coding joint. We here overcome this limitation with multiplex TRG rearrangement quantification (detecting ~0.98 alleles per TCRαβ+ T cell) and the HSB-2 cell line with a retrovirally introduced TREC construct. We uncovered <5 cell divisions in naive and >10 cell divisions in effector memory T-cell subsets. Furthermore, we show that TREC dilution with age in healthy adults results mainly from increased T cell replication history. This proliferation was significantly increased in patients with predominantly antibody deficiency. Finally, Guthrie cards of neonates with Down syndrome have fewer T and B cells than controls, with similar T-cell and slightly higher B-cell replication. Thus, combined analysis of TRG coding joints and TREC signal joints can be utilized to quantify T-cell replication, and has direct applications for research into aging, immunodeficiency, and newborn screening.
T 细胞受体切除环 (TREC) 的定量分析对人类 T 细胞研究产生了影响，但由于缺乏基因组编码接头，对 T 细胞复制的解释一直受到限制。我们通过使用多重 TRG 重排定量（检测每个 TCRαβ+T 细胞中的~0.98 个等位基因）和带有逆转录病毒引入的 TRECs 构建体的 HSB-2 细胞系克服了这一限制。我们发现，在幼稚 T 细胞亚群中，<5 个细胞分裂，在效应记忆 T 细胞亚群中，>10 个细胞分裂。此外，我们表明，健康成年人中 TREC 随年龄的稀释主要归因于 T 细胞复制史的增加。在主要抗体缺陷的患者中，这种增殖显著增加。最后，唐氏综合征新生儿的 Guthrie 卡的 T 细胞和 B 细胞比对照组少，T 细胞复制相似，B 细胞复制略高。因此，TRG 编码接头和 TRECs 信号接头的联合分析可用于定量 T 细胞复制，并直接应用于衰老、免疫缺陷和新生儿筛查的研究。
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