Measuring T cell receptor and T cell gene expression diversity in antigen-responsive human CD4+ T cells
Measuring T cell receptor and T cell gene expression diversity in antigen-responsive human CD4+ T cells
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DOI：
10.1016/j.jim.2013.11.003
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发表时间：
2013-12-31
影响因子：
2.2
通讯作者：
Bonifacio, Ezio
中科院分区：
文献类型：
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作者：
Eugster, Anne;Lindner, Annett;Bonifacio, Ezio
T细胞在TCR、表位识别和细胞因子产生方面具有多样性，可用于免疫监测。此外，TCR家族在疾病中的克隆扩增可能为TCR定向治疗提供机会。我们开发了单细胞TCR α和β链表达基因测序方法，并将其应用于疫苗（破伤风-类毒素）应答CD4(+) T细胞。在产生677种不同受体的3038个CD4(+) T细胞中，1309个（43%）成功测序了TCR α链和β链。破伤风类毒素应答细胞和非破伤风类毒素应答细胞的trv和TRBV基因使用差异（p = 0.004和0.0002），个体内破伤风类毒素应答细胞的TCR存在广泛的多样性。从同一受试者的不同样本中可以回收相同的TCR：强化疫苗接种后鉴定的TCR在强化前记忆T细胞中很常见（占强化前TCR的31%），在强化前接种初始细胞中也有鉴定（6.5%）。受试者之间没有共享TCR，但在受试者体内和受试者之间观察到破伤风类毒素应答细胞共享其TCR链之一。将单细胞基因表达谱与TCR测序相结合，揭示了具有相同TCR的细胞中不同细胞因子谱的例子。新的分子方法证明了个体内部和个体之间ag反应性CD4(+) T细胞的广泛多样性。(C) 2013 Elsevier B.V.版权所有
T cells have diversity in TCR, epitope recognition, and cytokine production, and can be used for immune monitoring. Furthermore, clonal expansion of TCR families in disease may provide opportunities for TCR-directed therapies. We developed methodology for sequencing expressed genes of TCR alpha and beta chains from single cells and applied this to vaccine (tetanus-toxoid)-responsive CD4(+) T cells. TCR alpha and beta chains were both successfully sequenced in 1309 (43%) of 3038 CD4(+) T cells yielding 677 different receptors. TRAV and TRBV gene usage differed between tetanus-toxoid-responsive and non-responsive cells (p = 0.004 and 0.0002), and there was extensive TCR diversity in tetanus-toxoid-responsive cells within individuals. Identical TCRs could be recovered in different samples from the same subject: TCRs identified after booster vaccination were frequent in pre-booster memory T cells (31% of pre-booster TCR), and also identified in pre-booster vaccination naive cells (6.5%). No TCR was shared between subjects, but tetanus toxoid-responsive cells sharing one of their TCR chains were observed within and between subjects. Coupling single-cell gene expression profiling to TCR sequencing revealed examples of distinct cytokine profiles in cells bearing identical TCR. Novel molecular methodology demonstrates extensive diversity of Ag-responsive CD4(+) T cells within and between individuals. (C) 2013 Elsevier B.V. All rights reserved.