期刊
JOURNAL OF IMMUNOLOGICAL METHODS
卷 400, 期 -, 页码 13-22出版社
ELSEVIER SCIENCE BV
DOI: 10.1016/j.jim.2013.11.003
关键词
Human; T cells; T cell receptor; Gene expression
资金
- Deutsche Forschungsgemeinschaft-Center for Regenerative Therapies Dresden, Cluster of Excellence [FZ 111]
- Deutsche Forschungsgemeinschaft [DFG ZI-310/14-1, DFG ZI-310/14-2, DFG ZI-310/14-3, DFG ZI-310/14-4]
- German Ministry for Education and Research (BMBF)
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.
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