Tan Likai, Fichtner Alina Suzann, Bruni Elena, Odak Ivan, Sandrock Inga, Bubke Anja, Borchers Alina, Schultze-Florey Christian, Koenecke Christian, Förster Reinhold, Jarek Michael, von Kaisenberg Constantin, Schulz Ansgar, Chu Xiaojing, Zhang Bowen, Li Yang, Panzer Ulf, Krebs Christian F, Ravens Sarina, Prinz Immo
Institute of Immunology, Hannover Medical School (MHH), Hannover, Germany.
Institute of Systems Immunology, Hamburg Center for Translational Immunology (HCTI), University Medical Center Hamburg-Eppendorf, Hamburg, Germany.
Sci Immunol. 2021 Apr 23;6(58). doi: 10.1126/sciimmunol.abf0125.
Accumulating evidence suggests that the mouse embryonic thymus produces distinct waves of innate effector γδ T cells. However, it is unclear whether this process occurs similarly in humans and whether it comprises a dedicated subset of innate-like type 3 effector γδ T cells. Here, we present a protocol for high-throughput sequencing of and pairs that comprise the clonal γδTCR. In combination with single-cell RNA sequencing, multiparameter flow cytometry, and TCR sequencing, we reveal a high heterogeneity of γδ T cells sorted from neonatal and adult blood that correlated with TCR usage. Immature γδ T cell clusters displayed mixed and diverse TCRs, but effector cell types segregated according to the expression of either highly expanded individual Vδ1 TCRs or moderately expanded semi-invariant Vγ9Vδ2 TCRs. The Vγ9Vδ2 T cells shared expression of genes that mark innate-like T cells, including (encoding PLZF), , and , but consisted of distinct clusters with unrelated Vγ9Vδ2 TCR clones characterized either by , , and cytotoxicity-associated gene expression (type 1) or by , , , and expression (type 3). Effector γδ T cells with type 1 and type 3 innate T cell signatures were detected in a public dataset of early embryonic thymus organogenesis. Together, this study suggests that functionally distinct waves of human innate-like effector γδ T cells with semi-invariant Vγ9Vδ2 TCR develop in the early fetal thymus and persist into adulthood.
越来越多的证据表明，小鼠胚胎胸腺会产生不同批次的先天性效应γδ T细胞。然而，目前尚不清楚这一过程在人类中是否以类似方式发生，以及它是否包含一个专门的先天性3型效应γδ T细胞亚群。在这里，我们提出了一种对包含克隆性γδTCR的α和β链进行高通量测序的方案。结合单细胞RNA测序、多参数流式细胞术和TCR测序，我们揭示了从新生儿和成人血液中分选出来的γδ T细胞具有高度异质性，这与TCR的使用情况相关。未成熟的γδ T细胞簇显示出混合且多样的TCR，但效应细胞类型根据高度扩增的单个Vδ1 TCR或中度扩增的半不变Vγ9Vδ2 TCR的表达而分离。Vγ9Vδ2 T细胞共享标记先天性样T细胞的基因表达，包括（编码PLZF）、、和，但由不同的簇组成，这些簇具有不相关的Vγ9Vδ2 TCR克隆，其特征分别为、、和细胞毒性相关基因表达（1型）或、、、和表达（3型）。在早期胚胎胸腺器官发生的公共数据集中检测到具有1型和3型先天性T细胞特征的效应γδ T细胞。总之，这项研究表明，具有半不变Vγ9Vδ2 TCR的功能不同批次的人类先天性样效应γδ T细胞在胎儿早期胸腺中发育，并持续到成年期。
