A hallmark of celiac disease (CeD), a chronic condition driven by cereal gluten exposure, is increase of gut intraepithelial γδ T cells. This may indicate pathogenic involvement of γδ T cells and existence of disease-specific γδ T-cell receptors (TCRs) recognizing defined antigen(s). We performed high-throughput and paired γδ TCR sequencing of single intraepithelial γδ T cells of untreated CeD patients (n = 8; 1821 cells), CeD patients treated with a gluten-free diet (n = 5; 436 cells) and controls (n = 7; 1068 cells). We found that CeD patients, both untreated and treated, had larger and more diverse γδ TCR repertoires, more frequent usage of TRDV1 and TRDV3 and different patterns of TCRγ/TCRδ-pairing compared with controls. Although we observed no public CDR3δ sequences, there were several public CDR3γ sequences—many of which were shared by not only the CeD patients, but also by the controls. These public CDR3s were characterized by few N/P nucleotide insertions with germline and near-germline configuration, hence being easy to generate. Previous findings of CeD-specific CDR3 motifs were not replicated. Thus, being unable to raise evidence for CeD-specific γδ TCRs in this first large, paired γδ TCR single-cell sequencing study, we project challenges for identification of CeD-relevant γδ TCR ligands.

乳糜泻 (CeD) 是一种由谷物麸质暴露引起的慢性疾病，其标志是肠道上皮内 γδ T 细胞增加。这可能表明 γδ T 细胞的致病性参与以及识别特定抗原的疾病特异性 γδ T 细胞受体 (TCR) 的存在。我们对未经治疗的 CeD 患者（n  = 8；1821 个细胞）、接受无麸质饮食治疗的 CeD 患者（n  = 5；436 个细胞）和对照组（n  = 7；1068 个细胞）。我们发现 CeD 患者，无论是未治疗的还是治疗的，都有更大和更多样化的 γδ TCR 库，更频繁地使用TRDV1和TRDV3以及与对照组相比的不同模式的 TCRγ/TCRδ 配对。虽然我们没有观察到公共 CDR3δ 序列，但有几个公共 CDR3γ 序列——其中许多不仅由 CeD 患者共享，而且由对照组共享。这些公共 CDR3 的特点是具有种系和近种系配置的 N/P 核苷酸插入很少，因此易于生成。CeD 特异性 CDR3 基序的先前发现未被复制。因此，由于无法在首次大型配对 γδ TCR 单细胞测序研究中为 CeD 特异性 γδ TCR 提供证据，我们预测了鉴定 CeD 相关 γδ TCR 配体的挑战。