Dieckman Tessa, Schreurs Mette, Lindelauf Ciska, Mahfouz Ahmed, Meijer Caroline R, Pigeaud Louise, van Unen Vincent, Bouma Gerd, Koning Frits
Department of Immunology, Leiden University Medical Center, Leiden, the Netherlands.
Department of Gastroenterology and Hepatology, Amsterdam UMC, Vrije Universiteit Amsterdam, Amsterdam Gastroenterology Endocrinology Metabolism, Amsterdam, the Netherlands.
Gastro Hep Adv. 2024 Sep 4;4(1):100545. doi: 10.1016/j.gastha.2024.08.023. eCollection 2025.
Refractory celiac disease type II (RCDII) is characterized by a clonally expanded aberrant cell population in the small intestine. The role of other tissue-resident immune subsets in RCDII is unknown. Here, we characterized CD8 and CD4 T cells in RCDII duodenum at the single-cell level and .
We applied mass cytometry on CD45 duodenal cells derived from intestinal biopsies (n = 23) and blood samples (n = 20) from RCDII patients and controls. Additionally, we analyzed intestinal biopsies from celiac disease (n = 11) and RCDI (n = 2) patients. We performed single-cell RNA-sequencing on CD45 duodenal cells derived from a RCDII patient, immunofluorescence staining for analysis and flow cytometry for phenotyping of RCDII aberrant and CD8 T cells.
Compared to healthy controls, we observed that CD27PD-1 memory CD8αβ cells and CD4 T regulatories (Tregs) were more abundant in RCDII duodenum (CD8 ∗∗0.0029; CD4 ∗∗∗0.0001). The CD27PD-1 memory CD8αβ cells expressed the tissue-resident marker CD69, immunoregulatory markers (, NKG2A, were enriched for activated pathways and displayed cytotoxic gene signatures (. The absence of CD103 accords with their localization in the lamina propria as determined by analysis. The CD25FoxP3CD27CD127 CD4 Tregs expressed and and costimulatory molecules (, and ) and resided in the lamina propria as well. Flow cytometry confirmed the presence of the inhibitory receptor NKG2A on expanded duodenal CD8 T cells and HLA-E, the ligand for NKG2A, on expanded aberrant cells.
RCDII is characterized by the simultaneous presence of an activated CD27PD-1 memory CD8αβ T cell subset and CD4 Tregs, suggesting that checkpoint blockade with anti-NKG2A/PD-1 and/or anticytotoxic T lymphocyte antigen 4 may be an attractive treatment option.
II型难治性乳糜泻（RCDII）的特征是小肠中存在克隆性扩增的异常细胞群。其他组织驻留免疫亚群在RCDII中的作用尚不清楚。在此，我们在单细胞水平上对RCDII十二指肠中的CD8和CD4 T细胞进行了特征分析。
我们对来自RCDII患者和对照的肠道活检组织（n = 23）和血液样本（n = 20）中的CD45十二指肠细胞进行了质谱流式细胞术分析。此外，我们分析了乳糜泻患者（n = 11）和I型难治性乳糜泻（RCDI）患者（n = 2）的肠道活检组织。我们对来自一名RCDII患者的CD45十二指肠细胞进行了单细胞RNA测序，进行了免疫荧光染色以进行分析，并通过流式细胞术对RCDII异常细胞和CD8 T细胞进行表型分析。
与健康对照相比，我们观察到RCDII十二指肠中CD27PD-1记忆性CD8αβ细胞和CD4调节性T细胞（Tregs）更为丰富（CD8 **0.0029；CD4 ***0.0001）。CD27PD-1记忆性CD8αβ细胞表达组织驻留标志物CD69，免疫调节标志物（，NKG2A）富含活化途径并显示细胞毒性基因特征（。缺乏CD103与其在固有层中的定位一致，这是通过分析确定的。CD25FoxP3CD27CD127 CD4 Tregs表达 和 以及共刺激分子（， 和 ），并且也存在于固有层中。流式细胞术证实扩增的十二指肠CD8 T细胞上存在抑制性受体NKG2A，并且扩增的异常细胞上存在NKG2A的配体HLA-E。
RCDII的特征是同时存在活化的CD27PD-1记忆性CD8αβ T细胞亚群和CD4 Tregs，这表明用抗NKG2A/PD-1和/或抗细胞毒性T淋巴细胞抗原4进行检查点阻断可能是一种有吸引力的治疗选择。
