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.
Department of Immunology, Leiden University Medical Center, Leiden, The Netherlands.
Cell Mol Gastroenterol Hepatol. 2022;14(1):173-192. doi: 10.1016/j.jcmgh.2022.03.005. Epub 2022 Mar 23.
BACKGROUND & AIMS: Refractory celiac disease type II (RCDII) is a rare indolent lymphoma in the small intestine characterized by a clonally expanded intraepithelial intracellular CD3surfaceCD3CD7CD56 aberrant cell population. However, RCDII pathogenesis is ill-defined. Here, we aimed at single-cell characterization of the innate and adaptive immune system in RCDII.
Paired small intestinal and blood samples from 12 RCDII patients and 6 healthy controls were assessed by single-cell mass cytometry with a 39-cell surface marker antibody panel, designed to capture heterogeneity of the innate and adaptive immune system. A second single-cell mass cytometry panel that included transcription factors and immune checkpoints was used for analysis of paired samples from 5 RCDII patients. Single-cell RNA sequencing analysis was performed on duodenal samples from 2 RCDII patients. Finally, we developed a 40-marker imaging mass cytometry antibody panel to evaluate cell-cell interactions in duodenal biopsy specimens of RCDII patients.
We provide evidence for intertumoral and intratumoral cell heterogeneity within the duodenal and peripheral aberrant cell population present in RCDII. Phenotypic discrepancy was observed between peripheral and duodenal aberrant cells. In addition, we observed that part of the aberrant cell population proliferated and observed co-localization of aberrant cells with CD163 antigen-presenting cells (APCs) in situ. In addition, we observed phenotypic discrepancy between peripheral and duodenal aberrant cells.
Novel high-dimensional single-cell technologies show substantial intertumoral and intratumoral heterogeneity in the aberrant cell population in RCDII. This may underlie variability in refractory disease status between patients and responsiveness to therapy, pointing to the need for personalized therapy in RCDII based on patient-specific immune profiles.
难治性乳糜泻 II 型（RCDII）是一种罕见的小肠惰性淋巴瘤，其特征是克隆性扩展的上皮内细胞内 CD3+表面 CD3+CD7+CD56+异常细胞群。然而，RCDII 的发病机制尚不清楚。在这里，我们旨在对 RCDII 中的固有和适应性免疫系统进行单细胞特征分析。
对 12 例 RCDII 患者和 6 例健康对照者的小肠和血液样本进行单细胞质谱流式细胞术分析，使用 39 个细胞表面标志物抗体面板，设计用于捕获固有和适应性免疫系统的异质性。第二个单细胞质谱流式细胞术面板包括转录因子和免疫检查点，用于分析 5 例 RCDII 患者的配对样本。对 2 例 RCDII 患者的十二指肠样本进行单细胞 RNA 测序分析。最后，我们开发了一个 40 标志物成像质谱流式细胞术抗体面板，用于评估 RCDII 患者十二指肠活检标本中的细胞-细胞相互作用。
我们提供了证据表明，在 RCDII 中，十二指肠和外周异常细胞群体中存在肿瘤内和肿瘤间细胞异质性。在 RCDII 中，外周和十二指肠异常细胞之间存在表型差异。此外，我们观察到部分异常细胞群增殖，并观察到异常细胞与 CD163 抗原呈递细胞（APC）原位共定位。此外，我们观察到外周和十二指肠异常细胞之间存在表型差异。
新型高维单细胞技术显示 RCDII 中异常细胞群体存在大量的肿瘤内和肿瘤间异质性。这可能是患者之间难治性疾病状态的变异性和对治疗的反应性的基础，表明需要根据患者特定的免疫谱为 RCDII 制定个性化治疗方案。
