Immunophenotyping of Mouse Colonic Unconventional T Cells by Mapping Cell Phenomics With 22-Color Flow Cytometry Assays.

Lehan Pan, Chunting Shen, Shiyang Huang, Yingchi Yang, Zhongtao Zhang, Dan Tian

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Article in Phenomics (Cham, Switzerland), 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.

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5 · Who and what money

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6 authors.

Lehan PanDepartment of General Surgery, Beijing Friendship Hospital, Capital Medical University, Beijing, 100050 China.ORCID 0000-0003-4679-3379

Chunting ShenBecton Dickinson Medical Devices (Shanghai) Co., Ltd, Beijing, 100025 China.

Shiyang HuangBeijing Key Laboratory of Tolerance Induction and Organ Protection in Transplantation, Beijing, 100032 China.

Yingchi YangDepartment of General Surgery, Beijing Friendship Hospital, Capital Medical University, Beijing, 100050 China.

Zhongtao ZhangDepartment of General Surgery, Beijing Friendship Hospital, Capital Medical University, Beijing, 100050 China.

Dan TianDepartment of General Surgery, Beijing Friendship Hospital, Capital Medical University, Beijing, 100050 China.ORCID 0000-0002-6894-2673

Funding

No grant is acknowledged in the PubMed record.

6 · The paper itself

Abstract

The intestinal epithelium is continually exposed to food-derived antigens and microbiota. This continual exposure requires a delicate immune homeostasis for food tolerance and protection against infection. Unconventional T lymphocytes, including γδT cells, Natural killer T (NKT) cells, Mucosal-Associated Invariant T (MAIT) cells, and Double-Negative T (DNT) cells, typically reside in the mucosal tissue, such as the colon. These cells play crucial roles in maintaining the integrity of the mucosal barrier and immune homeostasis through cytokine secretion and direct cell-mediated effects. Understanding the proportions and functional status of unconventional T lymphocytes in the colon is crucial for elucidating disease mechanisms. In this study, we developed a 22-color flow cytometry panel for comprehensive immunophenotyping of unconventional T lymphocytes in the murine colon. Our optimized protocol included antibody titration and customized gating strategies. We identified distinct populations of unconventional T lymphocytes, including γδT cells, NKT cells and DNT cells, and compared them with conventional T lymphocyte subsets (CD4 Supplementary Information: The online version contains supplementary material available at 10.1007/s43657-025-00237-6.

Indexed as

Activation markersCytokinesImmune checkpoint moleculesImmunophenotypingIntestinal lymphocyteMulti-parametric flow cytometry

Identifiers

PMID41659743

DOI10.1007/s43657-025-00237-6

PMCPMC12881235

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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.