Characterizing CD4 T cell differentiation in mouse small intestine using T cell transfer, lamina propria preparation, and flow cytometry

• Published in: STAR protocols Vol. 4; no. 3; p. 102485
• Main Authors: Duan, Jinzhi, Sun, Yanan, Matute, Juan D., Blumberg, Richard S.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 15-09-2023; Elsevier
• Subjects: Animals; CD4-Positive T-Lymphocytes; Cell Differentiation; Flow Cytometry; Flow Cytometry/Mass Cytometry; Immunology; Intestine, Small; Mice; Mucous Membrane; Protocol; Flow Cytometry/Mass Cytometry; Immunology
• ISSN: 2666-1667; 2666-1667
• DOI: 10.1016/j.xpro.2023.102485

Studying gene function in T cells is crucial for understanding physiology and disease pathogenesis. Here, we provide a protocol to examine the role of specific genes in CD4+ T cell differentiation in the intestine. We describe steps for isolating naïve CD4+ T cells from mouse spleens and transferring them to recipient mice. We detail procedures to isolate lamina propria cells and analyze CD4+ T subsets using flow cytometry. This protocol is useful in the study of mucosal immune functions.
For complete details on the use and execution of this protocol, please refer to Duan et al.1
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•Isolating naïve CD4+ T cells from donor mice and transferring them to recipient mice•Isolating mononuclear cells from intestinal lamina propria•Intracellular and extracellular staining of proteins for flow cytometry•Analyzing subsets of CD4+ T cells by flow cytometry
Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.
Studying gene function in T cells is crucial for understanding physiology and disease pathogenesis. Here, we provide a protocol to examine the role of specific genes in CD4+ T cell differentiation in the intestine. We describe steps for isolating naïve CD4+ T cells from mouse spleens and transferring them to recipient mice. We detail procedures to isolate lamina propria cells and analyze CD4+ T subsets using flow cytometry. This protocol is useful in the study of mucosal immune functions.