Mouse Naïve CD4+ T Cell Isolation and In vitro Differentiation into T Cell Subsets.
Antigen inexperienced (naïve) CD4(+) T cells undergo expansion and differentiation to effector subsets at the time of T cell receptor (TCR) recognition of cognate antigen presented on MHC class II. The cytokine signals present in the environment at the time of TCR activation are a major factor in determining the effector fate of a naïve CD4(+) T cell. Although the cytokine environment during naïve T cell activation may be complex and involve both redundant and opposing signals in vivo, the addition of various cytokine combinations during naive CD4(+) T cell activation in vitro can readily promote the establishment of effector T helper lineages with hallmark cytokine and transcription factor expression. Such differentiation experiments are commonly used as a first step for the evaluation of targets believed to promote or inhibit the development of certain CD4(+) T helper subsets. The addition of mediators, such as signaling agonists, antagonists, or other cytokines, during the differentiation process can also be used to study the influence of a particular target on T cell differentiation. Here, we describe a basic protocol for the isolation of naïve T cells from mouse and the subsequent steps necessary for polarizing naïve cells to various T helper effector lineages in vitro.
Department of Microbiology and Immunology, The Chicago Medical School, Rosalind Franklin University of Medicine and Science.
Stephanie
S
Flaherty
Department of Microbiology and Immunology, The Chicago Medical School, Rosalind Franklin University of Medicine and Science; joseph.reynolds@rosalindfranklin.edu.
Joseph M
JM
Reynolds
D000818:Animals; D017951:Antigen Presentation; D000941:Antigens / Q000276:immunology; D015496:CD4-Positive T-Lymphocytes / Q000166:cytology* / Q000276:immunology; D002454:Cell Differentiation / Q000276:immunology; D016207:Cytokines / Q000276:immunology; D005260:Female; D008213:Lymphocyte Activation; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D011948:Receptors, Antigen, T-Cell / Q000276:immunology; D016176:T-Lymphocyte Subsets / Q000166:cytology* / Q000276:immunology
D000941:Antigens; D016207:Cytokines; D011948:Receptors, Antigen, T-Cell