Mouse Naive CD4+ T Cell Isolation and In vitro Differentiation into T Cell Subsets
Mouse Naive CD4+ T Cell Isolation and In vitro Differentiation into T Cell Subsets
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DOI：
10.3791/52739
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发表时间：
2015-04-01
影响因子：
1.2
通讯作者：
Reynolds, Joseph M.
中科院分区：
文献类型：
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作者：
Flaherty, Stephanie;Reynolds, Joseph M.
当T细胞受体（TCR）识别MHC II类呈递的同源抗原时，无抗原（naive） CD4(+) T细胞经历扩增和分化为效应亚群。TCR激活时环境中存在的细胞因子信号是决定初始CD4(+) T细胞效应命运的主要因素。尽管初始T细胞激活过程中的细胞因子环境可能是复杂的，并且在体内涉及冗余和相反的信号，但在体外初始CD4(+) T细胞激活过程中添加各种细胞因子组合可以很容易地促进具有标志性细胞因子和转录因子表达的效应T辅助谱系的建立。这种分化实验通常被用作评估被认为促进或抑制某些CD4(+) T辅助亚群发育的靶标的第一步。在分化过程中添加介质，如信号激动剂、拮抗剂或其他细胞因子，也可用于研究特定靶点对T细胞分化的影响。在这里，我们描述了从小鼠中分离初始T细胞的基本方案，以及在体外将初始细胞极化为各种辅助T效应细胞系所需的后续步骤。
Antigen inexperienced (naive) 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 naive CD4(+) T cell. Although the cytokine environment during naive 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 naive T cells from mouse and the subsequent steps necessary for polarizing naive cells to various T helper effector lineages in vitro.