Identifying Regulatory B Cells (B10 Cells) that Produce IL-10 in Mice
Identifying Regulatory B Cells (B10 Cells) that Produce IL-10 in Mice
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DOI：
10.1007/978-1-60761-869-0_7
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发表时间：
2011-01-01
期刊：
影响因子：
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通讯作者：
Tedder, Thomas F.
中科院分区：
文献类型：
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作者：
Matsushita, Takashi;Tedder, Thomas F.
产生IL-10的调节性B细胞现在被认为是一个重要的组成部分。免疫系统。我们在野生型小鼠脾脏中发现了一种罕见的抗原特异性调节性B细胞亚群，具有独特的Ca1d（hi）CD5（+）CD19（hi）表型。我们称这些细胞为B10细胞，因为它们负责大多数B细胞IL-10的产生，它们似乎仅在体外刺激5小时后产生IL-10，并将它们与其他潜在的调节性B细胞亚群区分开来。在脾脏CD1d（hi）CD5（+）CD19（hi）B细胞亚群和其他淋巴组织中也发现了B10祖细胞（B10pro）。本文提供了四种用于鉴定和分离小鼠中产生调节性IL-10的B10细胞的方法。前两种方法用于基于细胞表面表型和胞浆IL-10染色鉴定和计数B10和B10pro细胞。后两种方法用于分离活B10细胞，用于过继转移和功能研究。这些方法将有助于研究B10细胞在炎症、自身免疫性疾病、免疫反应和癌症治疗中的作用。
Regulatory B cells that produce IL-10 are now recognized as an important component. of the immune system. We have identified a rare antigen-specific regulatory B-cell subset with a unique Ca1d(hi)CD5(+)CD19(hi) phenotype in the spleens of wild-type mice. We call these cells B10 cells because they arc responsible for most B cell IL-10 production, they appear to only produce IL-10 after 5 h of in vitro stimulation, and to distinguish them from other potential regulatory B cell subsets. B10 progenitor (B10pro) cells have also been identified within the spleen CD1d(hi)CD5(+)CD19(hi) B-cell subset, and within other lymphoid tissues. Herein, four methods for identifying and isolating regulatory IL-10-producing B10 cells in mice are provided. The first two methods arc used to identify and enumerate B10 and B10pro cells based on their cell surface phenotypes and cytoplasmic IL-10 staining. The last two methods are used to isolate viable B10 cells for adoptive transfer and functional studies. These methods should facilitate the study of B10 cells in inflammation, autoimmune disease, immune responses, and cancer therapy.