鉴定 B 细胞 IgD 低（BD）调节性 B 细胞的表面表型和调控活性。
Characterization of the Cell Surface Phenotype and Regulatory Activity of B-Cell IgD Low (BD) Regulatory B Cells.
机构信息
Versiti Blood Research Institute, Milwaukee, WI, USA.
Molecular Biology Department, National Research Centre, Cairo, Egypt.
出版信息
Methods Mol Biol. 2021;2270:217-231. doi: 10.1007/978-1-0716-1237-8_12.
B-cell IgD Low (BD) B cells have been shown to promote immunological tolerance by inducing proliferation of CD4Foxp3 T-regulatory cells (Treg) in a glucocorticoid-induced tumor necrosis factor receptor-related protein ligand (GITRL, Tnfsf18)-dependent manner. BD cells constitute a small subset of splenic B lymphocytes that, in mice, are characterized by the B220IgMCD21CD23CD93IgD cell surface expression profile. In this chapter, we show the flow cytometry gating strategy developed to identify and purify BD. In addition, we describe an in vitro assay and two in vivo assays to assess BD regulatory activity by quantitating Treg expansion/proliferation and indicate how they can be used in mouse models of disease. Collectively, these methods are useful to track and quantitate BD and Treg numbers and assess their regulatory activity in inflammatory disease models.
B 细胞免疫球蛋白 D 低（BD）B 细胞已被证明通过糖皮质激素诱导的肿瘤坏死因子受体相关蛋白配体（GITRL，Tnfsf18）依赖性方式诱导 CD4Foxp3 T 调节细胞（Treg）的增殖来促进免疫耐受。BD 细胞构成了脾 B 淋巴细胞的一小部分亚群，在小鼠中，其特征是 B220IgMCD21CD23CD93IgD 细胞表面表达谱。在本章中，我们展示了开发用于鉴定和纯化 BD 的流式细胞术门控策略。此外，我们描述了一种体外测定和两种体内测定方法，用于通过定量 Treg 扩增/增殖来评估 BD 的调节活性，并指出它们如何在疾病的小鼠模型中使用。总之，这些方法可用于跟踪和定量 BD 和 Treg 的数量，并评估它们在炎症性疾病模型中的调节活性。
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