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研究B细胞亚群对于免疫治疗和功能相关的研究非常重要。免疫缺陷性疾病、淋巴瘤和自身免疫性疾病均显示了B细胞亚群异常和功能异常。了解B细胞功能和监测B细胞亚群比例的升高或阶段，可为这些疾病的免疫反应监测提供有效的工具。这里提供了两个protocol：一种是检测人类记忆性B细胞，一种是检测小鼠调节性B细胞。
调节性B细胞，即“Bregs”已被大量实验模型证明，其通过分泌IL-10维持外周免疫耐受。
记忆性B细胞在免疫系统中起着重要作用，可对之前已接触过的抗原做出识别和快速反应。
各类B细胞的产生过程
各类B细胞的产生过程
人记忆性B细胞和小鼠调节性B细胞的检测步骤
人记忆性B细胞和小鼠调节性B细胞的检测步骤
小鼠调节性B细胞结果分析
小鼠调节性B细胞
人记忆性B细胞结果分析
人记忆性B细胞
＝＝＝原文摘要＝＝＝＝＝＝＝
Memory and Regulatory B Cell Sub-population Detection by Flow Cytometry
Mark Santos, Wenying Zhang, Roberto Renteria, Jason Whalley, and Matthew Hsu
EMD Millipore, 28820 Single Oak Drive, Temecula, CA 92590 USA
Abstract
Investigating B cell sub-populations is an important area of research due to its therapeutic relevance and function in the immune response. Immunodeficiency, lymphoma, and autoimmune diseases show abnormal B cell subpopulation and function. Understanding B cell functions and having the ability to monitor an increase or decrease in B cell subpopulation levels can provide a useful tool for understanding the immunological response to certain diseases. This knowledge can provide opportunities to devise therapeutic strategies to help combat against immunological disorders. Here we propose two methods for the detection of B cell subtypes in both human and murine models: one for detection of human memory B cells and one for identifying mouse regulatory B cells. In these assays, we have developed monoclonal antibodies and their conjugates for multi-color analysis to enable simultaneous detection of these cells.
Introduction
Memory B cells play an integral role in the immune system for their ability to recognize and react quickly to previously encountered antigens and protect the body from many and future infections. Originating from lymphocytes that are developed and are activated in the bone marrow, these cells are committed to respond rapidly when re-exposure to pathogens or other invading substances occur. By using anti-human CD5, anti-human CD19, and antihuman CD27 monoclonal directly conjugated antibodies, we were able to determine memory B cell and CD5 + B cells populations, as well as quantitatively measure the percentage of cells within each population.
Regulatory B cells are a unique subset of small white blood cells which produce a potent cytokine, IL-10, a protein that can inhibit immune responses. B10 cells function to protect host tissues from immunopathology during infections. Breg subsets can significantly influence T-cell activation and inflammatory responses through IL- 10 production. Understanding where B10 cells propogate (normally represent 1-3% of the cell population) can be useful information as B10 cell numbers expand during autoimmunity and acute inflammation. By performing multi-parametric analysis using CD5, CD19, and CD1d antibodies by flow cytometry, we are able to accurately discriminate low frequency and phenotypically unique subset of regulatory B cells
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