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首页> 外文期刊>Clinical and Experimental Immunology: An Official Journal of the British Society for Immunology >Polyclonal B cell activation for accurate analysis of pre-existing antigen-specific memory B cells
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Polyclonal B cell activation for accurate analysis of pre-existing antigen-specific memory B cells

机译：多克隆B细胞激活可准确分析已存在的抗原特异性记忆B细胞

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摘要

The enzyme-linked immunospot (ELISPOT) assay is a widely used tool for enumeration of antigen-specific memory B cells in several disciplines, such as vaccination, cancer immunotherapy and transplantation. For the accurate estimation of antigen-specific memory B cell frequencies, a well-defined B cell activation protocol is pivotal. In this study, we aimed to characterize a polyclonal B cell activation protocol to facilitate optimal monitoring of antigen-specific memory B cell frequencies. Total, naive and memory B cells were activated polyclonally with an α-CD40 monoclonal antibody, cytosine-phosphate-guanine (CPG) oligodeoxynucleotide (ODN) 2006, interleukin (IL)-2, IL-10 and IL-21. Polyclonal activation of B cells resulted in equal cell death ratios in naive and memory B cells. When tested in an antigen-specific system, immunoglobulin (Ig)G spots were detected only in the memory fraction. There was no change in B cell polyclonality due to in-vitro activation. Our data show that the current polyclonal activation protocol may be used reliably to estimate the frequency of memory B cells in ELISPOT assays.
机译：酶联免疫斑点法（ELISPOT）是在多个学科（例如疫苗接种，癌症免疫疗法和移植）中枚举抗原特异性记忆B细胞的一种广泛使用的工具。为了准确估计抗原特异性记忆B细胞的频率，良好定义的B细胞激活方案至关重要。在这项研究中，我们旨在表征多克隆B细胞激活协议，以促进对抗原特异性记忆B细胞频率的最佳监测。用α-CD40单克隆抗体，胞嘧啶-磷酸-鸟嘌呤（CPG）寡脱氧核苷酸（ODN）2006，白介素（IL）-2，IL-10和IL-21多克隆激活总的，幼稚和记忆B细胞。 B细胞的多克隆激活在幼稚和记忆B细胞中导致相等的细胞死亡率。在抗原特异性系统中测试时，仅在记忆级分中检测到免疫球蛋白（Ig）G斑点。由于体外活化，B细胞多克隆性没有改变。我们的数据表明，当前的多克隆激活协议可以可靠地用于估计ELISPOT分析中记忆B细胞的频率。

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