Quantitation of rare memory B cell populations by two independent and complementary approaches
Quantitation of rare memory B cell populations by two independent and complementary approaches
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DOI：
10.1016/j.jim.2006.09.005
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发表时间：
2006-12-20
影响因子：
2.2
通讯作者：
Slifka, Mark K.
中科院分区：
文献类型：
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作者：
Amanna, Ian J.;Slifka, Mark K.
目前用于定量临床样品中记忆B细胞（MBC）的方法是有限的，并且通常不允许在单个测定中检测多种MBC特异性。在这里，我们描述了两个独立的方法，抗原特异性MBC定量。首先，开发了一种灵敏的流式细胞术（FC）测定法，用于同时定量两种原型B细胞抗原，破伤风和白喉。其次，开发了基于ELISA的MBC有限稀释测定（LDA），其提供来自单个血液样品的多达8-12种不同MBC频率的定量分析。交叉验证研究表明，FC测量的MBC数量与LDA获得的频率显著相关（R-2 = 0.92，p = 0.0002）。这两种功能上不同的方法将用于精确定量对简单抗原（例如破伤风和白喉）或复杂抗原（例如牛痘或其他病毒）特异的罕见MBC群体，并且适用于各种模型系统。(c)2006 Elsevier B. V.保留所有权利。
Current methodology for quantitation of memory B cells (MBC) in clinical samples is limited and often does not allow for detection of multiple MBC specificities in a single assay. Here we describe two independent approaches to antigen-specific MBC quantitation. First, a sensitive flow cytometry (FC) assay was developed for simultaneous quantitation of two prototypical B cell antigens, tetanus and diphtheria. Second, an ELISA-based MBC limiting dilution assay (LDA) was developed that provides quantitative analysis of up to 8-12 different MBC frequencies from a single blood sample. Cross-validation studies indicated that MBC numbers measured by FC correlated significantly with the frequencies obtained by LDA (R-2 = 0.92, p = 0.0002). These two functionally distinct approaches will be useful for accurate quantitation of rare MBC populations specific for either simple antigens (e.g. tetanus and diphtheria) or complex antigens (e.g. vaccinia or other viruses), and is amenable for use in a variety of model systems. (c) 2006 Elsevier B.V. All rights reserved.