Novel Strategy for Phenotypic Characterization of Human B Lymphocytes from Precursors to Effector Cells by Flow Cytometry.
Novel Strategy for Phenotypic Characterization of Human B Lymphocytes from Precursors to Effector Cells by Flow Cytometry.
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DOI：
10.1371/journal.pone.0162209
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发表时间：
2016
期刊：
影响因子：
3.7
通讯作者：
Jacob MC
中科院分区：
文献类型：
--
作者：
Clavarino G;Delouche N;Vettier C;Laurin D;Pernollet M;Raskovalova T;Cesbron JY;Dumestre-Pérard C;Jacob MC
人类B细胞亚群的精确鉴定和表型特征在基础研究和医学上都是至关重要的。在文献中，对B淋巴细胞表型特征的流式细胞术研究主要集中在描述特定的细胞阶段，或者在单一类型的样本中观察到的特定的细胞阶段。在目前的工作中，我们提出了一个由6个抗体(CD38、CD27、CD10、CD19、CD5和CD45)组成的主干和一个有效的门控策略来在一个分析管中识别大量的B细胞亚群，涵盖了从前体到记忆和浆细胞的整个B细胞分化。此外，通过在8种颜色的组合中添加两种抗体，我们的方法允许分析任何感兴趣的B细胞分化过程中细胞表面标记的调制。因此，我们开发了一组由七种8色抗体组合组成的小组，以确定骨髓、外周血、淋巴结和脐带血样本中B细胞亚群的表型特征。除了双参数表示提供的定性信息外，我们还量化了每个已识别的B细胞亚群上的抗原表达，并提出了一系列信息曲线，显示了17个细胞表面标志在B细胞分化过程中的调控。我们通过流式细胞术的方法提供了一种有效的工具来获得B细胞表面标志物表达的定量数据，这种方法相对容易操作，可以应用于常规的探索。
A precise identification and phenotypic characterization of human B-cell subsets is of crucial importance in both basic research and medicine. In the literature, flow cytometry studies for the phenotypic characterization of B-lymphocytes are mainly focused on the description of a particular cell stage, or of specific cell stages observed in a single type of sample. In the present work, we propose a backbone of 6 antibodies (CD38, CD27, CD10, CD19, CD5 and CD45) and an efficient gating strategy to identify, in a single analysis tube, a large number of B-cell subsets covering the whole B-cell differentiation from precursors to memory and plasma cells. Furthermore, by adding two antibodies in an 8-color combination, our approach allows the analysis of the modulation of any cell surface marker of interest along B-cell differentiation. We thus developed a panel of seven 8-colour antibody combinations to phenotypically characterize B-cell subpopulations in bone marrow, peripheral blood, lymph node and cord blood samples. Beyond qualitative information provided by biparametric representations, we also quantified antigen expression on each of the identified B-cell subsets and we proposed a series of informative curves showing the modulation of seventeen cell surface markers along B-cell differentiation. Our approach by flow cytometry provides an efficient tool to obtain quantitative data on B-cell surface markers expression with a relative easy-to-handle technique that can be applied in routine explorations.
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影响因子：
4.4
作者：
Bohnhorst, JO;Bjorgan, MB;Thompson, KM
通讯作者：
Thompson, KM
DOI：
10.1073/pnas.0511094103
发表时间：
2006-02-14
影响因子：
11.1
作者：
Malaspina, A;Moir, S;Fauci, AS
通讯作者：
Fauci, AS
影响因子：
8.6
作者：
Marie-Cardine, Aude;Divay, Florence;Jacquot, Serge
通讯作者：
Jacquot, Serge
影响因子：
3.4
作者：
Arnoulet, Christine;Bene, Marie C.;Lacombe, Francis
通讯作者：
Lacombe, Francis
影响因子：
20.3
作者：
McKenna, RW;Washington, LT;Kroft, SH
通讯作者：
Kroft, SH