OMIP-003: phenotypic analysis of human memory B cells.
OMIP-003: phenotypic analysis of human memory B cells.
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DOI：
10.1002/cyto.a.21112
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发表时间：
2011-11
期刊：
影响因子：
3.7
通讯作者：
Sanz, Inaki
中科院分区：
文献类型：
--
作者：
Wei, Chungwen;Jung, John;Sanz, Inaki
背景目前主要的人类B细胞亚群是通过包括IgD、CD 27、CD 38和CD 24在内的多色流式细胞术方案来定义的，以将主要接受的外周血群体（过渡、幼稚、记忆和浆母细胞亚群）分类。总的来说，通过用4-5种颜色方案中的上述标志物的不同组合对几个样品等分试样进行平行染色来评估其他信息标志物（包括IgM、CD 23、CD 10、CD 21和CD 95，以及趋化因子受体表达）的表达。这些方法具有多个缺点，包括：1）“定义”标记如IgD、CD 27和CD 38正确区分主要群体的能力有限; 2）不能确定单一群体中多种标记的实际共表达，这可能导致扩展表型的错误假设，并且由此阻止新亚群的发现;和3）需要大量的细胞来进行多次染色，这是处理稀有样品时的主要实际限制。结合起来，似乎很明显，有限的使用可用的标记不仅不能区分多个群体内的传统核心子集，但可能会导致错误的功能特性的归属。为了解决这些局限性，我们已经开发了几种用于充分表征人B细胞的荧光检测板。这些锚定板共享七个所谓的锚标记。抗CD 19和CD 3的抗体，沿着Fixable Aqua Dead Cell Stain，可明确识别活CD 19 + CD 3 − B细胞。在同一组中纳入四种发育标志物（IgD、CD 24、CD 27和CD 38），使得比较和整合这些不同的分类方案并提供核心人B细胞亚群的精确鉴定成为可能（2）。
BackgroundMajor human B cell subsets are currently defined by pauci-color flow cytometry protocols that typically include IgD, CD27, CD38, and CD24 to classify the major accepted peripheral blood populations (transitional, naïve, memory and plasmablast subsets). By and large, the expression of other informative markers (including IgM, CD23, CD10, CD21 and CD95, as well as chemokine receptor expression) is assessed by parallel staining of several sample aliquots with different combinations of the aforementioned markers in 4–5 color schemes. These approaches suffer from multiple shortcomings including: 1) the limited ability of the “defining” markers such as IgD, CD27 and CD38 to properly discriminate major populations; 2) the inability to ascertain the actual co-expression of multiple markers in a single population possibly leading to faulty assumptions of extended phenotypes and, by extension, preventing the discovery of new sub-populations; and 3) the need for larger number of cells to perform multiple stainings, a major practical limitation when dealing with rare samples. Combined, it seems obvious that limited use of available markers not only fails to differentiate multiple populations within the conventional core subsets, but could potentially lead to erroneous attribution of functional properties. To address these limitations, we have developed several multicolor panels to fully characterize human B cells. These multicolor panels share seven so-called anchor markers. Antibodies against CD19 and CD3, along with the Fixable Aqua Dead Cell Stain, allow the unambiguous identification of live CD19+ CD3− B cells. The inclusion of four developmental markers (IgD, CD24, CD27 and CD38) in the same panel makes it feasible to compare and integrate these different classification schemes and provide precise identifications of the core human B cell subpopulations (2).
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影响因子：
15.3
作者：
Moir, Susan;Ho, Jason;Fauci, Anthony S.
通讯作者：
Fauci, Anthony S.
影响因子：
15.9
作者：
Cappione, A;Anolik, JH;Sanz, I
通讯作者：
Sanz, I
影响因子：
--
作者：
Jacobi, Annett M.;Reiter, Karin;Doerner, Thomas
通讯作者：
Doerner, Thomas
影响因子：
4.4
作者：
Wei, Chungwen;Anolik, Jennifer;Sanz, Inaki
通讯作者：
Sanz, Inaki