OMIP-043: Identification of human antibody secreting cell subsets
OMIP-043: Identification of human antibody secreting cell subsets
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DOI：
10.1002/cyto.a.23305
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发表时间：
2018-02-01
期刊：
影响因子：
3.7
通讯作者：
Groves, Christopher J.
中科院分区：
文献类型：
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作者：
Carrell, Jeffrey;Groves, Christopher J.
asc是一种分化的B细胞，是体液免疫的主要贡献者，因为它们合成和分泌高度特异性的抗体，结合和中和外来抗原(4)。这些细胞存在于原发性和继发性淋巴组织以及外周循环中，其表型根据其组织生态位而变化(5)。ASCs在外周血中出现的频率非常低，虽然它们在扁桃体、脾脏和骨髓中更为丰富，但在所有情况下，它们都被认为是罕见的细胞，这使得它们的研究具有挑战性(6)。当使用流式细胞术表征这些细胞时，必须注意收集足够的事件，以满足所需的置信度(7)。很少有已发表的ASC分析方案强调泊松统计对于低变异系数（% CV）测量的充分事件收集(8)。我们的目标是5%的变异系数（CV），这导致定期收集400-2,000个ASC门事件。目前已有许多流式细胞术方法对ASC进行了鉴定和描述，然而，对ASC进行阳性鉴定的最重要的决定性因素是免疫球蛋白的合成和分泌。ASC也表达非常高水平的外酶CD38，并且对tnf家族受体CD27呈阳性(9)。事实上，非常高的CD38表达被认为足以用于ASC的基本鉴定。大规模显示cd38高事件对于ASC识别至关重要，对于未经训练的眼睛来说可能很困难；一旦为CD38检测器建立了细胞仪设置，在此期间将显示事件的数量增加到100,000是有用的
BACKGROUNDASC are differentiated B cells that are the primary contributor to humoral immunity, as they synthesize and secrete highly specific antibodies that bind and neutralize foreign antigens (4). These cells are present in primary and secondary lymphoid tissue as well as the peripheral circulation, and their phenotype is variable depending on their tissue niche (5). ASCs occur at very low frequencies in peripheral blood, and while they are somewhat more abundant in tonsil, spleen, and bone marrow, in all cases they are considered rare cells, making their study a challenge (6). When characterizing these cells using flow cytometry, care must be given to collecting sufficient events that satisfy a desired level of confidence (7). Few published protocols for analysis of ASC have emphasized Poisson statistics for adequate event collection for low percent coefficient of variation (% CV) measurements (8). We targeted a 5% coefficient of variation (CV), which resulted in regularly collecting 400–2,000 events in the ASC gate.ASC have been identified and described using numerous flow cytometry methods, however, the most important defining factor for positive identification of ASC is the synthesis and secretion of immunoglobulin. ASC also express very high levels of the ectoenzyme CD38 and are positive for the TNF-family receptor CD27 (9). Very high CD38 expression is in fact considered adequate for basic identification of ASC. On-scale display of CD38high events is critical for ASC identification, and can be difficult for the untrained eye; once cytometer settings are established for the CD38 detector, it is useful to increase the number of displayed events to 100,000 during