Phenotypic analysis of erythrocytes in sickle cell disease using imaging flow cytometry.
Phenotypic analysis of erythrocytes in sickle cell disease using imaging flow cytometry.
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DOI：
10.1002/cyto.a.24536
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发表时间：
2022-05
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影响因子：
--
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中科院分区：
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--
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镰状细胞病(SCD)患者的红细胞形态和其他表型特征在患者评估中已经分析了几十年。这涉及到多种技术，包括染色血膜的显微分析、流式细胞术和细胞计数。在这里，我们使用成像流式细胞术(IFC)分析SCD血液，这是一项结合了流式细胞术和显微镜的技术，能够同时快速分析细胞形态和细胞表面标记。有了IFC，我们能够自动量化SCD血液中的巨噬细胞。一个重要的多核细胞亚群代表致密细胞，尽管这些细胞不能与其他多核细胞区分开来，除非首先通过密度梯度对血液进行离心。此外，SCD患者的CD71阳性红细胞有两个亚群：一个高表达CD71，呈褶皱状；另一个低表达，呈双凹形态，可能代表分化的后期。部分DAPI和CD49d强阳性的皱褶状红细胞实际上是有核红细胞。与传统的流式细胞术相比，IFC在SCD中发现了更多表达磷脂酰丝氨酸的红细胞，并且这些细胞也可以分为两个亚群。一个群体有弥漫的PS表达，似乎主要由RBC幽灵组成；另一个群体的PS表达总体较低，呈现在Howell-Jolly小体上的密集点状斑点中。这项研究表明，IFC可以快速揭示和量化SCD的RBC特征，这些特征需要许多繁琐的常规方法来识别。因此，IFC很可能是一种评估和监测SCD的有用技术。
The morphology and other phenotypic characteristics of erythrocytes in sickle cell disease (SCD) have been analyzed for decades in patient evaluation. This involves a variety of techniques, including microscopic analysis of stained blood films, flow cytometry, and cell counting. Here, we analyzed SCD blood using imaging flow cytometry (IFC), a technology that combines flow cytometry and microscopy to enable simultaneous rapid-throughput analysis of cellular morphology and cell-surface markers. With IFC, we were able to automate quantification of poikilocytes from SCD blood. An important subpopulation of poikilocytes represented dense cells, although these could not be distinguished from other poikilocytes without first centrifuging the blood through density gradients. In addition, CD71-positive RBCs from SCD patients had two subpopulations: one with high CD71 expression and a puckered morphology and another with lower CD71 expression and biconcave morphology and presumably representing a later stage of differentiation. Some RBCs with puckered morphologies that were strongly positive for DAPI and CD49d were in fact nucleated RBCs. IFC identified more phosphatidylserine-expressing red cells in SCD than did conventional flow cytometry and these could also be divided into two subpopulations. One population had diffuse PS expression and appeared to be composed primarily of RBC ghosts; the other had lower overall PS expression present in intense, punctate dots overlying Howell-Jolly bodies. This study demonstrates that IFC can rapidly reveal and quantify RBC features in SCD that require numerous tedious methods to identify conventionally. Thus, IFC is likely to be a useful technique for evaluating and monitoring SCD.
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影响因子：
64.8
作者：
INGRAM, VM
通讯作者：
INGRAM, VM
影响因子：
20.3
作者：
Mankelow, Tosti J.;Griffiths, Rebecca E.;Anstee, David J.
通讯作者：
Anstee, David J.
影响因子：
20.3
作者：
Gibson, XA;Shartava, A;Goodman, SR
通讯作者：
Goodman, SR
影响因子：
2.2
作者：
Samsel L;McCoy JP Jr
通讯作者：
McCoy JP Jr
DOI：
10.1016/s1383-5718(02)00009-8
发表时间：
2002-03-25
影响因子：
1.9
作者：
Dertinger, SD;Torous, DK;Tometsko, CR
通讯作者：
Tometsko, CR