Evaluation of a 12-Color Flow Cytometry Panel to Study Lymphocyte, Monocyte, and Dendritic Cell Subsets in Humans
Evaluation of a 12-Color Flow Cytometry Panel to Study Lymphocyte, Monocyte, and Dendritic Cell Subsets in Humans
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DOI：
10.1002/cyto.a.20859
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发表时间：
2010-05-01
期刊：
影响因子：
3.7
通讯作者：
Williams, Kenneth C.
中科院分区：
文献类型：
--
作者：
Autissier, Patrick;Soulas, Caroline;Williams, Kenneth C.
在传染病（如人类免疫缺陷病毒（HIV））期间，监测人类免疫细胞群体（例如淋巴细胞，单核细胞和树突状细胞）（DCS）的变化至关重要。但是，识别罕见或异质细胞群的困难可能是限制的。例如，为了准确测量直流子集，八个流式细胞仪参数是理想的。这项工作的目的是使用单个12色流式细胞仪面板分析人类淋巴细胞，单核细胞和DC亚群的表型。红细胞裂解后，将健康人类志愿者的血液洗涤并用12种抗体的鸡尾酒标记。在配备三个激光器的Becton-Dickinson Facsaria（TM）上分析了样品。使用每个血统的5-8 AB组合将数据与谱系特异性面板进行比较。使用FlowJo软件分析获得的数据。我们的12色面板允许在单个管中鉴定以下循环细胞的主要子集：CD4+和CD8+ T淋巴细胞，B淋巴细胞，NK细胞，NKT细胞，单核细胞，单核细胞亚群（CD14和/或CD16），以及五个非过覆盖的非覆盖膜子集HLA-DR+ LIN-亚群：CD34+造血干细胞，CD123+浆细胞类动物DC和三个表达CD16，CD1C（BDCA-1）或CD141或CD141（BDCA-3）的CD11C+髓样DC的三个子集（BDCA-3）。我们已经开发了一个流式细胞仪面板，该面板允许同时检测淋巴细胞和单核细胞群和所有已知的DC亚群。在一个单一面板中研究这些免疫系统的这些主要参与者可能会使我们对HIV感染期间的免疫反应有了更广泛的了解，以及更好地定义单个细胞类型在获得性免疫缺乏综合征（AIDS）发病机理中的作用的能力。 （c）2010年国际细胞仪协会
Monitoring changes in human immune cell populations such as lymphocytes, monocytes, and dendritic cells (DCs) during infectious diseases like human immunodeficiency virus (HIV) is crucial. However, difficulties to identify rare or heterogeneous cell populations can be limiting. For example, to accurately measure DC subsets, eight flow cytometry parameters are ideal. The aim of this work was to analyze the phenotype of human lymphocyte, monocyte, and DC subsets using a single 12-color flow cytometry panel. After erythrocyte lysis, blood from healthy human volunteers was washed and labeled with a cocktail of 12 antibodies. Samples were analyzed on a Becton-Dickinson FACSAria (TM) equipped with three lasers. Data were compared with lineage-specific panels using 5-8 Ab combinations per lineage. Acquired data were analyzed using FlowJo software. Our 12-color panel allows for the identification of the following major subsets of circulating cells in a single tube: CD4+ and CD8+ T lymphocytes, B lymphocytes, NK cells, NKT cells, monocyte subsets (CD14 and/or CD16), and five nonoverlapping HLA-DR+Lin- subsets: CD34+ hematopoietic stem cells, CD123+ plasmacytoid DC, and three subsets of CD11c+ myeloid DC expressing either CD16, CD1c (BDCA-1), or CD141 (BDCA-3). We have developed a single flow cytometry panel that allows for simultaneous detection of the lymphocyte and monocyte cell populations and all known DC subsets. Studying these major players of the immune system in one single panel may give us a broader view of the immune response during HIV infection and the ability to better define the role of individual cell types in Acquired Immune Deficiency Syndrome (AIDS) pathogenesis. (C) 2010 International Society for Advancement of Cytometry