Department of Biology, Boston College, Chestnut Hill, Massachusetts 02467, USA.
Cytometry A. 2010 May;77(5):410-9. doi: 10.1002/cyto.a.20859.
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 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.
监测人类免疫细胞群体（如淋巴细胞、单核细胞和树突状细胞 (DC)）在人类免疫缺陷病毒 (HIV) 等传染病中的变化至关重要。然而，识别稀有或异质细胞群体的困难可能会受到限制。例如，要准确测量 DC 亚群，理想情况下需要 8 个流式细胞术参数。本工作的目的是使用单个 12 色流式细胞术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，用鸡尾酒 12 种抗体标记健康人志愿者的血液。使用配备三个激光器的 Becton-Dickinson FACSAria 对样品进行分析。使用每个谱系 5-8 个 Ab 组合的谱系特异性面板对数据进行比较。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亚群：CD4+和 CD8+T 淋巴细胞、B 淋巴细胞、NK 细胞、NKT 细胞、单核细胞亚群（CD14 和/或 CD16）和五个非重叠 HLA-DR+Lin-亚群：CD34+造血干细胞、CD123+浆细胞样 DC 和三个表达 CD16、CD1c（BDCA-1）或 CD141（BDCA-3）的 CD11c+髓样 DC 亚群。我们开发了一个单个流式细胞术面板，允许同时检测淋巴细胞和单核细胞群体以及所有已知的 DC 亚群。在一个单一的面板中研究这些免疫系统的主要参与者可能会让我们更全面地了解 HIV 感染期间的免疫反应，并能够更好地定义单个细胞类型在获得性免疫缺陷综合征 (AIDS) 发病机制中的作用。（c）2010 年国际细胞分析学会。
