Quantification of CD4, CCR5, and CXCR4 levels on lymphocyte subsets, dendritic cells, and differentially conditioned monocyte-derived macrophages
Quantification of CD4, CCR5, and CXCR4 levels on lymphocyte subsets, dendritic cells, and differentially conditioned monocyte-derived macrophages
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DOI：
10.1073/pnas.96.9.5215
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发表时间：
1999-04-27
影响因子：
11.1
通讯作者：
Doms, RW
中科院分区：
文献类型：
--
作者：
Lee, B;Sharron, M;Doms, RW
CCR 5和CXCR 4分别是R5和X4 HIV-1毒株的主要HIV-1辅助受体，并且需要阈值数量的CD 4和趋化因子受体分子来支持病毒感染。因此，我们使用定量荧光激活细胞分选测定法，通过对新鲜全血使用四色荧光激活细胞分选分析来确定各种T细胞系、T细胞亚群、外周血树突状细胞（PBDC）和单核细胞衍生的巨噬细胞上的CD 4、CCR 5和CXCR 4抗体结合位点（ABS）的数量，受体水平在所检查的各种亚群中变化很大，个体之间的变化通常为2至5倍，与CD 4 +/CD 45 RO +/CD 62 L+细胞相比，CCR 5在CD 4 +/CD 45 RO +/CD 62 L-真记忆细胞中的表达水平更高。新鲜的PBDC在检查的白细胞亚群中具有最高数量的CCR 5 ABS，但具有很少的CXCR 4 ABS，提供了用于分选纯化PBDC的策略。PBDC的体外成熟分别导致CCR 5和CXCR 4 ABS的中位数增加3倍和41倍。我们发现，巨噬细胞集落刺激因子引起CCR 5和CXCR 4对巨噬细胞成熟的最大上调（从大约5，000 ABS到大约50，000 ABS），而粒细胞-巨噬细胞集落刺激因子引起CXCR 4的显著降低（从大约5，000 ABS到大约50，000 ABS）。
CCR5 and CXCR4 are the major HIV-1 coreceptors for R5 and X4 HIV-1 strains, respectively, and a threshold number of CD4 and chemokine receptor molecules is required to support virus infection. Therefore, we used a quantitative fluorescence-activated cell sorting assay to determine the number of CD4, CCR5, and CXCR4 antibody-binding sites (ABS) on various T cell lines, T cell subsets, peripheral blood dendritic cells (PBDC), and monocyte-derived macrophages by using four color fluorescence-activated cell sorting analysis on fresh whole blood, Receptor levels varied dramatically among the various subsets examined and typically varied from 2- to 5-fold between individuals, CCR5 was expressed at much higher levels in CD4+/CD45RO+/CD62L-true memory cells compared with CD4+/CD45RO+/CD62L+ cells. Fresh PBDC had the highest number of CCR5 ABS among the leukocyte subsets examined but had few CXCR4 ABS, affording a strategy for sort-purifying PBDC. In vitro maturation of PBDC resulted in median 3- and 41-fold increases in CCR5 and CXCR4 ABS, respectively. We found that macrophage colony-stimulating factor caused the greatest up-regulation of both CCR5 and CXCR4 on macrophage maturation (from approximate to 5,000 to approximate to 50,000 ABS) whereas granulocyte-macrophage colony-stimulating factor caused a marked decrease of CXCR4 (from approximate to 5,000 ABS to