Antibodies against macrophages that overlap in specificity with fibroblasts
Antibodies against macrophages that overlap in specificity with fibroblasts
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DOI：
10.1111/j.1523-1755.2005.00358.x
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发表时间：
2005-06-01
影响因子：
19.6
通讯作者：
Neilson, EG
中科院分区：
文献类型：
--
作者：
Inoue, T;Plieth, D;Neilson, EG
背景成纤维细胞可能被误认为巨噬细胞，因为这两种细胞类型共享与靶向单核细胞/巨噬细胞谱系的流行抗体相关的抗原。随着最近对成纤维细胞特异性蛋白1（FSP 1）的描述，我们重新审视了针对巨噬细胞的抗体的特异性，以系统地确定哪些抗体最能区分纤维化组织中的两种细胞类型。在携带GFP转基因的小鼠中产生组织纤维化，所述GFP转基因在FSP 1启动子的控制下编码绿色荧光蛋白。将来自这些标记组织的单细胞悬浮液提交用于使用针对Mac-1的抗体的流式细胞术。Mac-2、Mac-3、F4/80、CD 68、II类主要组织相容性复合体（MHC）和CD 45，并且使用特异性引物组在分选的细胞池中进行编码上述靶抗原的mRNA的cDNA扩增。纤维化组织也用相同的抗体进行免疫组化染色，并在共聚焦显微镜下进行检查。FSP 1（+）成纤维细胞与每种巨噬细胞标志物之间的比较重叠表明，除一种（F4/80）外，所有抗巨噬细胞抗体（Mac-1、Mac-2、Mac-3、CD 68、MHC II类和CD 45）均识别两种细胞类型。针对F4/80的抗体清楚地区分纤维化组织中的巨噬细胞与FSP 1（+）成纤维细胞，并且是小鼠中的优选抗体。
Background. Fibroblasts can be misidentifted as macrophages because both cell types share antigens that are associated with popular antibodies targeting the monocyte/ macrophage lineage. With the recent description of fibroblast-specific protein 1 (FSP1), we revisited the specificity of antibodies directed against macrophages to determine systematically which antibodies best distinguish both cell types in fibrotic tissues.Methods. Tissue fibrosis was produced in mice carrying the GFP transgene encoding green fluorescent protein under the control of the FSP1 promoter. Single cell suspensions from these marked tissues were submitted for flow cytometry using antibodies against Mac-1. Mac-2, Mac-3, F4/80, CD68, major histocompatibility complex (MHC) class II, and CD45, and cDNA amplification of mRNA encoding the above target antigens was performed using specific primer sets in sorted pools of cells. Fibrotic tissues were also stained by immunohistochemistry with the same antibodies and examined under confocal microscopy.Results. Comparison overlap between FSP1(+) fibroblasts with each of the macrophage markers demonstrated that all antimacrophage antibodies (Mac-1, Mac-2, Mac-3, CD68, MHC class II, and CD45) except one (F4/80) recognize both cell types.Conclusion. Antibodies directed against F4/80 clearly distinguish macrophages from FSP1(+) fibroblasts in fibrotic tissues and is the preferred antibody in mice.