Witmer-Pack M D, Crowley M T, Inaba K, Steinman R M
Department of Zoology, Faculty of Science, Kyoto University, Japan.
J Cell Sci. 1993 Aug;105 ( Pt 4):965-73. doi: 10.1242/jcs.105.4.965.
The dendritic cell system operates in situ to capture and present antigens in a form that is immunogenic to T cells. It is likely that dendritic cells require endocytic activity in order to process antigens. On the other hand, macrophages are considered to be the principal cells that internalize substrates in situ. We therefore investigated the phenotype of cells that scavenge the indigestible endocytic tracer, colloidal carbon, by phenotyping the endocytic cells with monoclonal antibodies that help distinguish macrophages from dendritic cells. Of some importance was the monoclonal N418, an antibody to the p150/90 leukocyte beta 2 integrin. FACS analyses on isolates from blood, spleen and peritoneal cavity showed that N418 reacts primarily with dendritic cells. N418 also stained dendritic profiles strongly in tissue sections of liver and spleen, but most of the cells that actively endocytosed carbon in both organs showed little or no N418 staining. Likewise, carbon could not be identified in cells that react with M342, which stains intracellular granules of dendritic cells. In contrast, the carbon-labeled cells in both liver and spleen were labeled with antibodies (SER-4, F4/80, FA11) that bind primarily to isolated macrophages. Therefore the clearance of colloidal carbon in situ reflects the scavenging activity of macrophages and not the endocytic activity that underlies the antigen presenting function of dendritic cells.
树突状细胞系统在原位发挥作用,以一种对T细胞具有免疫原性的形式捕获并呈递抗原。树突状细胞可能需要内吞活性来处理抗原。另一方面,巨噬细胞被认为是原位内化底物的主要细胞。因此,我们通过用有助于区分巨噬细胞和树突状细胞的单克隆抗体对吞噬不可消化的内吞示踪剂——胶体碳的细胞进行表型分析,来研究这些细胞的表型。其中具有一定重要性的是单克隆抗体N418,它是一种针对p150/90白细胞β2整合素的抗体。对血液、脾脏和腹腔分离物进行的荧光激活细胞分选分析表明,N418主要与树突状细胞发生反应。N418在肝脏和脾脏的组织切片中也强烈染色树突状细胞形态,但在这两个器官中积极内吞碳的大多数细胞几乎没有或没有N418染色。同样,在与M342反应的细胞中也无法鉴定出碳,M342可染色树突状细胞的细胞内颗粒。相比之下,肝脏和脾脏中被碳标记的细胞用主要与分离的巨噬细胞结合的抗体(SER-4、F4/80、FA11)进行标记。因此,原位胶体碳的清除反映了巨噬细胞的吞噬活性,而不是树突状细胞抗原呈递功能所依赖的内吞活性。