A Monoclonal Antibody Directed Against Activation Antigen on Alveolar Macrophages.

为了阐明肺巨噬细胞的免疫学作用，我们制备了抗兔活化肺泡巨噬细胞的单克隆抗体。用卡介苗（BCG）静脉注射致敏兔的支气管肺泡灌洗（BAL）细胞免疫BALB/c小鼠。从这些小鼠制备的脾细胞与NS-1浆细胞瘤细胞系使用杂交瘤技术融合，我们提出了一个单克隆抗体称为AM-1。采用细胞放射免疫分析、流式细胞术分析和免疫组织化学方法对该单克隆抗体进行筛选和表征。AM-1与正常兔血单核细胞和BAL-M-巨噬细胞均不反应，而与BCG免疫兔BAL-M-巨噬细胞反应呈阳性。接种后7天，来自兔的BAL-macrophages的阳性细胞百分比约为90%。随着BCG致敏时间的延长，阳性率逐渐下降。用Ouchterlony法鉴定AM-1的免疫球蛋白亚类为IgG 2b。有趣的是，用10%胎牛血清或巨噬细胞活化因子孵育可引起肺泡巨噬细胞对AM-1的反应性增加，而用AM-1预处理肺泡巨噬细胞可导致吞噬能力显著降低。并且还表明抗AM-1抗原的表达似乎涉及某些巨噬细胞功能。

We produced a monoclonal antibody against rabbit activated alveolar macrophage, in order to clarify the immunological roles of lung macrophages. BALB/c mice were immunized with bronchoalveolar lavage (BAL) cells obtained from a rabbit sensitized by heat-killed BCG intravenous injection. Spleen cells prepared from these mice were fused with the NS-1 plasmacytoma cell line using hybridoma technology, and we raised a monoclonal antibody called AM-1. The screening and characterization of this monoclonal antibody were carried out employing cellular radioimmunoassay, flow cytometric analysis, and immunohistochemical methods. AM-1 did not react with either blood monocytes or BAL-macrophages obtained from normal rabbits, but was positive for BAL-macrophages from BCG vaccinated rabbits. The percentage of positive cells for BAL-macrophages from rabbits at 7 days after vaccination was about 90%. The positive rate gradually decreased in relation to the time since BCG sensitization. The immunoglobulin subclass of AM-1 was IgG2b checked by Ouchterlony's method. Interestingly, incubation with either 10% fetal calf serum or macrophage activating factor caused increases in AM-1 reactivity of alveolar macrophages, and pretreatment of alveolar macrophages with AM-1 resulted in significantly reduced phagocytic capacity.These results indicates that expression of surface anti AM-1 antigen is associated with an early activation process of alveolar macrophages, and also suggests that expression of anti AM-1 antigen appears to be involved in some of the macrophage functions.