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A Monoclonal Antibody Directed Against Activation Antigen on Alveolar Macrophages.

针对肺泡巨噬细胞激活抗原的单克隆抗体。

基本信息

批准号：
01570427
负责人：
CHIDA Kingo
金额：
$ 1.22万
依托单位：
Second Division, Department of Internal Medicine, Hamamatsu University School of Medicine.
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1989
资助国家：
日本
起止时间：
1989 至 1990
项目状态：
已结题

项目摘要

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.

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抗。用热灭活卡介苗静脉注射致敏的兔支气管肺泡灌洗液(BAL)细胞免疫BALB/c小鼠。用杂交瘤技术将取自这些小鼠的脾细胞与NS-1浆细胞瘤细胞系融合，制备了一种名为AM-1的单抗。采用细胞放射免疫分析、流式细胞仪分析和免疫组织化学方法对该单抗进行了筛选和鉴定。AM-1与正常兔血单核细胞和BAL-巨噬细胞均不反应，但与卡介苗免疫兔的BAL-巨噬细胞呈阳性反应。免疫后7天，兔BAL-巨噬细胞阳性细胞百分率约为90%。随着卡介苗致敏时间的延长，阳性率逐渐降低。Ouchterlony法检测AM-1免疫球蛋白亚类为IgG2b。有趣的是，与10%胎牛血清或巨噬细胞激活因子孵育后，肺泡巨噬细胞的AM-1反应性增强，而AM-1对肺泡巨噬细胞的吞噬能力显著降低。这些结果表明，肺泡巨噬细胞表面抗AM-1抗原的表达与肺泡巨噬细胞的早期激活过程有关，并提示抗AM-1抗原的表达可能参与了某些巨噬细胞的功能。