|Budget Amount *help
¥1,900,000 (Direct Cost : ¥1,900,000)
Fiscal Year 1990 : ¥1,000,000 (Direct Cost : ¥1,000,000)
Fiscal Year 1989 : ¥900,000 (Direct Cost : ¥900,000)
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.