Abstract
Hybridoma antibodies were produced by fusing spleen cells from mice immunized with rat alveolar macrophages (AM) with P3 x 63Ag8/653 myeloma cells. Some of the hybridomas were processed to achieve monoclonality. Reactivity was tested in an indirect membrane immunofluorescence assay. One of the monoclonal antibodies, called VEP6 antibody, reacted with rat AM but not with peritoneal or splenic macrophages and not with peripheral blood monocytes of this species. It was also unreactive with cells from thymus, bone marrow, non-adherent spleen cells, granulocytes and erythrocytes, but it reacted with thrombocytes. Corynebacterium parvum activated AM were slightly less reactive then 'resident' AM, as determined in a fluorescence activated cell sorter. The VEP6 antigen was studied in polyacrylamide gel electrophoresis after immunoprecipitation from 125I-surface-labelled-macrophage NP40-lysates. Four specific radioactive bands were seen in the precipitates corresponding to molecular weights of 31, 33, 35 and 37,000. A slurred band was seen in this molecular weight range when immunoprecipitates of surface labelled thrombocytes were analysed. This study confirms the heterogeneity of macrophages by showing that certain antigens may be expressed only on a subpopulation of macrophages.
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