Forssman glycosphingolipid as an immunohistochemical marker for mouse stromal macrophages in hematopoietic foci

The immunohistochemical distribution of Forssman glycosphingolipid (GSL) in mouse hematopoietic tissue was examined by using light and electron microscopic immunoperoxidase methods with a highly specific rabbit anti-Forssman GSL antibody. Bone marrow, splenic red pulp, and thymic macrophages, which are closely associated with hematopoietic cells, were stained by the antibody, whereas hematopoietic cells, circulating cells, alveolar macrophages, Kupffer cells, peritoneal resident macrophages, and macrophages derived from granulocyte- macrophage colony-forming units cultured in the presence of L-cell- conditioned medium were not stained. In addition, thymic cortical epithelial cells, the framework of reticular cells of the cortical and medullary regions of the mesenteric lymph node and periarterial lymphoid sheath of the spleen, and some vessels of the tissues examined were also stained. After phlebotomy, the fine cytoplasmic processes of Forssman-positive splenic red pulp macrophages were distributed extensively throughout the erythroid colonies. On the other hand, after hypertransfusion, these macrophages retracted their processes, became round, and tended to aggregate. These results indicate that Forssman GSL can be used as an immunohistochemical marker for stromal macrophages in the hematopoietic foci of the bone marrow and splenic red pulp.