Aggregation of macrophages and fibroblasts is inhibited by a monoclonal antibody to the hyaluronate receptor.
To examine the role of the hyaluronate receptor in cell to cell adhesion, we have employed the K-3 monoclonal antibody (MAb) which specifically binds to the hyaluronate receptor and blocks its ability to interact with hyaluronate. In the first set of experiments, we investigated the spontaneous aggregation of SV-3T3 cells, which involves two distinct mechanisms, one of which is dependent upon the presence of divalent cation and the other is independent. The divalent cation-independent aggregation was found to be completely inhibited by both intact and Fab fragments of the K-3 MAb. In contrast, the K-3 MAb had no effect on the divalent cation-dependent aggregation of cells. In a second set of experiments, we examined alveolar macrophages. The presence of hyaluronate receptors on alveolar macrophages was demonstrated by the fact that detergent extracts of these cells could bind [3H]hyaluronate, and this binding was blocked by the K-3 MAb. Immunoblot analysis of alveolar macrophages showed that the hyaluronate receptor had a Mr of 99,500, which is considerably larger than the 85,000 Mr for that on BHK cells. When hyaluronate was added to suspensions of alveolar macrophages, the cells were induced to aggregate. This effect was inhibited by the K-3 MAb, suggesting that the hyaluronate-induced aggregation was mediated by the receptor.
Green
S J
SJ
Department of Anatomy and Cell Biology, Georgetown University Medical Center, Washington, D.C. 20007.
Tarone
G
G
Underhill
C B
CB
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D002448:Cell Adhesion; D002449:Cell Aggregation; D002460:Cell Line; D002461:Cell Line, Transformed; D002478:Cells, Cultured; D006224:Cricetinae; D005347:Fibroblasts / Q000032:analysis / Q000166:cytology*; D018960:Hyaluronan Receptors; D006820:Hyaluronic Acid / Q000378:metabolism*; D006821:Hyaluronoglucosaminidase / Q000378:metabolism; D015151:Immunoblotting; D008264:Macrophages / Q000032:analysis / Q000166:cytology*; D011650:Pulmonary Alveoli / Q000166:cytology; D011956:Receptors, Cell Surface / Q000276:immunology / Q000378:metabolism*
D000911:Antibodies, Monoclonal; D018960:Hyaluronan Receptors; D011956:Receptors, Cell Surface; D006820:Hyaluronic Acid; D006821:Hyaluronoglucosaminidase