Acquisition of beta-glucuronidase activity by deficient fibroblasts during direct contact with lymphoid cells.
Fibroblasts deficient in beta-glucuronidase acquired high levels of this enzyme when they were co-cultured with concanavalin A-stimulated lymphocytes. Acquired enzyme activity, determined using a single-cell cytochemical assay, was directly proportional to the number of lymphocytes added and persisted for several days in fibroblasts maintained at high density. Lymphocytes did not secret significant levels of beta-glucuronidase into their culture medium, and did not release other substances able to induce synthesis of the enzyme by the deficient fibroblasts. Nor did beta-glucuronidase acquisition result from concanavalin A-mediated uptake of enzyme, since alpha-methylmannoside did not reduce acquired activity. Moreover, lymphocytes from various sources, whether unstimulated or activated by a different mitogen, bacterial lipopolysaccharide, were equally effective in promoting the appearance of beta-glucuronidase. Deficient fibroblasts did not acquire beta-glucuronidase by active endocytosis when co-cultured with lymphocytes, since enzyme extracted from lymphocytes was not itself effective in this respect. Furthermore, mannose 6-phosphate, which did inhibit, endocytosis by deficient fibroblasts of exogenous beta-glucuronidase prepared from 3T3 cells, had no effect on enzyme acquisition by fibroblasts during their co-culture with lymphocytes. Conversely, inhibitors of protein synthesis and energy metabolism, which did not interfere with endocytosis of exogenous enzyme, abolished the acquisition of beta-glucuronidase during co-culture. Deficient fibroblasts did not acquire beta-glucuronidase when they were cultured together with lymphocytes but separated from them by Millipore membranes permeable to exogenous enzyme. Thus, although the mechanism of acquisition is still unclear, the present results suggest that beta-glucuronidase is transferred from lymphocytes to deficient fibroblasts by a process in which direct cell-to-cell contact is obligatory.
D000818:Animals; D002448:Cell Adhesion; D002450:Cell Communication; D002478:Cells, Cultured; D003208:Concanavalin A / Q000494:pharmacology; D004705:Endocytosis; D005347:Fibroblasts / Q000201:enzymology*; D005966:Glucuronidase / Q000037:antagonists & inhibitors / Q000378:metabolism*; D006801:Humans; D007958:Leukocyte Count; D008213:Lymphocyte Activation; D008214:Lymphocytes / Q000201:enzymology*; D051379:Mice; D008808:Mice, Inbred CBA; D010587:Phagocytosis / Q000187:drug effects
D003208:Concanavalin A; D005966:Glucuronidase