Mitogenic properties of neuraminidase and galactose oxidase-treated lymphoblastoid cells and plasma membranes for peripheral blood lymphocytes.
Generation of aldehydes on terminal D-galactose or N-acetyl-D-galactosamine residues of cell surface glycoproteins by treatment with neuraminidase and galactose oxidase (NAGO) renders some types of cells mitogenic for T lymphocytes. The cell surface molecules required for the presentation of mitogenic signals by NAGO-treated cells are unknown. We tested the mitogenic properties of NAGO-treated lymphoblastoid cell lines (LCL) and subcellular fractions as an initial step in the isolation and characterization of cell surface molecules required for stimulation. We report here that the NAGO-LCL of B cell lineage were potent stimulators, whereas the NAGO-LCL of T cell lineage were weaker and more variable stimulators of lymphocyte proliferation. T-LCL that were stimulatory in indirect stimulation did not induce a mixed lymphocyte response, whereas the B-LCL were positive in both assays. Aldehyde-bearing plasma membrane-enriched subcellular fractions, depleted of nuclear, cytosolic, and mitochondrial components, were mitogenic, and the stimulatory activity was dose dependent. The ability to induce mitogenesis was abrogated by reduction of cell surface aldehyde groups. The results indicate that lymphocyte activation, induced by NAGO-treated stimulatory cells, is a plasma membrane-associated event and does not require the metabolic activity of intact cells. Furthermore, the aldehyde moiety is required but not sufficient for presentation of mitogenic signals. The LCL provide a suitable and reproducible source for isolation and characterization of stimulatory cell surface structures.
D000447:Aldehydes / Q000378:metabolism; D001402:B-Lymphocytes / Q000276:immunology; D002460:Cell Line; D002462:Cell Membrane / Q000201:enzymology / Q000276:immunology*; D005692:Galactose Oxidase / Q000494:pharmacology*; D006358:Hot Temperature; D006801:Humans; D007700:Kinetics; D008213:Lymphocyte Activation; D008214:Lymphocytes / Q000276:immunology* / Q000378:metabolism; D009439:Neuraminidase / Q000494:pharmacology*; D013347:Subcellular Fractions / Q000201:enzymology / Q000276:immunology / Q000378:metabolism; D013601:T-Lymphocytes / Q000276:immunology; D013936:Thymidine / Q000378:metabolism
D000447:Aldehydes; D005692:Galactose Oxidase; D009439:Neuraminidase; D013936:Thymidine