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Macrophage membrane glycoproteins that bind Griffonia simplicifolia I-B4: effect on cytotoxicity and protein
A H Warfel1, D Zucker-Franklin, Z Y Zheng
1Department of Medicine, New York University Medical Center, New York 10016.
Thioglycollate elicited peritoneal (TG-Møs) but not resident peritoneal Møs (R-Møs) were found to bind the lectin Griffonia simplicifolia isotype I-B4 (GSI-B4). This was demonstrated by ultrastructural studies and FACS analyses. Membranes from TG-Møs were isolated, separated on SDS-PAGE, electrotransferred onto nitrocellulose, and exposed to peroxidase-labeled GSI-B4. These procedures revealed two major membrane glycoproteins of molecular weights 180,000 and 94,000 daltons that bound the lectin GSI-B4 which has a specificity for recognizing terminal alpha-galactosyl residues. The presence of these epitopes on the two membrane glycoproteins was further substantiated by the fact that treatment of the membranes with alpha-galactosidase destroyed their capacity to bind GSI-B4 and that alpha-D-galactopyranoside but not N-acetyl-D-glucosamine competitively inhibited GSI-B4 from binding to the glycoproteins. Treatment of TG-Møs with GSI-B4 reduced the capacity of interferon (IFN) and lipopolysaccharide (LPS), or IFN alone, to induce Mø mediated cytotoxicity towards tumor cells by as much as 40%. GSI-B4 also caused alterations in the pattern of biosynthetically 35S-methionine labeled secreted proteins as early as 2 hours after contact with TG-Møs. Out of 35 discernible proteins on fluorograms of SDS-PAGE separated proteins, 5 were down-regulated and 9 were enhanced. It is suggested that the two novel Mø membrane proteins may play a role in regulating the response of Mø subpopulations to their humoral and cellular environments.
Thioglycollate elicited peritoneal (TG-Møs) but not resident peritoneal Møs (R-Møs) were found to bind the lectin Griffonia simplicifolia isotype I-B4 (GSI-B4). This was demonstrated by ultrastructural studies and FACS analyses. Membranes from TG-Møs were isolated, separated on SDS-PAGE, electrotransferred onto nitrocellulose, and exposed to peroxidase-labeled GSI-B4. These procedures revealed two major membrane glycoproteins of molecular weights 180,000 and 94,000 daltons that bound the lectin GSI-B4 which has a specificity for recognizing terminal alpha-galactosyl residues. The presence of these epitopes on the two membrane glycoproteins was further substantiated by the fact that treatment of the membranes with alpha-galactosidase destroyed their capacity to bind GSI-B4 and that alpha-D-galactopyranoside but not N-acetyl-D-glucosamine competitively inhibited GSI-B4 from binding to the glycoproteins. Treatment of TG-Møs with GSI-B4 reduced the capacity of interferon (IFN) and lipopolysaccharide (LPS), or IFN alone, to induce Mø mediated cytotoxicity towards tumor cells by as much as 40%. GSI-B4 also caused alterations in the pattern of biosynthetically 35S-methionine labeled secreted proteins as early as 2 hours after contact with TG-Møs. Out of 35 discernible proteins on fluorograms of SDS-PAGE separated proteins, 5 were down-regulated and 9 were enhanced. It is suggested that the two novel Mø membrane proteins may play a role in regulating the response of Mø subpopulations to their humoral and cellular environments.
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