Anaphylactic and non-anaphylactic murine IgG1 differ in their ability to bind to mast cells: relevance of proper glycosylation of the molecule.
We have previously shown that murine IgG1 antibodies comprise two functionally distinct types regarding their ability to induce mast cell degranulation. In this work, we identified two IgG1-producing hybridomas, both with the same antigenic specificity (anti-DNP), but different in vivo anaphylactic activities. Whereas one of them secretes the anaphylactic IgG1 antibody, as assessed by passive cutaneous anaphylaxis, the other produces the non-anaphylactic IgG1 molecule. The evaluation of the ability of both types of IgG1 to bind to and activate a mouse mast cell line revealed that the anaphylactic IgG1 has a higher binding capacity and releases more beta-hexosaminidase from mast cells than the non-anaphylactic IgG1. Aglycosylated IgG1 obtained by treatment of the anaphylactic IgG1-producing hybridoma line with an inhibitor of N-glycosylation failed to elicit anaphylaxis. In addition, a goat anti-mouse IgG1 antibody reacted less with this aglycosylated IgG1 than with the glycosylated form. These results suggest that the anaphylactic activity of IgG1 antibodies is closely related to their structural conformation and the proper N-glycosylation of these molecules. Finally, the difference in the anaphylactic property between the two types of IgG1 seems to be primarily due to binding to the mast cell surface.
Faquim-Mauro
Eliana L
EL
Department of Immunology, Institute of Biomedical Sciences, University of São Paulo, São Paulo, SP, Brazil.
Jacysyn
Jacqueline F
JF
Macedo
Mahasti S
MS
D000707:Anaphylaxis; D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D000906:Antibodies / Q000737:chemistry; D000937:Antigen-Antibody Reactions; D002448:Cell Adhesion; D002460:Cell Line; D004306:Dose-Response Relationship, Immunologic; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D005434:Flow Cytometry; D006031:Glycosylation; D007074:Immunoglobulin G / Q000737:chemistry* / Q000378:metabolism; D008407:Mast Cells / Q000378:metabolism*; D051379:Mice; D008807:Mice, Inbred BALB C; D011487:Protein Conformation; D017452:Receptors, IgG / Q000378:metabolism; D012867:Skin / Q000276:immunology; D014415:Tunicamycin / Q000494:pharmacology; D001619:beta-N-Acetylhexosaminidases / Q000378:metabolism
D000900:Anti-Bacterial Agents; D000906:Antibodies; D007074:Immunoglobulin G; D017452:Receptors, IgG; D014415:Tunicamycin; D001619:beta-N-Acetylhexosaminidases