Effector functions of a monoclonal aglycosylated mouse IgG2a: binding and activation of complement component C1 and interaction with human monocyte Fc receptor

Mol Immunol. 1985 Apr;22(4):407-15. doi: 10.1016/0161-5890(85)90125-7.

Abstract

Aglycosylated monoclonal anti-DNP mouse IgG2a produced in the presence of tunicamycin was compared with the native monoclonal IgG2a with respect to its ability to interact with the first component of complement, C1, and to compete with human IgG for binding to human monocyte Fc receptors. The aglycosylated IgG2a was found to bind subcomponent C1q with an equivalent capacity to the native IgG2a, but the dissociation constant was found to be increased three-fold. When activation of C1 by the glycosylated and aglycosylated IgG2a was compared, the rate of C1 activation by the aglycosylated IgG2a was reduced approximately three-fold. In contrast aglycosylation was accompanied by a large decrease (greater than or equal to 50-fold) in the apparent binding constant of monomeric IgG2a to human monocytes. The data suggest that the aglycosylated IgG2a has a structure which differs in the CH2 domain from the native IgG2a, and that the heterogeneous N-linked oligosaccharides of this monoclonal IgG2a which occur at a conserved position in the CH2 domain play a role in maintaining the integrity of its monocyte-binding site. This lack of monocyte binding may result either from a localized conformational change occurring in a single CH2 domain or from an alteration in the CH2-CH2 cross-domain architecture which is normally structured by a pair of opposing and interacting oligosaccharides. The minimal changes in C1q binding and C1 activation suggest that the oligosaccharides are, at most, indirectly involved in these events.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Monoclonal / immunology*
  • Complement Activation*
  • Complement C1 / metabolism*
  • Electrophoresis, Polyacrylamide Gel
  • Humans
  • Immunoglobulin G / immunology
  • Immunoglobulin G / isolation & purification
  • Immunoglobulin G / metabolism*
  • Mice
  • Mice, Inbred Strains
  • Monocytes / immunology*
  • Oligosaccharides / metabolism*
  • Receptors, Fc / metabolism*
  • Tunicamycin / pharmacology

Substances

  • Antibodies, Monoclonal
  • Complement C1
  • Immunoglobulin G
  • Oligosaccharides
  • Receptors, Fc
  • Tunicamycin