A monoclonal anti-IgE antibody against an epitope (amino acids 367-376) in the CH3 domain inhibits IgE binding to the low affinity IgE receptor (CD23).
We have produced three different mAb specific for human IgE-Fc. Their binding pattern to either heat-denatured IgE or a family of overlapping IgE-derived recombinant peptides and their ability to affect interaction of IgE with its low affinity receptor Fc epsilon R2/CD23 demonstrate that they recognize distinct epitopes on the IgE molecule. All three mAb were able to induce basophil degranulation as measured by the induction of histamine release. mAb 173 recognizes a thermolabile epitope in the CH4 domain. It does not affect the binding of IgE to Fc epsilon R2/CD23. mAb 272 recognizes a thermostable epitope that maps to a sequence of 36 amino acids (AA) spanning part of the CH2 and CH3 domain and it does not affect the binding of IgE to Fc epsilon R2/CD23. mAb 27 recognizes a thermolabile epitope located on a 10 AA stretch (AA 367-376) in the CH3 domain. This area contains one N-linked oligosaccharide (Asn-371), but the antibody is not directed against carbohydrate because it binds to Escherichia coli-derived IgE peptides. mAb 27 inhibits the binding of IgE to Fc epsilon R2/CD23 but is still capable of reacting with IgE already bound to Fc epsilon R2/CD23. These data suggest that upon binding to Fc epsilon R2/CD23, the IgE molecule engages one of two equivalent-binding sites close to the glycosylated area of the CH3 domain.
Chrétien
I
I
UNICET, Laboratory for Immunological Research, Dardilly, France.
Helm
B A
BA
Marsh
P J
PJ
Padlan
E A
EA
Wijdenes
J
J
Banchereau
J
J
D000818:Animals; D000888:Antibodies, Anti-Idiotypic / Q000502:physiology*; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000502:physiology*; D000918:Antibody Specificity; D000937:Antigen-Antibody Reactions; D000944:Antigens, Differentiation, B-Lymphocyte / Q000276:immunology / Q000378:metabolism*; D001667:Binding, Competitive; D000939:Epitopes / Q000276:immunology*; D006801:Humans; D007073:Immunoglobulin E / Q000276:immunology* / Q000378:metabolism* / Q000502:physiology; D051379:Mice; D008807:Mice, Inbred BALB C; D010446:Peptide Fragments / Q000276:immunology*; D010449:Peptide Mapping; D011961:Receptors, Fc / Q000276:immunology / Q000378:metabolism*; D017455:Receptors, IgE; D011994:Recombinant Proteins / Q000276:immunology; D013329:Structure-Activity Relationship
D000888:Antibodies, Anti-Idiotypic; D000911:Antibodies, Monoclonal; D000944:Antigens, Differentiation, B-Lymphocyte; D000939:Epitopes; D010446:Peptide Fragments; D011961:Receptors, Fc; D017455:Receptors, IgE; D011994:Recombinant Proteins; C026408:anti-IgE antibodies; D007073:Immunoglobulin E