Comparison of the antibody response to bee venom phospholipase A2 induced by natural exposure in humans or by immunization in mice.
Two human and twelve murine monoclonal antibodies directed against the main bee venom allergen phospholipase A2 (PLA) were evaluated for their fine specificity of binding to antigen and their ability to inhibit the enzymatic activity of the antigen. Antibodies were induced by natural exposure of beekeepers to bee venom or immunization of mice via different methods. Both human monoclonal antibodies (hmAbs) were previously shown to recognize the native three-dimensional conformation of PLA and are directed against discontinuous epitopes which include lysine residue at position 25 as a contact residue. In contrast, six of the murine monoclonal antibodies (mmAbs) bind to the denatured structure of the protein as determined by enzyme-linked immunosorbent assay. The epitopes recognized are located near the C-terminal end (n = 8), in the centre of the polypeptide (n = 1), near the N-terminal end (n = 1) or include the carbohydrate part (n = 2) of the PLA molecule. The capacity of the antibodies to modify the enzymatic activity was also determined. The hmAbs significantly inhibit the enzyme (70-79%), whereas the mmAbs produced various degrees of inhibition (39-100%). Since the X-ray structure of PLA is known, the epitopes can be visualized in the context of the three-dimensional structure of the antigen. A qualitative correlation was found between the location of epitopes and the inhibition pattern. Strong inhibition was seen with those antibodies that recognize epitopes that lie on the surface of the enzyme that is thought to contact the phospholipid bilayer. The results show that even though both hmAbs and most mmAbs inhibit the enzymatic activity of PLA, the antigen-binding properties of antibodies from different species raised after different routes of immunization differ significantly. Thus, detailed epitope mapping studies using murine antibodies prepared by artificial immunization may have limited value in predicting epitope patterns relevant to an antibody response to allergens in humans naturally exposed to antigen/allergen.
Schneider
T
T
Swiss Institute of Allergy and Asthma Research (SIAF), Davos, Switzerland.
Dudler
T
T
Annand
R R
RR
Gelb
M H
MH
King
T P
TP
Suter
M
M
D000485:Allergens / Q000276:immunology*; D000818:Animals; D000911:Antibodies, Monoclonal / Q000378:metabolism*; D000915:Antibody Affinity; D000917:Antibody Formation; D000937:Antigen-Antibody Reactions; D001514:Bee Venoms / Q000201:enzymology*; D004797:Enzyme-Linked Immunosorbent Assay; D018604:Epitope Mapping; D000939:Epitopes / Q000276:immunology* / Q000378:metabolism; D006801:Humans; D006825:Hybridomas; D051379:Mice; D010741:Phospholipases A / Q000276:immunology* / Q000378:metabolism; D054467:Phospholipases A2; D010743:Phospholipids / Q000276:immunology / Q000378:metabolism
D000485:Allergens; D000911:Antibodies, Monoclonal; D001514:Bee Venoms; D000939:Epitopes; D010743:Phospholipids; D010741:Phospholipases A; D054467:Phospholipases A2