Correlation of IgE/IgG4 milk epitopes and affinity of milk-specific IgE antibodies with different phenotypes of clinical milk allergy.
BACKGROUND
Results from large-scale epitope mapping with a peptide microarray have been shown to correlate with clinical features of milk allergy.


OBJECTIVE
We sought to assess IgE and IgG4 epitope diversity and IgE affinity in different clinical phenotypes of milk allergy and identify informative epitopes that might be predictive of clinical outcomes of milk allergy.


METHODS
Forty-one subjects were recruited from a larger study on the effects of ingesting heat-denatured milk proteins in subjects with milk allergy. Using food challenges, subjects were characterized as being clinically reactive to all forms of milk (n = 17), being tolerant to heated milk (HM) products (n = 16), or having outgrown their milk allergy (n = 8). Eleven healthy volunteers without milk allergy served as control subjects. A peptide microarray was performed by using the previously published protocol.


RESULTS
Subjects with milk allergy had increased epitope diversity compared with those who outgrew their allergy. HM-tolerant subjects had IgE-binding patterns similar to those who had outgrown their allergy, but IgG4-binding patterns that were more similar to those of the allergic group. Binding to higher numbers of IgE peptides was associated with more severe allergic reactions during challenge. There was no association between IgG4 peptides and clinical features of milk allergy. Using a competitive peptide microarray assay, allergic patients demonstrated a combination of high- and low-affinity IgE binding, whereas HM-tolerant subjects and those who had outgrown their milk allergy had primarily low-affinity binding.


CONCLUSIONS
Greater IgE epitope diversity and higher affinity, as determined by using the peptide microarray, were associated with clinical phenotypes and severity of milk allergy.
Pediatric Allergy and Immunology, Mount Sinai School of Medicine, New York, NY 10029-6574, USA. julie.wang@mssm.edu
Julie
J
Wang
Jing
J
Lin
Ludmilla
L
Bardina
Marina
M
Goldis
Anna
A
Nowak-Wegrzyn
Wayne G
WG
Shreffler
Hugh A
HA
Sampson
D000293:Adolescent; D000485:Allergens / Q000276:immunology; D000595:Amino Acid Sequence; D000915:Antibody Affinity / Q000276:immunology*; D002648:Child; D002675:Child, Preschool; D018604:Epitope Mapping; D000939:Epitopes / Q000276:immunology*; D005260:Female; D006358:Hot Temperature; D006801:Humans; D007073:Immunoglobulin E / Q000276:immunology*; D007074:Immunoglobulin G / Q000276:immunology*; D008297:Male; D016269:Milk Hypersensitivity / Q000276:immunology*; D008894:Milk Proteins / Q000276:immunology; D008969:Molecular Sequence Data; D010641:Phenotype; D040081:Protein Array Analysis
D000485:Allergens; D000939:Epitopes; D007074:Immunoglobulin G; D008894:Milk Proteins; D007073:Immunoglobulin E