Interest of two-dimensional electrophoretic analysis for the characterization of the individual sensitization to latex allergens.
OBJECTIVE
Latex allergy is a type 1 hypersensitivity reaction that mainly affects high-risk populations such as health care workers, spina bifida-affected or multiply-operated children. Ten molecules have so far been identified and registered as latex allergens (Hev b 1 to Hev b 10). The aim of the present investigation was to identify the major latex allergens by an individual analysis of the IgE response of latex-allergic patients to latex proteins separated by two-dimensional (2-D) gel electrophoresis.


METHODS
Latex proteins from a sap or a glove extract were separated by 2-D electrophoresis and transferred to a nitrocellulose membrane. Each membrane was incubated with the serum of one latex-allergic patient. The most frequently recognized latex allergens were characterized in sap and glove extracts using monoclonal antibodies or amino acid microsequencing.


RESULTS
The one-dimensional screening of 54 patient sera revealed 4 major bands recognized by IgE. The 2-D analysis of the sensitization to latex allergens allows the identification of allergen isoforms and the characterization of an individual response diversity. Hev b 6.01 was recognized by 88.9% of the patients. Protein spots around 14 kD were recognized by 48.1% of the patients and corresponded to Hev b 6.03 as well as other proteins. A not yet characterized doublet of acidic proteins with molecular masses of 43 and 94 kD was recognized by 20.4% of the sera. Only 5.5% of the sera did not recognize any of these 4 major allergens. Hev b 1 is the main protein from the glove extract but was not constantly found in sap extracts.


CONCLUSIONS
One-dimensional electrophoretic analysis of the allergen is usually not sufficient to characterize the individual specificity of the IgE response to latex allergens. Latex-glove proteins which are allergens can be absent from the sap extracts and the sensitization to these allergens could be underestimated. Individual 2-D analysis of the sensitization to latex allergens is useful to define the best allergen mixture required for diagnosis and needed for individual therapy monitoring.
Institut Pasteur, Allergie et Environnement, Paris, France. chardin@odontologie.univ-paris5.fr
H
H
Chardin
M
M
Raulf-Heimsoth
Z
Z
Chen
H P
HP
Rihs
C
C
Mayer
F X
FX
Desvaux
H
H
Sénéchal
G
G
Peltre
D000485:Allergens / Q000737:chemistry* / Q000235:genetics / Q000276:immunology*; D015180:Electrophoresis, Gel, Two-Dimensional; D017575:Gloves, Protective; D006801:Humans; D007073:Immunoglobulin E / Q000097:blood; D007840:Latex / Q000737:chemistry* / Q000276:immunology*; D020315:Latex Hypersensitivity / Q000276:immunology* / Q000628:therapy; D010940:Plant Proteins / Q000276:immunology; D011994:Recombinant Proteins / Q000737:chemistry / Q000276:immunology
D000485:Allergens; D007840:Latex; D010940:Plant Proteins; D011994:Recombinant Proteins; D007073:Immunoglobulin E