Characterization of recombinant cat albumin.
BACKGROUND
Indoor allergens derived from animals and mites often contribute to exacerbation of skin manifestations in atopic dermatitis (AD) patients.


OBJECTIVE
To produce and characterize recombinant cat albumin, a cross-reactive animal allergen.


METHODS
A complete cDNA coding for cat albumin was obtained by RT-PCR amplification from cat liver RNA. Recombinant cat albumin was expressed in Escherichia coli as hexahistidine-tagged protein, purified by nickel affinity chromatography and studied for IgE reactivity with sera from cat-allergic patients by ELISA and immunoblotting. Furthermore, CD203c expression of basophils from cat-allergic patients upon exposure to recombinant cat albumin was analysed.


RESULTS
Recombinant cat albumin, a cross-reactive animal allergen sharing most IgE epitopes with its natural counterpart, was produced in E. coli. It was recognized preferentially by IgE from AD patients and elicited IgE-dependent basophil activation in sensitized patients.


CONCLUSIONS
Recombinant cat albumin may be used as a paradigmatic tool to analyse mechanisms of allergen-triggered exacerbation of AD, for diagnostic and, perhaps for therapeutic purposes.
Reininger
R
R
Institute of Medical and Chemical Laboratory Diagnostics, University of Vienna, Vienna, Austria.
Swoboda
I
I
Bohle
B
B
Hauswirth
A W
AW
Valent
P
P
Rumpold
H
H
Valenta
R
R
Spitzauer
S
S
D000418:Albumins / Q000235:genetics* / Q000276:immunology; D000485:Allergens / Q000235:genetics* / Q000276:immunology; D000818:Animals; D015549:Basophil Degranulation Test; D002415:Cats / Q000276:immunology*; D003429:Cross Reactions; D018076:DNA, Complementary / Q000032:analysis; D003876:Dermatitis, Atopic / Q000175:diagnosis / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D015151:Immunoblotting; D007073:Immunoglobulin E; D010727:Phosphoric Diester Hydrolases / Q000032:analysis; D011755:Pyrophosphatases / Q000032:analysis; D011994:Recombinant Proteins / Q000276:immunology / Q000302:isolation & purification; D020133:Reverse Transcriptase Polymerase Chain Reaction
D000418:Albumins; D000485:Allergens; D018076:DNA, Complementary; C427944:ENPP3 protein, human; D011994:Recombinant Proteins; D007073:Immunoglobulin E; D010727:Phosphoric Diester Hydrolases; D011755:Pyrophosphatases