Cloning and expression of a major allergen from Cupressus arizonica pollen, Cup a 3, a PR-5 protein expressed under polluted environment.
BACKGROUND
This paper describes the cloning and expression of the Cupressus arizonica pollen protein Cup a 3. In addition, we present its modulation under polluted environmental conditions. Species of the Cupressaceae family are important because of their high sensitization prevalence.


METHODS
Cup a 3 cloning is based on the sequence of the homologous protein Jun a 3. Cup a 3 was expressed with good yield in the methylotropic yeast Pichia pastoris.


RESULTS
Recombinant Cup a 3 (rCup a 3) contains 199 amino acids, 10 potential phosphorylation sites and no glycosylation sites. By immunoblot 63% of cypress allergic patients had specific immunoglobulin E antibodies against rCup a 3 (n = 104). This major allergen is homologous to members of the pathogenesis-related proteins (PR-5 group) and contributes to the overall allergenicity of C. arizonica pollen. Our results show that the increased expression of Cup a 3 is dependent on the pollution in the area where the pollen has been collected, being higher under polluted conditions.


CONCLUSIONS
Cup a 3 is a PR-5 protein derived from C. arizonica pollen. The expression of the protein under polluted conditions has a direct incidence on the pollen allergenicity, as has been demonstrated by skin tests and Radioallergosorbent test inhibition.
Cortegano
I
I
Immunology Department, Fundación Jiménez Díaz, Madrid, Spain.
Civantos
E
E
Aceituno
E
E
del Moral
A
A
López
E
E
Lombardero
M
M
del Pozo
V
V
Lahoz
C
C
D000328:Adult; D000485:Allergens / Q000235:genetics* / Q000378:metabolism*; D000595:Amino Acid Sequence; D052179:Antigens, Plant; D003001:Cloning, Molecular; D029781:Cupressus / Q000276:immunology*; D004787:Environmental Pollution; D005260:Female; D005784:Gene Amplification; D015870:Gene Expression; D006801:Humans; D006967:Hypersensitivity / Q000276:immunology; D008297:Male; D008969:Molecular Sequence Data; D010843:Pichia / Q000378:metabolism; D011058:Pollen / Q000276:immunology; D011852:Radioallergosorbent Test; D011994:Recombinant Proteins / Q000378:metabolism
D000485:Allergens; D052179:Antigens, Plant; C488366:Cup a 3 allergen, Cupressus arizonica; D011994:Recombinant Proteins