Identification of a Hevea brasiliensis latex manganese superoxide dismutase (Hev b 10) as a cross-reactive allergen.
BACKGROUND
Cross-reactive allergens play an increasingly important role in latex allergy in complicating both the diagnosis and time course of allergic symptoms. Manganese superoxide dismutase (MnSOD), a ubiquitous protein of prokaryotic and eukaryotic organisms, was described as a cross-reactive allergen in Aspergillus fumigatus. Little information is available on the importance of this pan-allergen in Hevea brasiliensis latex. The aim of this study was to clone and express MnSOD from H. brasiliensis latex, and to obtain the soluble and immunologically active recombinant allergen for diagnosis of latex allergy and to investigate possible cross-reactivities with the structurally related A. fumigatus and human MnSODs.


METHODS
A complementary DNA coding for Hevea latex MnSOD was amplified by PCR. The recombinant protein was produced in Escherichia coli with an N-terminal hexahistidyl tag. Enzymatic activity of the recombinant protein was determined using an enzyme assay for SODs. IgE immunoblotting and IgE inhibition assays were performed to characterize the recombinant allergen and its cross-reactivity.


RESULTS
A Hevea latex MnSOD consisting of 206 amino acid residues was cloned and expressed in E. coli. The allergen was designated Hev b 10. The recombinant protein was enzymatically active, indicating the correct folding of the protein. In immunoblots, latex- as well as A. fumigatus-allergic patients revealed IgE binding to recombinant (r)Hev b 10. Cross-reactivity to Asp f 6, the MnSOD from A. fumigatus, and human MnSOD was determined by inhibition of IgE binding to these MnSODs by rHev b 10.


CONCLUSIONS
Hev b 10 is a new cross-reactive allergen of H. brasiliensis which belongs to the 'latex-mold' group of latex allergens. Furthermore, it is a candidate for primary sensitization in patients allergic to the pan-allergen MnSOD.
Wagner
S
S
Department of Pathophysiology, University of Vienna, Vienna, Austria.
Sowka
S
S
Mayer
C
C
Crameri
R
R
Focke
M
M
Kurup
V P
VP
Scheiner
O
O
Breiteneder
H
H
D000485:Allergens / Q000235:genetics / Q000276:immunology*; D000595:Amino Acid Sequence; D000908:Antibodies, Fungal / Q000276:immunology; D000941:Antigens / Q000235:genetics / Q000276:immunology*; D000946:Antigens, Fungal / Q000276:immunology; D052179:Antigens, Plant; D001232:Aspergillus fumigatus / Q000201:enzymology / Q000276:immunology; D003001:Cloning, Molecular; D003429:Cross Reactions; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli; D005260:Female; D005656:Fungal Proteins / Q000276:immunology; D006801:Humans; D007073:Immunoglobulin E / Q000276:immunology; D007840:Latex / Q000737:chemistry*; D020315:Latex Hypersensitivity / Q000276:immunology*; D008297:Male; D008969:Molecular Sequence Data; D010940:Plant Proteins / Q000235:genetics / Q000276:immunology*; D011993:Recombinant Fusion Proteins / Q000276:immunology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D013482:Superoxide Dismutase / Q000235:genetics / Q000276:immunology*; D014197:Trees / Q000235:genetics / Q000276:immunology*
D000485:Allergens; D000908:Antibodies, Fungal; D000941:Antigens; D000946:Antigens, Fungal; D052179:Antigens, Plant; D005656:Fungal Proteins; D007840:Latex; D010940:Plant Proteins; D011993:Recombinant Fusion Proteins; D007073:Immunoglobulin E; C115317:Asp f 6 allergen; C423742:Hev b 10; D013482:Superoxide Dismutase