Recombinant carp parvalbumin, the major cross-reactive fish allergen: a tool for diagnosis and therapy of fish allergy.
IgE-mediated reactions to fish allergens represent one of the most frequent causes of food allergy. We have constructed an expression cDNA library from carp (Cyprinus carpio) muscle in phage lambda gt11 and used serum IgE from a fish allergic patient to isolate 33 cDNA clones that coded for two parvalbumin isoforms (Cyp c 1.01 and Cyp c 1.02) with comparable IgE binding capacities. Both isoforms represented calcium-binding proteins that belonged to the beta-lineage of parvalbumins. The Cyp c 1.01 cDNA was overexpressed in Escherichia coli, and rCyp c 1.01 was purified to homogeneity. Circular dichroism analysis and mass spectroscopy showed that rCyp c 1.01 represented a folded protein with mainly alpha-helical secondary structure and a molecular mass of 11,416 Da, respectively. rCyp c 1.01 reacted with IgE from all fish-allergic patients tested (n = 60), induced specific and dose-dependent basophil histamine release, and contained most of the IgE epitopes (70%) present in natural allergen extracts from cod, tuna, and salmon. Therefore, it may be used to identify patients suffering from IgE-mediated fish allergy. The therapeutic potential of rCyp c 1.01 is indicated by our findings that rabbit Abs raised against rCyp c 1.01 inhibited the binding of IgE (n = 25) in fish-allergic patients to rCyp c 1.01 between 35 and 97% (84% mean inhibition) and that depletion of calcium strongly reduced IgE recognition of rCyp c 1.01. The latter results suggest that it will be possible to develop strategies for immunotherapy for fish allergy that are based on calcium-free hypoallergenic rCyp c 1.01 derivatives.
Swoboda
Ines
I
Institute of Medical and Chemical Laboratory Diagnostics, University of Vienna, Vienna, Austria.
Bugajska-Schretter
Agnes
A
Verdino
Petra
P
Keller
Walter
W
Sperr
Wolfgang R
WR
Valent
Peter
P
Valenta
Rudolf
R
Spitzauer
Susanne
S
D000485:Allergens / Q000737:chemistry / Q000235:genetics* / Q000276:immunology* / Q000627:therapeutic use; D000595:Amino Acid Sequence; D000818:Animals; D052179:Antigens, Plant; D001483:Base Sequence; D001491:Basophils / Q000276:immunology; D002135:Calcium-Binding Proteins / Q000737:chemistry / Q000235:genetics* / Q000276:immunology*; D002347:Carps / Q000276:immunology*; D003429:Cross Reactions; D018985:Epitopes, B-Lymphocyte / Q000276:immunology; D004926:Escherichia coli / Q000235:genetics; D029941:Fish Proteins; D005512:Food Hypersensitivity / Q000175:diagnosis / Q000276:immunology* / Q000628:therapy; D006636:Histamine Release; D006801:Humans; D007073:Immunoglobulin E / Q000276:immunology; D007074:Immunoglobulin G / Q000276:immunology; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010320:Parvalbumins / Q000737:chemistry / Q000235:genetics* / Q000276:immunology*; D011994:Recombinant Proteins / Q000276:immunology / Q000627:therapeutic use; D017386:Sequence Homology, Amino Acid
D000485:Allergens; D052179:Antigens, Plant; D002135:Calcium-Binding Proteins; C454159:Cyp c 1.01 protein, carp; C454160:Cyp c 1.02 protein, carp; D018985:Epitopes, B-Lymphocyte; D029941:Fish Proteins; D007074:Immunoglobulin G; D010320:Parvalbumins; D011994:Recombinant Proteins; D007073:Immunoglobulin E