Cloning and expression of the panallergen profilin and the major allergen (Ole e 1) from olive tree pollen.
BACKGROUND
Olive tree (Olea europaea) pollen allergy is one of the main causes of allergy in Mediterranean countries and some areas of North America.


OBJECTIVE
To clone olive allergens and to characterize immunologically the purified recombinant allergens.


METHODS
Full-length complementary deoxyribonucleic acid (cDNA) strands encoding olive allergens (Ole e 1) were cloned by polymerase chain reaction amplification and sequenced. Recombinant proteins were produced in Escherichia coli by the use of two different expression systems. Immunoreactivity of the recombinant proteins was tested by ELISA and Western blot with serum from patients with allergy to olive.


RESULTS
Significant sequence polymorphism was found in both allergens. The panallergen profilin was expressed as a nonfusion protein and was purified to homogeneity after a single step of affinity chromatography with a poly-L-proline Sepharose column. One cDNA encoding an Ole e 1 isoform was expressed as a fusion protein consisting of the glutathione S-transferase of Schistosoma japonicum and Ole e 1. The fusion protein was purified to homogeneity by gel filtration chromatography and affinity chromatography with a glutathione-Sepharose column, and digested with thrombin. Both recombinant allergens shared B cell epitopes with the corresponding natural allergens.


CONCLUSIONS
IgE-reactive Ole e 1 and olive profilin expressed in bacteria were purified after simple chromatographic procedures and may be useful for diagnostic purposes.
Asturias
J A
JA
R&D Department, IFIDESA-ARISTEGUI, Bilbao, Spain.
Arilla
M C
MC
Gómez-Bayón
N
N
Martínez
J
J
Martínez
A
A
Palacios
R
R
D000485:Allergens; D000595:Amino Acid Sequence; D052179:Antigens, Plant; D001483:Base Sequence; D003001:Cloning, Molecular; D003285:Contractile Proteins; D018076:DNA, Complementary / Q000235:genetics; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D006801:Humans; D015151:Immunoblotting; D007073:Immunoglobulin E / Q000276:immunology; D008840:Microfilament Proteins / Q000235:genetics* / Q000276:immunology; D008969:Molecular Sequence Data; D010940:Plant Proteins / Q000235:genetics* / Q000276:immunology; D010957:Plasmids; D016133:Polymerase Chain Reaction; D051304:Profilins; D011994:Recombinant Proteins / Q000276:immunology; D006255:Rhinitis, Allergic, Seasonal / Q000276:immunology; D017422:Sequence Analysis, DNA
D000485:Allergens; D052179:Antigens, Plant; D003285:Contractile Proteins; D018076:DNA, Complementary; D008840:Microfilament Proteins; C081946:Ole e I protein, Olea europaea; D010940:Plant Proteins; D051304:Profilins; D011994:Recombinant Proteins; D007073:Immunoglobulin E