Rapid production of recombinant allergens in Nicotiana benthamiana and their impact on diagnosis and therapy.
BACKGROUND
Type I allergies are immunological disorders that afflict a quarter of the world's population. Recombinant allergens have improved the diagnosis of allergic diseases and allow the formulation of new therapeutic approaches. Over 50% of all allergens are of plant origin.


OBJECTIVE
We have applied a novel method of overexpressing plant allergens in the tobacco-related species Nicotiana benthamiana.


METHODS
This method is based on the use of a chimeric tobacco mosaic virus that harbors a foreign gene sequence and directs its transcription after the infection of the host plant.


RESULTS
We have expressed the model allergen Bet v 1, the major birch pollen allergen, and two Hevea brasiliensis latex allergens, the spina-bifida-associated allergens Hev b 1 and Hev b 3, in N. benthamiana using such a viral vector. Bet v 1, Hev b 1 and Hev b 3 produced by this method were recognized by patients' IgE suggesting that the plant-produced allergens were properly folded. Nonpurified Bet v 1 expressed in N. benthamiana leaves had the same immunogenicity as purified Bet v 1 expressed in Escherichia coli or natural Bet v 1 when tested in a murine model of type I allergy.


CONCLUSIONS
We conclude that this plant expression system offers a viable alternative to fermentation-based production of allergens in bacteria or yeasts.
Breiteneder
H
H
Department of Pathophysiology, University of Vienna, Vienna, Austria. Heimo.Breiteneder@akh-wien.ac.at
Krebitz
M
M
Wiedermann
U
U
Wagner
B
B
Essl
D
D
Steinkellner
H
H
Turpen
T H
TH
Ebner
C
C
Buck
D
D
Niggemann
B
B
Scheiner
O
O
D000485:Allergens / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology*; D000818:Animals; D052179:Antigens, Plant; D003876:Dermatitis, Atopic / Q000175:diagnosis / Q000628:therapy; D005822:Genetic Vectors; D006801:Humans; D006969:Hypersensitivity, Immediate / Q000175:diagnosis* / Q000628:therapy*; D007073:Immunoglobulin E / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D010940:Plant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D010947:Plants, Toxic; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000276:immunology; D014026:Nicotiana / Q000235:genetics* / Q000821:virology; D014027:Tobacco Mosaic Virus / Q000235:genetics
D000485:Allergens; D052179:Antigens, Plant; C098306:Hev b 1 protein, Hevea brasiliensis; D010940:Plant Proteins; D011994:Recombinant Proteins; C061312:Bet v 1 allergen, Betula; C061477:REF protein, Hevea brasiliensis; D007073:Immunoglobulin E