Abstract
Type I allergy is an immunological disorder triggered by allergens and causes significant health problems. The major allergen of birch pollen is Bet v 1, which belongs to the pathogen-related protein 10 (PR-10) family. Here, we established a rapid and robust method for the production of Bet v 1 in Nicotiana benthamiana leaves, with binding activity to allergic patients’ IgE. The Bet v 1 allergen was expressed in N. benthamiana using a strong agroinfiltration-based transient protein expression system, which consists of a deconstructed geminiviral vector system with a double terminator. Five days post-infiltration, the allergen concentration in N. benthamiana leaves was 1.2 mg/g of fresh mass, being this the maximum yield of Bet v 1 in plants reported up to now. A part of plant-derived Bet v 1 was glycosylated. Bet v 1 purified from N. benthamiana or Brevibacillus brevis was used to carry out enzyme-linked immunoassays; both recombinant allergens were found to have comparable binding properties to the IgE of allergic patients. These results suggest that our plant expression system allows rapid and robust production of the allergen, which keeps the immunogenicity.
Highlights
Type I allergy or immediate hypersensitivity is the consequence of the interaction between an allergen and immunoglobulin E (IgE) antibodies, which are attached to mast cells
Bet v 1 expressed in N. benthamiana was purified with Ni+ column and the purified protein was separated by SDS-PAGE and the gel was stained with Coomassie brilliant blue (CBB) (Supplementary Figure S2A)
A dilution series of the purified recombinant Bet v 1 protein from B. brevis was loaded on the same gel (Figure 1B) to estimate the amount of N. benthamianaderived Bet v 1 based on the results obtained with the image analyzer (Figure 1D)
Summary
Type I allergy or immediate hypersensitivity is the consequence of the interaction between an allergen and immunoglobulin E (IgE) antibodies, which are attached to mast cells. It is estimated that 300–400 million people in the world is affected by type 1 allergies, causing 120 billion Euros in health care costs, sick leaves, and economic losses (Calderon et al, 2012). These symptoms are elicited by environmental aeroallergens such as pollen or house dust mites. More than 100 million patients suffer from allergy to birch pollen (Moverare et al, 2005). The major birch pollen allergen is Bet v 1
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