Production of a chimeric allergen derived from the major allergen group 1 of house dust mite species in Nicotiana benthamiana.
Plants are widely accepted as a general platform for the large-scale production of recombinant proteins, which has been demonstrated by the successful expression of various exogenous proteins. Using plants as a bioreactor for mass production of target proteins for vaccines is thought to show the most potential. This study explores whether a chimeric allergen R8, derived from the major allergen group 1 of house dust mites species (Dermatophagoides farinae and Dermatophagoides pteronyssinus), is expressed in tobacco. The highly efficient and useful Tobacco mosaic virus RNA-based overexpression (TRBO) vector was used to investigate expression of the R8 molecule in tobacco by agroinfection. Presence of R8 was detected using SDS-PAGE and Western blotting. Purified allergens were characterized using IgE-binding activity assay and allergen-specific immunotherapy (ASIT) in murine asthmatic models. The recombinant R8 was successfully expressed in tobacco leaves. The pro-peptide was observed in the herbaceous leaf extracts. This protein exhibits properties similar to the parental allergen ProDer f 1 expressed in Escherichia coli or tobacco with respect to IgE immunoreactivity. R8 also rectifies imbalance of TH1/TH2 cells. An herbaceous plant expression system model allows mass production of R8, which might be used in the future for diagnosis of asthma or production of a candidate vaccine for allergen-specific immunotherapy of asthma.
Department of Medical Parasitology, Wannan Medical College, Wuhu, Anhui, China. cpli001@126.com
Chaopin
C
Li
Yuxin
Y
Jiang
Wei
W
Guo
Zhiming
Z
Liu
D000485:Allergens / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D000818:Animals; D000906:Antibodies / Q000097:blood / Q000276:immunology; D039741:Antigens, Dermatophagoides / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D060829:Arthropod Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D001249:Asthma / Q000097:blood / Q000276:immunology / Q000628:therapy; D015153:Blotting, Western; D001992:Bronchoalveolar Lavage Fluid / Q000737:chemistry / Q000276:immunology; D016207:Cytokines / Q000276:immunology / Q000378:metabolism; D040001:Dermatophagoides farinae / Q000235:genetics / Q000276:immunology; D040002:Dermatophagoides pteronyssinus / Q000235:genetics / Q000276:immunology; D003888:Desensitization, Immunologic / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D005822:Genetic Vectors / Q000235:genetics; D007073:Immunoglobulin E / Q000235:genetics / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D011485:Protein Binding / Q000276:immunology; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D014026:Nicotiana / Q000235:genetics / Q000378:metabolism*; D014027:Tobacco Mosaic Virus / Q000235:genetics
D000485:Allergens; D000906:Antibodies; D039741:Antigens, Dermatophagoides; D060829:Arthropod Proteins; D016207:Cytokines; D011993:Recombinant Fusion Proteins; D007073:Immunoglobulin E