title: Expression of α-1,6-fucosyltransferase (FUT8) in rice grain and immunogenicity evaluation of plant-specific glycans.
abstract: Rice seed is a cost-effective bioreactor for the large-scale production of pharmaceuticals. However, convincing evidence of the immunogenicity of plant-specific glycans is still limited although plant-specific glycans are considered potential allergic antigens. In the present study, we found that the α-1,3-fucose content of the glycoprotein produced from rice seed was much lower than that in leaf, and conversely, a higher β-1,2-xylose content was detected in seed than that in leaf. We detected the α-1,6-fucose content in the glutelin and recombinant human α1-antitrypsin (OsrAAT). The further results in a line containing AAT and FUT8 genes indicated that the α-1,6-fucose content of modified glycosylated recombinant α1-antitrypsin (mgOsrAAT) was 38.4%, while glutelin was only 6.8%. Interestingly, the α-1,3-fucose content of mgOsrAAT was significantly reduced by 59.8% compared with that of OsrAAT. Furthermore, we assessed the immunogenicity of OsrAAT, mgOsrAAT and human α1-antitrypsin (hAAT) using an animal system. The PCA results indicated no significant differences in the IgG, IgM and IgE titers among OsrAAT, mgOsrAAT and hAAT. Further studies revealed that those antibodies were mainly from α-1,3-fucose, but not from β-1,2-xylose, indicating that α-1,3-fucose was the major immunogenic resource. Our results demonstrated that α-1,3-fucose contents in seed proteins was much less than that of leaf, and could not be a plant-specific glycan because it also exists in human proteins.
authors.affiliation: State Key Laboratory of Hybrid Rice, College of Life Sciences, Wuhan University, Wuhan 430072, China.
authors.forename: Xianghong
authors.initials: X
authors.lastname: Wang
authors.affiliation: State Key Laboratory of Hybrid Rice, College of Life Sciences, Wuhan University, Wuhan 430072, China.
authors.forename: Daiming
authors.initials: D
authors.lastname: Jiang
authors.affiliation: Healthgen Biotechnology Corp., Gaoxin Avenue, Wuhan 430074, China.
authors.forename: Jingni
authors.initials: J
authors.lastname: Shi
authors.affiliation: State Key Laboratory of Hybrid Rice, College of Life Sciences, Wuhan University, Wuhan 430072, China. Electronic address: dyang@whu.edu.cn.
authors.forename: Daichang
authors.initials: D
authors.lastname: Yang
mesh_terms: D000818:Animals; D000906:Antibodies / Q000097:blood; D056625:Endosperm / Q000737:chemistry / Q000201:enzymology / Q000235:genetics / Q000276:immunology; D005643:Fucose / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D005647:Fucosyltransferases / Q000096:biosynthesis* / Q000235:genetics* / Q000378:metabolism; D005983:Glutens; D006023:Glycoproteins / Q000737:chemistry / Q000276:immunology / Q000378:metabolism; D006031:Glycosylation; D006168:Guinea Pigs; D006801:Humans; D008297:Male; D012275:Oryza / Q000737:chemistry / Q000201:enzymology* / Q000235:genetics* / Q000276:immunology; D010940:Plant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D030821:Plants, Genetically Modified; D011134:Polysaccharides / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism; D011817:Rabbits; D014994:Xylose / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D000515:alpha 1-Antitrypsin / Q000235:genetics / Q000276:immunology
chemical_list: D000906:Antibodies; D006023:Glycoproteins; D010940:Plant Proteins; D011134:Polysaccharides; D000515:alpha 1-Antitrypsin; D005643:Fucose; D005983:Glutens; D014994:Xylose; D005647:Fucosyltransferases
keywords: Immunogenicity of plant-specific glycans; Molecular pharming; N-glycosylation; Plant-specific glycans; Rice seed