Journalartikel

Silencing β1,2-xylosyltransferase in transgenic tomato fruits reveals xylose as constitutive component of IgE-binding epitopes


AutorenlistePaulus, KE; Mahler, V; Pabst, M; Kogel, KH; Altmann, F; Sonnewald, U

Jahr der Veröffentlichung2011

ZeitschriftFrontiers in Plant Science

Bandnummer2

ISSN1664-462X

Open Access StatusGold

DOI Linkhttps://doi.org/10.3389/fpls.2011.00042

VerlagFrontiers Media


Abstract
Complex plant N-glycans containing beta 1,2-xylose and core alpha 1,3-fucose are regarded as the major class of the so-called "carbohydrate cross-reactive determinants" reactive with IgE antibodies in sera of many allergic patients, but their clinical relevance is still under debate. Plant glycosyltransferases, beta 1,2-xylosyltransferase (XylT), and core alpha 1,3-fucosyltransferase (FucT) are responsible for the transfer of beta 1,2-linked xylose and core alpha 1,3-linked fucose residues to N-glycans of glycoproteins, respectively. To test the clinical relevance of beta 1,2-xylose-containing epitopes, expression of the tomato beta 1,2-xylosyltransferase was down-regulated by RNA interference (RNAi) in transgenic plants. Fruits harvested from these transgenic plants were analyzed for accumulation of XylT mRNA, abundance of beta 1,2-xylose epitopes and their allergenic potential. Based on quantitative real-time PCR analysis XylT mRNA levels were reduced up to 10-fold in independent transgenic lines as compared to untransformed control, whereas no xylosylated N-glycans could be revealed by MS analysis. Immunoblotting using antixylose-specific IgG antibodies revealed a strong reduction of beta 1,2-xylose-containing epitopes. Incubating protein extracts from untransformed controls and XylT_ RNAi plants with sera from tomato allergic patients showed a patient-specific reduction in IgE-binding, indicating a reduced allergenic potential of XylT_ RNAi tomato fruits, in vitro. To elucidate the clinical relevance of beta 1,2-xylose-containing complex N-glycans skin prick tests were performed demonstrating a reduced responsiveness of tomato allergic patients, in vivo. This study provides strong evidence for the clinical relevance of beta 1,2-xylose-containing epitopes in vivo.



Autoren/Herausgeber




Zitierstile

Harvard-ZitierstilPaulus, K., Mahler, V., Pabst, M., Kogel, K., Altmann, F. and Sonnewald, U. (2011) Silencing β1,2-xylosyltransferase in transgenic tomato fruits reveals xylose as constitutive component of IgE-binding epitopes, Frontiers in Plant Science, 2, Article 42. https://doi.org/10.3389/fpls.2011.00042

APA-ZitierstilPaulus, K., Mahler, V., Pabst, M., Kogel, K., Altmann, F., & Sonnewald, U. (2011). Silencing β1,2-xylosyltransferase in transgenic tomato fruits reveals xylose as constitutive component of IgE-binding epitopes. Frontiers in Plant Science. 2, Article 42. https://doi.org/10.3389/fpls.2011.00042



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