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

Kathrin Elisabeth Paulus, Vera Mahler, Martin Pabst, Karl Heinz Kogel, Friedrich Altmann, Uwe Sonnewald

Research output: Contribution to journalArticleScientificpeer-review

14 Citations (Scopus)

Abstract

Complex plant N-glycans containing (31,2-xylose and core a1,3-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, (31,2-xylosyltransferase (XylT), and core a1,3-fucosyltransferase (FucT) are responsible for the transfer of (31,2-linked xylose and core a1,3-linked fucose residues to N-glycans of glycoproteins, respectively. To test the clinical relevance of (31,2-xylose-containing epitopes, expression of the tomato (31,2-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 (31,2-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 anti-xylose-specifc IgG antibodies revealed a strong reduction of (31,2-xylose-containing epitopes. Incubating protein extracts from untransformed controls and XylT_RNAi plants with sera from tomato allergic patients showed a patient-specifc reduction in IgE-binding, indicating a reduced allergenic potential of XylT_RNAi tomato fruits, in vitro. To elucidate the clinical relevance of (31,2-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 (31,2-xylose-containing epitopes in vivo.

Original languageEnglish
Article number42
JournalFrontiers in Plant Science
Volume2
Issue numberAUG
DOIs
Publication statusPublished - 29 Aug 2011

Keywords

  • 2-xylosyltransferase
  • CCDs
  • N -glycosylation
  • RNA interference
  • Tomato allergy
  • β

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