Expression of Schistosome-derived Omega-1 with diantennary N-glycans carrying Lewis X motifs in Nicotiana benthamiana plants

R.H.P. Wilbers, L.B. Westerhof, D.R. van Raaij, E. Capuder, A. van der Schuren, D. Nguyen, L. Hussaarts, N. Driessen, C.H. Hokke, M. Yazdanbakhsh, D. Bosch, A. Schots

Research output: Chapter in Book/Report/Conference proceedingAbstract

Abstract

Omega-1 is a T2 ribonuclease (RNase) secreted by Schistosoma mansoni eggs and is a key factor for the induction of Th2 cell differentiation. Induction of Th2 responses by omega-1 is dependent on its RNase activity as well as the N-glycan mediated internalisation by antigen presenting cells. Omega-1 carries two core-difucosylated diantennary N-glycans containing terminal Lewis X motifs. Lewis X is known to be an immunomodulatory glycan due to its interaction with DC-SIGN. However, the main receptor required for omega-1 induced Th2 polarisation was shown to be the mannose receptor. The exact role of N-glycans on the immunomodulatory properties of helminth secreted glycoproteins remains to be elucidated. As the purification of a single glycoprotein from Schistosome egg extracts is relatively inefficient, a platform is required that enables high expression of these helminth glycoproteins with their native N¬-glycan structures. Here we show that Schistosome-derived omega-1 can be efficiently produced in Nicotiana benthamiana plants by means of agroinfiltration. Omega-1 was purified from the intercellular space (apoplast) of Nicotiana benthamiana leafs and was shown to have RNase activity. Omega-1 produced in wild-type plants carried diantennary N-glycans containing typical plant ß1,2-xylose and core a1,3-fucose, but lacked terminal GlcNAc's. By the controlled co-expression of two glycosyltransferases, omega-1 could be engineered to carry terminal Lewis X motifs. Furthermore, both wild-type omega-1 and omega-1 carrying Lewis X motifs were able to induce Th2 cell polarisation. All-in-all our results demonstrate that plants are a promising platform for the expression of helminth glycoproteins carrying engineered N-glycans, which opens up a new field of research.
Original languageEnglish
Title of host publicationProceedings of Molecular and Cellular Biology of Helminths VIII
Pages66-66
Publication statusPublished - 2014
EventMolecular and Cellular Biology of Helminths VIII - Hydra, Greece
Duration: 1 Sep 20146 Sep 2014

Conference

ConferenceMolecular and Cellular Biology of Helminths VIII
CountryGreece
CityHydra
Period1/09/146/09/14

Fingerprint

Tobacco
Polysaccharides
Helminths
Glycoproteins
Th2 Cells
ribonuclease T(2)
Ribonucleases
Glycosyltransferases
Fucose
Schistosoma mansoni
Xylose
Extracellular Space
Antigen-Presenting Cells
Eggs
Ovum
Cell Differentiation
Research

Cite this

Wilbers, R. H. P., Westerhof, L. B., van Raaij, D. R., Capuder, E., van der Schuren, A., Nguyen, D., ... Schots, A. (2014). Expression of Schistosome-derived Omega-1 with diantennary N-glycans carrying Lewis X motifs in Nicotiana benthamiana plants. In Proceedings of Molecular and Cellular Biology of Helminths VIII (pp. 66-66)
Wilbers, R.H.P. ; Westerhof, L.B. ; van Raaij, D.R. ; Capuder, E. ; van der Schuren, A. ; Nguyen, D. ; Hussaarts, L. ; Driessen, N. ; Hokke, C.H. ; Yazdanbakhsh, M. ; Bosch, D. ; Schots, A. / Expression of Schistosome-derived Omega-1 with diantennary N-glycans carrying Lewis X motifs in Nicotiana benthamiana plants. Proceedings of Molecular and Cellular Biology of Helminths VIII. 2014. pp. 66-66
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title = "Expression of Schistosome-derived Omega-1 with diantennary N-glycans carrying Lewis X motifs in Nicotiana benthamiana plants",
abstract = "Omega-1 is a T2 ribonuclease (RNase) secreted by Schistosoma mansoni eggs and is a key factor for the induction of Th2 cell differentiation. Induction of Th2 responses by omega-1 is dependent on its RNase activity as well as the N-glycan mediated internalisation by antigen presenting cells. Omega-1 carries two core-difucosylated diantennary N-glycans containing terminal Lewis X motifs. Lewis X is known to be an immunomodulatory glycan due to its interaction with DC-SIGN. However, the main receptor required for omega-1 induced Th2 polarisation was shown to be the mannose receptor. The exact role of N-glycans on the immunomodulatory properties of helminth secreted glycoproteins remains to be elucidated. As the purification of a single glycoprotein from Schistosome egg extracts is relatively inefficient, a platform is required that enables high expression of these helminth glycoproteins with their native N¬-glycan structures. Here we show that Schistosome-derived omega-1 can be efficiently produced in Nicotiana benthamiana plants by means of agroinfiltration. Omega-1 was purified from the intercellular space (apoplast) of Nicotiana benthamiana leafs and was shown to have RNase activity. Omega-1 produced in wild-type plants carried diantennary N-glycans containing typical plant {\ss}1,2-xylose and core a1,3-fucose, but lacked terminal GlcNAc's. By the controlled co-expression of two glycosyltransferases, omega-1 could be engineered to carry terminal Lewis X motifs. Furthermore, both wild-type omega-1 and omega-1 carrying Lewis X motifs were able to induce Th2 cell polarisation. All-in-all our results demonstrate that plants are a promising platform for the expression of helminth glycoproteins carrying engineered N-glycans, which opens up a new field of research.",
author = "R.H.P. Wilbers and L.B. Westerhof and {van Raaij}, D.R. and E. Capuder and {van der Schuren}, A. and D. Nguyen and L. Hussaarts and N. Driessen and C.H. Hokke and M. Yazdanbakhsh and D. Bosch and A. Schots",
year = "2014",
language = "English",
pages = "66--66",
booktitle = "Proceedings of Molecular and Cellular Biology of Helminths VIII",

}

Wilbers, RHP, Westerhof, LB, van Raaij, DR, Capuder, E, van der Schuren, A, Nguyen, D, Hussaarts, L, Driessen, N, Hokke, CH, Yazdanbakhsh, M, Bosch, D & Schots, A 2014, Expression of Schistosome-derived Omega-1 with diantennary N-glycans carrying Lewis X motifs in Nicotiana benthamiana plants. in Proceedings of Molecular and Cellular Biology of Helminths VIII. pp. 66-66, Molecular and Cellular Biology of Helminths VIII, Hydra, Greece, 1/09/14.

Expression of Schistosome-derived Omega-1 with diantennary N-glycans carrying Lewis X motifs in Nicotiana benthamiana plants. / Wilbers, R.H.P.; Westerhof, L.B.; van Raaij, D.R.; Capuder, E.; van der Schuren, A.; Nguyen, D.; Hussaarts, L.; Driessen, N.; Hokke, C.H.; Yazdanbakhsh, M.; Bosch, D.; Schots, A.

Proceedings of Molecular and Cellular Biology of Helminths VIII. 2014. p. 66-66.

Research output: Chapter in Book/Report/Conference proceedingAbstract

TY - CHAP

T1 - Expression of Schistosome-derived Omega-1 with diantennary N-glycans carrying Lewis X motifs in Nicotiana benthamiana plants

AU - Wilbers, R.H.P.

AU - Westerhof, L.B.

AU - van Raaij, D.R.

AU - Capuder, E.

AU - van der Schuren, A.

AU - Nguyen, D.

AU - Hussaarts, L.

AU - Driessen, N.

AU - Hokke, C.H.

AU - Yazdanbakhsh, M.

AU - Bosch, D.

AU - Schots, A.

PY - 2014

Y1 - 2014

N2 - Omega-1 is a T2 ribonuclease (RNase) secreted by Schistosoma mansoni eggs and is a key factor for the induction of Th2 cell differentiation. Induction of Th2 responses by omega-1 is dependent on its RNase activity as well as the N-glycan mediated internalisation by antigen presenting cells. Omega-1 carries two core-difucosylated diantennary N-glycans containing terminal Lewis X motifs. Lewis X is known to be an immunomodulatory glycan due to its interaction with DC-SIGN. However, the main receptor required for omega-1 induced Th2 polarisation was shown to be the mannose receptor. The exact role of N-glycans on the immunomodulatory properties of helminth secreted glycoproteins remains to be elucidated. As the purification of a single glycoprotein from Schistosome egg extracts is relatively inefficient, a platform is required that enables high expression of these helminth glycoproteins with their native N¬-glycan structures. Here we show that Schistosome-derived omega-1 can be efficiently produced in Nicotiana benthamiana plants by means of agroinfiltration. Omega-1 was purified from the intercellular space (apoplast) of Nicotiana benthamiana leafs and was shown to have RNase activity. Omega-1 produced in wild-type plants carried diantennary N-glycans containing typical plant ß1,2-xylose and core a1,3-fucose, but lacked terminal GlcNAc's. By the controlled co-expression of two glycosyltransferases, omega-1 could be engineered to carry terminal Lewis X motifs. Furthermore, both wild-type omega-1 and omega-1 carrying Lewis X motifs were able to induce Th2 cell polarisation. All-in-all our results demonstrate that plants are a promising platform for the expression of helminth glycoproteins carrying engineered N-glycans, which opens up a new field of research.

AB - Omega-1 is a T2 ribonuclease (RNase) secreted by Schistosoma mansoni eggs and is a key factor for the induction of Th2 cell differentiation. Induction of Th2 responses by omega-1 is dependent on its RNase activity as well as the N-glycan mediated internalisation by antigen presenting cells. Omega-1 carries two core-difucosylated diantennary N-glycans containing terminal Lewis X motifs. Lewis X is known to be an immunomodulatory glycan due to its interaction with DC-SIGN. However, the main receptor required for omega-1 induced Th2 polarisation was shown to be the mannose receptor. The exact role of N-glycans on the immunomodulatory properties of helminth secreted glycoproteins remains to be elucidated. As the purification of a single glycoprotein from Schistosome egg extracts is relatively inefficient, a platform is required that enables high expression of these helminth glycoproteins with their native N¬-glycan structures. Here we show that Schistosome-derived omega-1 can be efficiently produced in Nicotiana benthamiana plants by means of agroinfiltration. Omega-1 was purified from the intercellular space (apoplast) of Nicotiana benthamiana leafs and was shown to have RNase activity. Omega-1 produced in wild-type plants carried diantennary N-glycans containing typical plant ß1,2-xylose and core a1,3-fucose, but lacked terminal GlcNAc's. By the controlled co-expression of two glycosyltransferases, omega-1 could be engineered to carry terminal Lewis X motifs. Furthermore, both wild-type omega-1 and omega-1 carrying Lewis X motifs were able to induce Th2 cell polarisation. All-in-all our results demonstrate that plants are a promising platform for the expression of helminth glycoproteins carrying engineered N-glycans, which opens up a new field of research.

M3 - Abstract

SP - 66

EP - 66

BT - Proceedings of Molecular and Cellular Biology of Helminths VIII

ER -

Wilbers RHP, Westerhof LB, van Raaij DR, Capuder E, van der Schuren A, Nguyen D et al. Expression of Schistosome-derived Omega-1 with diantennary N-glycans carrying Lewis X motifs in Nicotiana benthamiana plants. In Proceedings of Molecular and Cellular Biology of Helminths VIII. 2014. p. 66-66