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

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

Research output: Chapter in Book/Report/Conference proceedingAbstract

Abstract

Plants are an excellent platform for the heterologous production of (glyco)proteins. The surprising tolerance of plants to glyco-engineering allows the production of many different N-glycans. N-glycans play a key role in the interaction between helminth secreted proteins and the host immune system. The exact role of N-glycans on the immunomodulatory properties of helminth secreted glycoproteins remains to be elucidated. Studying the biological function of helminth secreted glycoproteins is challenging, as the purification of a single glycoprotein from helminths or their eggs is relatively inefficient. Plants are an appropriate platform to express these proteins in large quantities with their native N-glycan structures. Omega-1 is a T2 ribonuclease (RNase) secreted by Schistosoma mansoni eggs and is a key factor for the induction of T-helper 2 (Th2) cell differentiation. Induction of Th2 responses by Omega-1 depends on its RNase activity as well as on N-glycan mediated internalisation by antigen presenting cells. Wild type 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 and the mannose receptor. In this study we efficiently produced Schistosome-derived Omega-1 in Nicotiana benthamiana by means of agroinfiltration. The protein 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 The First Conference of the International Society for Plant Molecular Farming
Pages22-22
Publication statusPublished - 2014
EventThe First Conference of the International Society for Plant Molecular Farming, Berlin-Dahlem, Germany -
Duration: 17 Jun 201419 Jun 2014

Conference

ConferenceThe First Conference of the International Society for Plant Molecular Farming, Berlin-Dahlem, Germany
Period17/06/1419/06/14

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Schistosoma
Nicotiana benthamiana
polysaccharides
helminths
ribonucleases
glycoproteins
proteins
agroinfiltration
glycosyltransferases
fucose
antigen-presenting cells
Schistosoma mansoni
extracellular space
apoplast
mannose
xylose
cell differentiation
immune system
engineering
receptors

Cite this

Westerhof, L. B., Wilbers, R. H. P., van Raaij, D. R., Capuder, E., van der Schuren, A., Nguyen, D., ... Schots, A. (2014). Expression of Schistosome-derived Omega-1 with diantennary glycans carrying Lewis X motifs in Nicotiana benthamiana plants. In Proceedings of The First Conference of the International Society for Plant Molecular Farming (pp. 22-22)
Westerhof, L.B. ; Wilbers, R.H.P. ; van Raaij, D.R. ; Capuder, E. ; van der Schuren, A. ; Nguyen, D. ; Hussaarts, L. ; Driessen, N. ; Hokke, C. ; Yazdanbakhsh, M. ; Bosch, D. ; Schots, A. / Expression of Schistosome-derived Omega-1 with diantennary glycans carrying Lewis X motifs in Nicotiana benthamiana plants. Proceedings of The First Conference of the International Society for Plant Molecular Farming. 2014. pp. 22-22
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title = "Expression of Schistosome-derived Omega-1 with diantennary glycans carrying Lewis X motifs in Nicotiana benthamiana plants",
abstract = "Plants are an excellent platform for the heterologous production of (glyco)proteins. The surprising tolerance of plants to glyco-engineering allows the production of many different N-glycans. N-glycans play a key role in the interaction between helminth secreted proteins and the host immune system. The exact role of N-glycans on the immunomodulatory properties of helminth secreted glycoproteins remains to be elucidated. Studying the biological function of helminth secreted glycoproteins is challenging, as the purification of a single glycoprotein from helminths or their eggs is relatively inefficient. Plants are an appropriate platform to express these proteins in large quantities with their native N-glycan structures. Omega-1 is a T2 ribonuclease (RNase) secreted by Schistosoma mansoni eggs and is a key factor for the induction of T-helper 2 (Th2) cell differentiation. Induction of Th2 responses by Omega-1 depends on its RNase activity as well as on N-glycan mediated internalisation by antigen presenting cells. Wild type 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 and the mannose receptor. In this study we efficiently produced Schistosome-derived Omega-1 in Nicotiana benthamiana by means of agroinfiltration. The protein 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 = "L.B. Westerhof and R.H.P. Wilbers and {van Raaij}, D.R. and E. Capuder and {van der Schuren}, A. and D. Nguyen and L. Hussaarts and N. Driessen and C. Hokke and M. Yazdanbakhsh and D. Bosch and A. Schots",
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Westerhof, LB, Wilbers, RHP, van Raaij, DR, Capuder, E, van der Schuren, A, Nguyen, D, Hussaarts, L, Driessen, N, Hokke, C, Yazdanbakhsh, M, Bosch, D & Schots, A 2014, Expression of Schistosome-derived Omega-1 with diantennary glycans carrying Lewis X motifs in Nicotiana benthamiana plants. in Proceedings of The First Conference of the International Society for Plant Molecular Farming. pp. 22-22, The First Conference of the International Society for Plant Molecular Farming, Berlin-Dahlem, Germany, 17/06/14.

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

Proceedings of The First Conference of the International Society for Plant Molecular Farming. 2014. p. 22-22.

Research output: Chapter in Book/Report/Conference proceedingAbstract

TY - CHAP

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

AU - Westerhof, L.B.

AU - Wilbers, R.H.P.

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.

AU - Yazdanbakhsh, M.

AU - Bosch, D.

AU - Schots, A.

PY - 2014

Y1 - 2014

N2 - Plants are an excellent platform for the heterologous production of (glyco)proteins. The surprising tolerance of plants to glyco-engineering allows the production of many different N-glycans. N-glycans play a key role in the interaction between helminth secreted proteins and the host immune system. The exact role of N-glycans on the immunomodulatory properties of helminth secreted glycoproteins remains to be elucidated. Studying the biological function of helminth secreted glycoproteins is challenging, as the purification of a single glycoprotein from helminths or their eggs is relatively inefficient. Plants are an appropriate platform to express these proteins in large quantities with their native N-glycan structures. Omega-1 is a T2 ribonuclease (RNase) secreted by Schistosoma mansoni eggs and is a key factor for the induction of T-helper 2 (Th2) cell differentiation. Induction of Th2 responses by Omega-1 depends on its RNase activity as well as on N-glycan mediated internalisation by antigen presenting cells. Wild type 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 and the mannose receptor. In this study we efficiently produced Schistosome-derived Omega-1 in Nicotiana benthamiana by means of agroinfiltration. The protein 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 - Plants are an excellent platform for the heterologous production of (glyco)proteins. The surprising tolerance of plants to glyco-engineering allows the production of many different N-glycans. N-glycans play a key role in the interaction between helminth secreted proteins and the host immune system. The exact role of N-glycans on the immunomodulatory properties of helminth secreted glycoproteins remains to be elucidated. Studying the biological function of helminth secreted glycoproteins is challenging, as the purification of a single glycoprotein from helminths or their eggs is relatively inefficient. Plants are an appropriate platform to express these proteins in large quantities with their native N-glycan structures. Omega-1 is a T2 ribonuclease (RNase) secreted by Schistosoma mansoni eggs and is a key factor for the induction of T-helper 2 (Th2) cell differentiation. Induction of Th2 responses by Omega-1 depends on its RNase activity as well as on N-glycan mediated internalisation by antigen presenting cells. Wild type 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 and the mannose receptor. In this study we efficiently produced Schistosome-derived Omega-1 in Nicotiana benthamiana by means of agroinfiltration. The protein 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 - 22

EP - 22

BT - Proceedings of The First Conference of the International Society for Plant Molecular Farming

ER -

Westerhof LB, Wilbers RHP, van Raaij DR, Capuder E, van der Schuren A, Nguyen D et al. Expression of Schistosome-derived Omega-1 with diantennary glycans carrying Lewis X motifs in Nicotiana benthamiana plants. In Proceedings of The First Conference of the International Society for Plant Molecular Farming. 2014. p. 22-22