title: Production and glyco-engineering of immunomodulatory helminth glycoproteins in plants.
abstract: Helminth parasites control host-immune responses by secreting immunomodulatory glycoproteins. Clinical trials and mouse model studies have demonstrated the potential of helminth-derived glycoproteins for the treatment of immune-related diseases, like allergies and autoimmune diseases. Studies are however hampered by the limited availability of native parasite-derived proteins. Moreover, recombinant protein production systems have thus far been unable to reconstitute helminth-like glycosylation essential for the functionality of some helminth glycoproteins. Here we exploited the flexibility of the N-glycosylation machinery of plants to reconstruct the helminth glycoproteins omega-1 and kappa-5, two major constituents of immunomodulatory Schistosoma mansoni soluble egg antigens. Fine-tuning transient co-expression of specific glycosyltransferases in Nicotiana benthamiana enabled the synthesis of Lewis X (LeX) and LDN/LDN-F glycan motifs as found on natural omega-1 and kappa-5, respectively. In vitro and in vivo evaluation of the introduction of native LeX motifs on plant-produced omega-1 confirmed that LeX on omega-1 contributes to the glycoprotein's Th2-inducing properties. These data indicate that mimicking the complex carbohydrate structures of helminths in plants is a promising strategy to allow targeted evaluation of therapeutic glycoproteins for the treatment of inflammatory disorders. In addition, our results offer perspectives for the development of effective anti-helminthic vaccines by reconstructing native parasite glycoprotein antigens.
authors.affiliation: Laboratory of Nematology, Plant Sciences Group, Wageningen University and Research, Droevendaalsesteeg 1, 6708 PB Wageningen, The Netherlands.
authors.forename: Ruud H P
authors.initials: RH
authors.lastname: Wilbers
authors.affiliation: Laboratory of Nematology, Plant Sciences Group, Wageningen University and Research, Droevendaalsesteeg 1, 6708 PB Wageningen, The Netherlands.
authors.forename: Lotte B
authors.initials: LB
authors.lastname: Westerhof
authors.affiliation: Laboratory of Nematology, Plant Sciences Group, Wageningen University and Research, Droevendaalsesteeg 1, 6708 PB Wageningen, The Netherlands.
authors.forename: Kim
authors.initials: K
authors.lastname: van Noort
authors.affiliation: Department of Parasitology, Leiden University Medical Center, Albinusdreef 2, 2333 ZA Leiden, The Netherlands.
authors.forename: Katja
authors.initials: K
authors.lastname: Obieglo
authors.affiliation: Department of Parasitology, Leiden University Medical Center, Albinusdreef 2, 2333 ZA Leiden, The Netherlands.
authors.forename: Nicole N
authors.initials: NN
authors.lastname: Driessen
authors.affiliation: Department of Parasitology, Leiden University Medical Center, Albinusdreef 2, 2333 ZA Leiden, The Netherlands.
authors.forename: Bart
authors.initials: B
authors.lastname: Everts
authors.affiliation: Department of Experimental Immunology, Academic Medical Center, University of Amsterdam, Meibergdreef 9, 1105 AZ Amsterdam, The Netherlands.
authors.forename: Sonja I
authors.initials: SI
authors.lastname: Gringhuis
authors.affiliation: Research Center Borstel, Priority Area Asthma and Allergy, Experimental Pneumology, Parkallee 22, D-23845, Borstel, Germany.
authors.forename: Gabriele
authors.initials: G
authors.lastname: Schramm
authors.affiliation: Laboratory of Nematology, Plant Sciences Group, Wageningen University and Research, Droevendaalsesteeg 1, 6708 PB Wageningen, The Netherlands.
authors.forename: Aska
authors.initials: A
authors.lastname: Goverse
authors.affiliation: Laboratory of Nematology, Plant Sciences Group, Wageningen University and Research, Droevendaalsesteeg 1, 6708 PB Wageningen, The Netherlands.
authors.forename: Geert
authors.initials: G
authors.lastname: Smant
authors.affiliation: Laboratory of Nematology, Plant Sciences Group, Wageningen University and Research, Droevendaalsesteeg 1, 6708 PB Wageningen, The Netherlands.
authors.forename: Jaap
authors.initials: J
authors.lastname: Bakker
authors.affiliation: Department of Parasitology, Leiden University Medical Center, Albinusdreef 2, 2333 ZA Leiden, The Netherlands.
authors.forename: Hermelijn H
authors.initials: HH
authors.lastname: Smits
authors.affiliation: Department of Parasitology, Leiden University Medical Center, Albinusdreef 2, 2333 ZA Leiden, The Netherlands.
authors.forename: Maria
authors.initials: M
authors.lastname: Yazdanbakhsh
authors.affiliation: Laboratory of Nematology, Plant Sciences Group, Wageningen University and Research, Droevendaalsesteeg 1, 6708 PB Wageningen, The Netherlands.
authors.forename: Arjen
authors.initials: A
authors.lastname: Schots
authors.affiliation: Department of Parasitology, Leiden University Medical Center, Albinusdreef 2, 2333 ZA Leiden, The Netherlands.
authors.forename: Cornelis H
authors.initials: CH
authors.lastname: Hokke
mesh_terms: D000818:Animals; D000909:Antibodies, Helminth / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D000947:Antigens, Helminth / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D004527:Egg Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D015870:Gene Expression / Q000276:immunology; D005818:Genetic Engineering; D006023:Glycoproteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D006031:Glycosylation; D015801:Helminth Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D056747:Immunomodulation / Q000235:genetics / Q000276:immunology; D011994:Recombinant Proteins / Q000276:immunology / Q000378:metabolism; D012550:Schistosoma mansoni / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D014026:Nicotiana / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D014612:Vaccines / Q000276:immunology
chemical_list: D000909:Antibodies, Helminth; D000947:Antigens, Helminth; D004527:Egg Proteins; D006023:Glycoproteins; D015801:Helminth Proteins; D011994:Recombinant Proteins; D014612:Vaccines