Production and characterization of an orally immunogenic Plasmodium antigen in plants using a virus-based expression system

Diane E. Webster, Lina Wang, Mark Mulcair, Charles Ma, Luca Santi, Hugh Mason, Steve L. Wesselingh, Ross L. Coppel

Research output: Contribution to journalArticle

31 Citations (Scopus)

Abstract

Increasing numbers of plant-made vaccines and pharmaceuticals are entering the late stage of product development and commercialization. Despite the theoretical benefits of such production, expression of parasite antigens in plants, particularly those from Plasmodium, the causative parasites for malaria, have achieved only limited success. We have previously shown that stable transformation of tobacco plants with a plant-codon optimized form of the Plasmodium yoelii merozoite surface protein 4/5 (PyMSP4/5) gene resulted in PyMSP4/5 expression of up to ∼0.25% of total soluble protein. In this report, we describe the rapid expression of PyMSP4/5 in Nicotiana benthamiana leaves using the deconstructed tobacco mosaic virus-based magnICON ® expression system. PyMSP4/5 yields of up to 10% TSP or 1-2 mg/g of fresh weight were consistently achieved. Characterization of the recombinant plant-made PyMSP4/5 indicates that it is structurally similar to PyMSP4/5 expressed by Escherichia coli. It is notable that the plant-made PyMSP4/5 protein retained its immunogenicity following long-term storage at ambient temperature within freeze-dried leaves. With assistance from a mucosal adjuvant the PyMSP4/5-containing leaves induced PyMSP4/5-specific antibodies when delivered orally to naïve mice or mice primed by a DNA vaccine. This study provides evidence that immunogenic Plasmodium antigens can be produced in large quantities in plants using the magnICON® viral vector system.

Original languageEnglish (US)
Pages (from-to)846-855
Number of pages10
JournalPlant Biotechnology Journal
Volume7
Issue number9
DOIs
StatePublished - Dec 2009

Fingerprint

Plant Antigens
Plasmodium yoelii
merozoites
Plasmodium
plant viruses
surface proteins
Viruses
antigens
Tobacco
Parasites
Thrombospondin 1
Tobacco Mosaic Virus
Plasmodium merozoite surface protein 4
merozoite surface protein 5
parasites
leaves
DNA Vaccines
Tobacco mosaic virus
Nicotiana benthamiana
product development

Keywords

  • Immunogenicity
  • Malaria
  • Plant made vaccine
  • Plasmodium
  • PyMSP4/5
  • Viral vector

ASJC Scopus subject areas

  • Plant Science
  • Biotechnology
  • Agronomy and Crop Science

Cite this

Production and characterization of an orally immunogenic Plasmodium antigen in plants using a virus-based expression system. / Webster, Diane E.; Wang, Lina; Mulcair, Mark; Ma, Charles; Santi, Luca; Mason, Hugh; Wesselingh, Steve L.; Coppel, Ross L.

In: Plant Biotechnology Journal, Vol. 7, No. 9, 12.2009, p. 846-855.

Research output: Contribution to journalArticle

Webster, Diane E. ; Wang, Lina ; Mulcair, Mark ; Ma, Charles ; Santi, Luca ; Mason, Hugh ; Wesselingh, Steve L. ; Coppel, Ross L. / Production and characterization of an orally immunogenic Plasmodium antigen in plants using a virus-based expression system. In: Plant Biotechnology Journal. 2009 ; Vol. 7, No. 9. pp. 846-855.
@article{938a0e6d6d6f4b688de34a542eca1231,
title = "Production and characterization of an orally immunogenic Plasmodium antigen in plants using a virus-based expression system",
abstract = "Increasing numbers of plant-made vaccines and pharmaceuticals are entering the late stage of product development and commercialization. Despite the theoretical benefits of such production, expression of parasite antigens in plants, particularly those from Plasmodium, the causative parasites for malaria, have achieved only limited success. We have previously shown that stable transformation of tobacco plants with a plant-codon optimized form of the Plasmodium yoelii merozoite surface protein 4/5 (PyMSP4/5) gene resulted in PyMSP4/5 expression of up to ∼0.25{\%} of total soluble protein. In this report, we describe the rapid expression of PyMSP4/5 in Nicotiana benthamiana leaves using the deconstructed tobacco mosaic virus-based magnICON {\circledR} expression system. PyMSP4/5 yields of up to 10{\%} TSP or 1-2 mg/g of fresh weight were consistently achieved. Characterization of the recombinant plant-made PyMSP4/5 indicates that it is structurally similar to PyMSP4/5 expressed by Escherichia coli. It is notable that the plant-made PyMSP4/5 protein retained its immunogenicity following long-term storage at ambient temperature within freeze-dried leaves. With assistance from a mucosal adjuvant the PyMSP4/5-containing leaves induced PyMSP4/5-specific antibodies when delivered orally to na{\"i}ve mice or mice primed by a DNA vaccine. This study provides evidence that immunogenic Plasmodium antigens can be produced in large quantities in plants using the magnICON{\circledR} viral vector system.",
keywords = "Immunogenicity, Malaria, Plant made vaccine, Plasmodium, PyMSP4/5, Viral vector",
author = "Webster, {Diane E.} and Lina Wang and Mark Mulcair and Charles Ma and Luca Santi and Hugh Mason and Wesselingh, {Steve L.} and Coppel, {Ross L.}",
year = "2009",
month = "12",
doi = "10.1111/j.1467-7652.2009.00447.x",
language = "English (US)",
volume = "7",
pages = "846--855",
journal = "Plant Biotechnology Journal",
issn = "1467-7644",
publisher = "Wiley-Blackwell",
number = "9",

}

TY - JOUR

T1 - Production and characterization of an orally immunogenic Plasmodium antigen in plants using a virus-based expression system

AU - Webster, Diane E.

AU - Wang, Lina

AU - Mulcair, Mark

AU - Ma, Charles

AU - Santi, Luca

AU - Mason, Hugh

AU - Wesselingh, Steve L.

AU - Coppel, Ross L.

PY - 2009/12

Y1 - 2009/12

N2 - Increasing numbers of plant-made vaccines and pharmaceuticals are entering the late stage of product development and commercialization. Despite the theoretical benefits of such production, expression of parasite antigens in plants, particularly those from Plasmodium, the causative parasites for malaria, have achieved only limited success. We have previously shown that stable transformation of tobacco plants with a plant-codon optimized form of the Plasmodium yoelii merozoite surface protein 4/5 (PyMSP4/5) gene resulted in PyMSP4/5 expression of up to ∼0.25% of total soluble protein. In this report, we describe the rapid expression of PyMSP4/5 in Nicotiana benthamiana leaves using the deconstructed tobacco mosaic virus-based magnICON ® expression system. PyMSP4/5 yields of up to 10% TSP or 1-2 mg/g of fresh weight were consistently achieved. Characterization of the recombinant plant-made PyMSP4/5 indicates that it is structurally similar to PyMSP4/5 expressed by Escherichia coli. It is notable that the plant-made PyMSP4/5 protein retained its immunogenicity following long-term storage at ambient temperature within freeze-dried leaves. With assistance from a mucosal adjuvant the PyMSP4/5-containing leaves induced PyMSP4/5-specific antibodies when delivered orally to naïve mice or mice primed by a DNA vaccine. This study provides evidence that immunogenic Plasmodium antigens can be produced in large quantities in plants using the magnICON® viral vector system.

AB - Increasing numbers of plant-made vaccines and pharmaceuticals are entering the late stage of product development and commercialization. Despite the theoretical benefits of such production, expression of parasite antigens in plants, particularly those from Plasmodium, the causative parasites for malaria, have achieved only limited success. We have previously shown that stable transformation of tobacco plants with a plant-codon optimized form of the Plasmodium yoelii merozoite surface protein 4/5 (PyMSP4/5) gene resulted in PyMSP4/5 expression of up to ∼0.25% of total soluble protein. In this report, we describe the rapid expression of PyMSP4/5 in Nicotiana benthamiana leaves using the deconstructed tobacco mosaic virus-based magnICON ® expression system. PyMSP4/5 yields of up to 10% TSP or 1-2 mg/g of fresh weight were consistently achieved. Characterization of the recombinant plant-made PyMSP4/5 indicates that it is structurally similar to PyMSP4/5 expressed by Escherichia coli. It is notable that the plant-made PyMSP4/5 protein retained its immunogenicity following long-term storage at ambient temperature within freeze-dried leaves. With assistance from a mucosal adjuvant the PyMSP4/5-containing leaves induced PyMSP4/5-specific antibodies when delivered orally to naïve mice or mice primed by a DNA vaccine. This study provides evidence that immunogenic Plasmodium antigens can be produced in large quantities in plants using the magnICON® viral vector system.

KW - Immunogenicity

KW - Malaria

KW - Plant made vaccine

KW - Plasmodium

KW - PyMSP4/5

KW - Viral vector

UR - http://www.scopus.com/inward/record.url?scp=70449374925&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=70449374925&partnerID=8YFLogxK

U2 - 10.1111/j.1467-7652.2009.00447.x

DO - 10.1111/j.1467-7652.2009.00447.x

M3 - Article

C2 - 19781007

AN - SCOPUS:70449374925

VL - 7

SP - 846

EP - 855

JO - Plant Biotechnology Journal

JF - Plant Biotechnology Journal

SN - 1467-7644

IS - 9

ER -