Recombinant plant-expressed tumour-associated MUC1 peptide is immunogenic and capable of breaking tolerance in MUC1.Tg mice

Julia Pinkhasov, M. Lucrecia Alvarez, M. Manuela Rigano, Khanrat Piensook, Dalia Larios, Martin Pabst, Josephine Grass, Pinku Mukherjee, Sandra J Gendler, Amanda M. Walmsley, Hugh S. Mason

Research output: Contribution to journalArticle

30 Citations (Scopus)

Abstract

The human epithelial mucin MUC1 is a heavily glycosylated transmembrane protein that is overexpressed and aberrantly glycosylated on over 90% of human breast cancers. The altered glycosylation of MUC1 reveals an immunodominant peptide along its tandem repeat (TR) that has been used as a target for tumour immunotherapy. In this study, we used the MUC1 TR peptide as a test antigen to determine whether a plant-expressed human tumour-associated antigen can be successfully expressed in a plant system and whether it will be able to break self-antigen tolerance in a MUC1-tolerant mouse model. We report the expression of MUC1 TR peptide fused to the mucosal-targeting Escherichia coli enterotoxin B subunit (LTB-MUC1) in a plant host. Utilizing a rapid viral replicon transient expression system, we obtained high yields of LTB-MUC1. Importantly, the LTB-MUC1 fusion protein displayed post-translational modifications that affected its antigenicity. Glycan analysis revealed that LTB-MUC1 was glycosylated and a MUC1-specific monoclonal antibody detected only the glycosylated forms. A thorough saccharide analysis revealed that the glycans are tri-arabinans linked to hydroxyprolines within the MUC1 tandem repeat sequence. We immunized MUC1-tolerant mice (MUC1.Tg) with transiently expressed LTB-MUC1, and observed production of anti-MUC1 serum antibodies, indicating breach of tolerance. The results indicate that a plant-derived human tumour-associated antigen is equivalent to the human antigen in the context of immune recognition.

Original languageEnglish (US)
Pages (from-to)991-1001
Number of pages11
JournalPlant Biotechnology Journal
Volume9
Issue number9
DOIs
StatePublished - Dec 2011

Fingerprint

Plant Tumors
tandem repeat sequences
peptides
antigens
Peptides
neoplasms
mice
Tandem Repeat Sequences
Neoplasm Antigens
Polysaccharides
polysaccharides
arabinans
Antigens
replicon
Replicon
transmembrane proteins
mucins
Enterotoxins
hydroxyproline
immunotherapy

Keywords

  • Cancer vaccine
  • Epithelial mucin 1
  • Heat-labile enterotoxin B subunit
  • Plant expression

ASJC Scopus subject areas

  • Plant Science
  • Biotechnology
  • Agronomy and Crop Science

Cite this

Recombinant plant-expressed tumour-associated MUC1 peptide is immunogenic and capable of breaking tolerance in MUC1.Tg mice. / Pinkhasov, Julia; Alvarez, M. Lucrecia; Rigano, M. Manuela; Piensook, Khanrat; Larios, Dalia; Pabst, Martin; Grass, Josephine; Mukherjee, Pinku; Gendler, Sandra J; Walmsley, Amanda M.; Mason, Hugh S.

In: Plant Biotechnology Journal, Vol. 9, No. 9, 12.2011, p. 991-1001.

Research output: Contribution to journalArticle

Pinkhasov, J, Alvarez, ML, Rigano, MM, Piensook, K, Larios, D, Pabst, M, Grass, J, Mukherjee, P, Gendler, SJ, Walmsley, AM & Mason, HS 2011, 'Recombinant plant-expressed tumour-associated MUC1 peptide is immunogenic and capable of breaking tolerance in MUC1.Tg mice', Plant Biotechnology Journal, vol. 9, no. 9, pp. 991-1001. https://doi.org/10.1111/j.1467-7652.2011.00614.x
Pinkhasov, Julia ; Alvarez, M. Lucrecia ; Rigano, M. Manuela ; Piensook, Khanrat ; Larios, Dalia ; Pabst, Martin ; Grass, Josephine ; Mukherjee, Pinku ; Gendler, Sandra J ; Walmsley, Amanda M. ; Mason, Hugh S. / Recombinant plant-expressed tumour-associated MUC1 peptide is immunogenic and capable of breaking tolerance in MUC1.Tg mice. In: Plant Biotechnology Journal. 2011 ; Vol. 9, No. 9. pp. 991-1001.
@article{d74820a22ea545788cb5952d10545792,
title = "Recombinant plant-expressed tumour-associated MUC1 peptide is immunogenic and capable of breaking tolerance in MUC1.Tg mice",
abstract = "The human epithelial mucin MUC1 is a heavily glycosylated transmembrane protein that is overexpressed and aberrantly glycosylated on over 90{\%} of human breast cancers. The altered glycosylation of MUC1 reveals an immunodominant peptide along its tandem repeat (TR) that has been used as a target for tumour immunotherapy. In this study, we used the MUC1 TR peptide as a test antigen to determine whether a plant-expressed human tumour-associated antigen can be successfully expressed in a plant system and whether it will be able to break self-antigen tolerance in a MUC1-tolerant mouse model. We report the expression of MUC1 TR peptide fused to the mucosal-targeting Escherichia coli enterotoxin B subunit (LTB-MUC1) in a plant host. Utilizing a rapid viral replicon transient expression system, we obtained high yields of LTB-MUC1. Importantly, the LTB-MUC1 fusion protein displayed post-translational modifications that affected its antigenicity. Glycan analysis revealed that LTB-MUC1 was glycosylated and a MUC1-specific monoclonal antibody detected only the glycosylated forms. A thorough saccharide analysis revealed that the glycans are tri-arabinans linked to hydroxyprolines within the MUC1 tandem repeat sequence. We immunized MUC1-tolerant mice (MUC1.Tg) with transiently expressed LTB-MUC1, and observed production of anti-MUC1 serum antibodies, indicating breach of tolerance. The results indicate that a plant-derived human tumour-associated antigen is equivalent to the human antigen in the context of immune recognition.",
keywords = "Cancer vaccine, Epithelial mucin 1, Heat-labile enterotoxin B subunit, Plant expression",
author = "Julia Pinkhasov and Alvarez, {M. Lucrecia} and Rigano, {M. Manuela} and Khanrat Piensook and Dalia Larios and Martin Pabst and Josephine Grass and Pinku Mukherjee and Gendler, {Sandra J} and Walmsley, {Amanda M.} and Mason, {Hugh S.}",
year = "2011",
month = "12",
doi = "10.1111/j.1467-7652.2011.00614.x",
language = "English (US)",
volume = "9",
pages = "991--1001",
journal = "Plant Biotechnology Journal",
issn = "1467-7644",
publisher = "Wiley-Blackwell",
number = "9",

}

TY - JOUR

T1 - Recombinant plant-expressed tumour-associated MUC1 peptide is immunogenic and capable of breaking tolerance in MUC1.Tg mice

AU - Pinkhasov, Julia

AU - Alvarez, M. Lucrecia

AU - Rigano, M. Manuela

AU - Piensook, Khanrat

AU - Larios, Dalia

AU - Pabst, Martin

AU - Grass, Josephine

AU - Mukherjee, Pinku

AU - Gendler, Sandra J

AU - Walmsley, Amanda M.

AU - Mason, Hugh S.

PY - 2011/12

Y1 - 2011/12

N2 - The human epithelial mucin MUC1 is a heavily glycosylated transmembrane protein that is overexpressed and aberrantly glycosylated on over 90% of human breast cancers. The altered glycosylation of MUC1 reveals an immunodominant peptide along its tandem repeat (TR) that has been used as a target for tumour immunotherapy. In this study, we used the MUC1 TR peptide as a test antigen to determine whether a plant-expressed human tumour-associated antigen can be successfully expressed in a plant system and whether it will be able to break self-antigen tolerance in a MUC1-tolerant mouse model. We report the expression of MUC1 TR peptide fused to the mucosal-targeting Escherichia coli enterotoxin B subunit (LTB-MUC1) in a plant host. Utilizing a rapid viral replicon transient expression system, we obtained high yields of LTB-MUC1. Importantly, the LTB-MUC1 fusion protein displayed post-translational modifications that affected its antigenicity. Glycan analysis revealed that LTB-MUC1 was glycosylated and a MUC1-specific monoclonal antibody detected only the glycosylated forms. A thorough saccharide analysis revealed that the glycans are tri-arabinans linked to hydroxyprolines within the MUC1 tandem repeat sequence. We immunized MUC1-tolerant mice (MUC1.Tg) with transiently expressed LTB-MUC1, and observed production of anti-MUC1 serum antibodies, indicating breach of tolerance. The results indicate that a plant-derived human tumour-associated antigen is equivalent to the human antigen in the context of immune recognition.

AB - The human epithelial mucin MUC1 is a heavily glycosylated transmembrane protein that is overexpressed and aberrantly glycosylated on over 90% of human breast cancers. The altered glycosylation of MUC1 reveals an immunodominant peptide along its tandem repeat (TR) that has been used as a target for tumour immunotherapy. In this study, we used the MUC1 TR peptide as a test antigen to determine whether a plant-expressed human tumour-associated antigen can be successfully expressed in a plant system and whether it will be able to break self-antigen tolerance in a MUC1-tolerant mouse model. We report the expression of MUC1 TR peptide fused to the mucosal-targeting Escherichia coli enterotoxin B subunit (LTB-MUC1) in a plant host. Utilizing a rapid viral replicon transient expression system, we obtained high yields of LTB-MUC1. Importantly, the LTB-MUC1 fusion protein displayed post-translational modifications that affected its antigenicity. Glycan analysis revealed that LTB-MUC1 was glycosylated and a MUC1-specific monoclonal antibody detected only the glycosylated forms. A thorough saccharide analysis revealed that the glycans are tri-arabinans linked to hydroxyprolines within the MUC1 tandem repeat sequence. We immunized MUC1-tolerant mice (MUC1.Tg) with transiently expressed LTB-MUC1, and observed production of anti-MUC1 serum antibodies, indicating breach of tolerance. The results indicate that a plant-derived human tumour-associated antigen is equivalent to the human antigen in the context of immune recognition.

KW - Cancer vaccine

KW - Epithelial mucin 1

KW - Heat-labile enterotoxin B subunit

KW - Plant expression

UR - http://www.scopus.com/inward/record.url?scp=80955158806&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=80955158806&partnerID=8YFLogxK

U2 - 10.1111/j.1467-7652.2011.00614.x

DO - 10.1111/j.1467-7652.2011.00614.x

M3 - Article

C2 - 21740504

AN - SCOPUS:80955158806

VL - 9

SP - 991

EP - 1001

JO - Plant Biotechnology Journal

JF - Plant Biotechnology Journal

SN - 1467-7644

IS - 9

ER -