Exploiting 2A peptides to elicit potent neutralizing antibodies by a multi-subunit herpesvirus glycoprotein complex

Felix Wussow, Flavia Chiuppesi, Zhuo Meng, Joy Martinez, Jenny Nguyen, Peter A Barry, Don J. Diamond

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

Neutralizing antibodies (NAb) interfering with glycoprotein complex-mediated virus entry into host cells are thought to contribute to the protection against herpesvirus infection. However, using herpesvirus glycoprotein complexes as vaccine antigens can be complicated by the necessity of expressing multiple subunits simultaneously to allow efficient complex assembly and formation of conformational NAb epitopes. By using a novel bacterial artificial chromosome (BAC) clone of the clinically deployable Modified Vaccinia Ankara (MVA) vector and exploiting ribosomal skipping mediated by 2A peptides, MVA vectors were generated that expressed self-processing subunits of the human cytomegalovirus (HCMV) pentamer complex (PC) composed of gH, gL, UL128, UL130, and UL131A. These MVA vectors expressed 2A-linked HCMV PC subunits that were efficiently cleaved and transported to the cell surface as protein complexes forming conformational neutralizing epitopes. In addition, vaccination of mice by only two immunizations with these MVA vectors resulted in potent HCMV NAb responses that remained stable over a period of at least six months. This method of eliciting NAb by 2A-linked, self-processing HCMV PC subunits could contribute to develop a HCMV vaccine candidate and may serve as a template to facilitate the development of subunit vaccine strategies against other herpesviruses.

Original languageEnglish (US)
Pages (from-to)30-37
Number of pages8
JournalJournal of Virological Methods
Volume251
DOIs
StatePublished - Jan 1 2018

Fingerprint

Herpesviridae
Vaccinia
Neutralizing Antibodies
Glycoproteins
Cytomegalovirus
Peptides
Epitopes
Cytomegalovirus Vaccines
Bacterial Artificial Chromosomes
Herpesviridae Infections
Virus Internalization
Subunit Vaccines
Antibody Formation
Immunization
Membrane Proteins
Vaccination
Vaccines
Clone Cells
Antigens

Keywords

  • 2A peptide
  • Artificial chromosome
  • Glycoprotein
  • Herpesvirus
  • Neutralizing antibody
  • Vaccine

ASJC Scopus subject areas

  • Virology

Cite this

Exploiting 2A peptides to elicit potent neutralizing antibodies by a multi-subunit herpesvirus glycoprotein complex. / Wussow, Felix; Chiuppesi, Flavia; Meng, Zhuo; Martinez, Joy; Nguyen, Jenny; Barry, Peter A; Diamond, Don J.

In: Journal of Virological Methods, Vol. 251, 01.01.2018, p. 30-37.

Research output: Contribution to journalArticle

Wussow, Felix ; Chiuppesi, Flavia ; Meng, Zhuo ; Martinez, Joy ; Nguyen, Jenny ; Barry, Peter A ; Diamond, Don J. / Exploiting 2A peptides to elicit potent neutralizing antibodies by a multi-subunit herpesvirus glycoprotein complex. In: Journal of Virological Methods. 2018 ; Vol. 251. pp. 30-37.
@article{32c1253729a740cb90e928b1cdf35b37,
title = "Exploiting 2A peptides to elicit potent neutralizing antibodies by a multi-subunit herpesvirus glycoprotein complex",
abstract = "Neutralizing antibodies (NAb) interfering with glycoprotein complex-mediated virus entry into host cells are thought to contribute to the protection against herpesvirus infection. However, using herpesvirus glycoprotein complexes as vaccine antigens can be complicated by the necessity of expressing multiple subunits simultaneously to allow efficient complex assembly and formation of conformational NAb epitopes. By using a novel bacterial artificial chromosome (BAC) clone of the clinically deployable Modified Vaccinia Ankara (MVA) vector and exploiting ribosomal skipping mediated by 2A peptides, MVA vectors were generated that expressed self-processing subunits of the human cytomegalovirus (HCMV) pentamer complex (PC) composed of gH, gL, UL128, UL130, and UL131A. These MVA vectors expressed 2A-linked HCMV PC subunits that were efficiently cleaved and transported to the cell surface as protein complexes forming conformational neutralizing epitopes. In addition, vaccination of mice by only two immunizations with these MVA vectors resulted in potent HCMV NAb responses that remained stable over a period of at least six months. This method of eliciting NAb by 2A-linked, self-processing HCMV PC subunits could contribute to develop a HCMV vaccine candidate and may serve as a template to facilitate the development of subunit vaccine strategies against other herpesviruses.",
keywords = "2A peptide, Artificial chromosome, Glycoprotein, Herpesvirus, Neutralizing antibody, Vaccine",
author = "Felix Wussow and Flavia Chiuppesi and Zhuo Meng and Joy Martinez and Jenny Nguyen and Barry, {Peter A} and Diamond, {Don J.}",
year = "2018",
month = "1",
day = "1",
doi = "10.1016/j.jviromet.2017.10.006",
language = "English (US)",
volume = "251",
pages = "30--37",
journal = "Journal of Virological Methods",
issn = "0166-0934",
publisher = "Elsevier",

}

TY - JOUR

T1 - Exploiting 2A peptides to elicit potent neutralizing antibodies by a multi-subunit herpesvirus glycoprotein complex

AU - Wussow, Felix

AU - Chiuppesi, Flavia

AU - Meng, Zhuo

AU - Martinez, Joy

AU - Nguyen, Jenny

AU - Barry, Peter A

AU - Diamond, Don J.

PY - 2018/1/1

Y1 - 2018/1/1

N2 - Neutralizing antibodies (NAb) interfering with glycoprotein complex-mediated virus entry into host cells are thought to contribute to the protection against herpesvirus infection. However, using herpesvirus glycoprotein complexes as vaccine antigens can be complicated by the necessity of expressing multiple subunits simultaneously to allow efficient complex assembly and formation of conformational NAb epitopes. By using a novel bacterial artificial chromosome (BAC) clone of the clinically deployable Modified Vaccinia Ankara (MVA) vector and exploiting ribosomal skipping mediated by 2A peptides, MVA vectors were generated that expressed self-processing subunits of the human cytomegalovirus (HCMV) pentamer complex (PC) composed of gH, gL, UL128, UL130, and UL131A. These MVA vectors expressed 2A-linked HCMV PC subunits that were efficiently cleaved and transported to the cell surface as protein complexes forming conformational neutralizing epitopes. In addition, vaccination of mice by only two immunizations with these MVA vectors resulted in potent HCMV NAb responses that remained stable over a period of at least six months. This method of eliciting NAb by 2A-linked, self-processing HCMV PC subunits could contribute to develop a HCMV vaccine candidate and may serve as a template to facilitate the development of subunit vaccine strategies against other herpesviruses.

AB - Neutralizing antibodies (NAb) interfering with glycoprotein complex-mediated virus entry into host cells are thought to contribute to the protection against herpesvirus infection. However, using herpesvirus glycoprotein complexes as vaccine antigens can be complicated by the necessity of expressing multiple subunits simultaneously to allow efficient complex assembly and formation of conformational NAb epitopes. By using a novel bacterial artificial chromosome (BAC) clone of the clinically deployable Modified Vaccinia Ankara (MVA) vector and exploiting ribosomal skipping mediated by 2A peptides, MVA vectors were generated that expressed self-processing subunits of the human cytomegalovirus (HCMV) pentamer complex (PC) composed of gH, gL, UL128, UL130, and UL131A. These MVA vectors expressed 2A-linked HCMV PC subunits that were efficiently cleaved and transported to the cell surface as protein complexes forming conformational neutralizing epitopes. In addition, vaccination of mice by only two immunizations with these MVA vectors resulted in potent HCMV NAb responses that remained stable over a period of at least six months. This method of eliciting NAb by 2A-linked, self-processing HCMV PC subunits could contribute to develop a HCMV vaccine candidate and may serve as a template to facilitate the development of subunit vaccine strategies against other herpesviruses.

KW - 2A peptide

KW - Artificial chromosome

KW - Glycoprotein

KW - Herpesvirus

KW - Neutralizing antibody

KW - Vaccine

UR - http://www.scopus.com/inward/record.url?scp=85030870274&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85030870274&partnerID=8YFLogxK

U2 - 10.1016/j.jviromet.2017.10.006

DO - 10.1016/j.jviromet.2017.10.006

M3 - Article

C2 - 28989096

AN - SCOPUS:85030870274

VL - 251

SP - 30

EP - 37

JO - Journal of Virological Methods

JF - Journal of Virological Methods

SN - 0166-0934

ER -