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

J Virol Methods. 2018 Jan:251:30-37. doi: 10.1016/j.jviromet.2017.10.006. Epub 2017 Oct 6.

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.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Antibodies, Neutralizing / blood*
  • Antibodies, Viral / blood*
  • Antigens, Viral / genetics
  • Antigens, Viral / immunology*
  • Cytomegalovirus / genetics
  • Cytomegalovirus / immunology*
  • Cytomegalovirus Vaccines / administration & dosage
  • Cytomegalovirus Vaccines / genetics
  • Cytomegalovirus Vaccines / immunology*
  • Genetic Vectors
  • Glycoproteins / genetics
  • Glycoproteins / immunology*
  • Immunization Schedule
  • Mice
  • Vaccines, Synthetic / administration & dosage
  • Vaccines, Synthetic / genetics
  • Vaccines, Synthetic / immunology
  • Vaccinia virus / genetics
  • Viral Structural Proteins / genetics
  • Viral Structural Proteins / immunology*

Substances

  • Antibodies, Neutralizing
  • Antibodies, Viral
  • Antigens, Viral
  • Cytomegalovirus Vaccines
  • Glycoproteins
  • Vaccines, Synthetic
  • Viral Structural Proteins