Analysis of a recombinant dengue-2 virus-dengue-3 virus hybrid envelope protein expressed in a secretary baculovirus system

Helle Bielefeldt-Ohmann, David Beasley, David R. Fitzpatrick, John G. Aaskov

Research output: Contribution to journalArticle

34 Citations (Scopus)

Abstract

In a step towards a tetravalent dengue virus subunit vaccine which is economical to produce, highly immunogenic and stable, a hybrid dengue virus envelope (E) protein molecule has been constructed. It consists of 36 amino acids from the membrane protein, the N-terminal 288 amino acids of the dengue-2 virus E protein plus amino acids 289-424 of the dengue-3 virus E protein. It has been engineered for secretory expression by fusion to a mellitin secretory signal sequence and truncation of the hydrophobic transmembrane segment. Using the baculovirus expression system and serum-free conditions, more than 95% of recombinant dengue-2 virus-dengue-3 virus hybrid E protein (rD2D3E) was secreted into the cell culture supernatant in a stable form with multiple features indicative of preserved conformation. The hybrid molecule reacted with a panel of dengue virus- and flavivirus-specific MAbs which recognize linear or conformational epitopes on dengue virions. Human dengue virus-specific antisera also reacted with the protein. The hybrid rD2D3E protein was able to inhibit the in vitro binding of dengue-2 and dengue-3 viruses to human myelomonocytic cells, suggesting that the receptor-binding epitope(s) was preserved. Adjuvant-free immunization with the hybrid protein induced an antibody response to both dengue-2 and dengue-3 virus in outbred mice, comparable in strength to that of individual rD2E and rD3E proteins. Notably, these antibody responses were primarily of the IgG2a and IgG2b isotype. A strong dengue virus cross-reactive T cell response was also induced in the mice, suggesting that dengue virus hybrid E proteins could form the basis of an efficacious multivalent dengue virus vaccine.

Original languageEnglish (US)
Pages (from-to)2723-2733
Number of pages11
JournalJournal of General Virology
Volume78
Issue number11
StatePublished - Nov 1997
Externally publishedYes

Fingerprint

Viral Envelope Proteins
Dengue Virus
Baculoviridae
Dengue
Proteins
Amino Acids
Antibody Formation
Epitopes
Dengue Vaccines
Melitten
Flavivirus
Subunit Vaccines
Protein Sorting Signals
Virion
Immune Sera
Immunization
Membrane Proteins
Cell Culture Techniques

ASJC Scopus subject areas

  • Virology
  • Immunology

Cite this

Analysis of a recombinant dengue-2 virus-dengue-3 virus hybrid envelope protein expressed in a secretary baculovirus system. / Bielefeldt-Ohmann, Helle; Beasley, David; Fitzpatrick, David R.; Aaskov, John G.

In: Journal of General Virology, Vol. 78, No. 11, 11.1997, p. 2723-2733.

Research output: Contribution to journalArticle

Bielefeldt-Ohmann, Helle ; Beasley, David ; Fitzpatrick, David R. ; Aaskov, John G. / Analysis of a recombinant dengue-2 virus-dengue-3 virus hybrid envelope protein expressed in a secretary baculovirus system. In: Journal of General Virology. 1997 ; Vol. 78, No. 11. pp. 2723-2733.
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