title: Mosaic RNA phage VLPs carrying domain III of the West Nile virus E protein.
abstract: The virus-neutralising domain III (DIII) of the West Nile virus glycoprotein E was exposed on the surface of RNA phage AP205 virus-like particles (VLPs) in mosaic form. For this purpose, a 111 amino acid sequence of DIII was added via amber or opal termination codons to the C-terminus of the AP205 coat protein, and mosaic AP205-DIII VLPs were generated by cultivation in amber- or opal-suppressing Escherichia coli strains. After extensive purification to 95 % homogeneity, mosaic AP205-DIII VLPs retained up to 11-16 % monomers carrying DIII domains. The DIII domains appeared on the VLP surface because they were fully accessible to anti-DIII antibodies. Immunisation of BALB/c mice with AP205-DIII VLPs resulted in the induction of specific anti-DIII antibodies, of which the level was comparable to that of the anti-AP205 antibodies generated against the VLP carrier. The AP205-DIII-induced anti-DIII response was represented by a significant fraction of IgG2 isotype antibodies, in contrast to parallel immunisation with the DIII oligopeptide, which failed to induce IgG2 isotype antibodies. Formulation of AP-205-DIII VLPs in alum adjuvant stimulated the level of the anti-DIII response, but did not alter the fraction of IgG2 isotype antibodies. Mosaic AP205-DIII VLPs could be regarded as a promising prototype of a putative West Nile vaccine.
authors.affiliation: Latvian Biomedical Research and Study Centre, Ratsupites Street 1, Riga, 1067, Latvia.
authors.forename: Indulis
authors.initials: I
authors.lastname: Cielens
authors.forename: Ludmila
authors.initials: L
authors.lastname: Jackevica
authors.forename: Arnis
authors.initials: A
authors.lastname: Strods
authors.forename: Andris
authors.initials: A
authors.lastname: Kazaks
authors.forename: Velta
authors.initials: V
authors.lastname: Ose
authors.forename: Janis
authors.initials: J
authors.lastname: Bogans
authors.forename: Paul
authors.initials: P
authors.lastname: Pumpens
authors.forename: Regina
authors.initials: R
authors.lastname: Renhofa
mesh_terms: D000818:Animals; D057134:Antibodies, Neutralizing / Q000235:genetics / Q000276:immunology; D000914:Antibodies, Viral / Q000235:genetics / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D012317:RNA Phages / Q000235:genetics*; D014759:Viral Envelope Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D014902:West Nile virus / Q000235:genetics* / Q000276:immunology / Q000378:metabolism*
chemical_list: D057134:Antibodies, Neutralizing; D000914:Antibodies, Viral; D014759:Viral Envelope Proteins