Mosaic RNA phage VLPs carrying domain III of the West Nile virus E protein.

Indulis Cielens, Ludmila Jackevica, Arnis Strods, Andris Kazaks, Velta Ose, Janis Bogans, Paul Pumpens, Regina Renhofa

Molecular biotechnology 2014 May

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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.

Citation

Indulis Cielens, Ludmila Jackevica, Arnis Strods, Andris Kazaks, Velta Ose, Janis Bogans, Paul Pumpens, Regina Renhofa. Mosaic RNA phage VLPs carrying domain III of the West Nile virus E protein. Molecular biotechnology. 2014 May;56(5):459-69