Bacteriophage T4 nanoparticle capsid surface SOC and HOC bipartite display with enhanced classical swine fever virus immunogenicity: a powerful immunological approach

Abstract

The phage T4 HOC, SOC bipartite display system is attractive for the expression of cDNA and display of peptides or proteins at high copy numbers on the phage capsid surface. Until recently, using T4 phage vector to display large foreign molecular immunogens resulted only from either an SOC or HOC single site. In this report, the main advantages of the phage T4 system over other display technologies are substantiated by using the phage T4 SOC, HOC dual site display vector T4-Zh(-) to express: (1) on the SOC site, the classical swine fever virus (CSFV) major antigenic determinant cluster mE2 (123 amino acid, aa) through gene fusion to the SOC gene C-terminus of T4 genome, and (2) on the HOC site, full-length CSFV primary antigen E2 (371 aa) through a co-transformed plasmid, hence leading to a simultaneous display of both proteins on the T4 capsid surface. The immunogenicities of these constructs were measured by ID-ELISA, dot-ELISA, Western blotting, and immunogenic response in mice including humoral and cellular immunity tests. The immunological efficiencies both in vitro and in mice of phage T4 with both single site and dual site displays, as well as conventional Escherichia coli plasmid expression, were evaluated. The animal immune response data showed that the antibody titers elicited by the T4 phage-CSFV recombinants were significantly higher than those obtained by E. coli plasmid expression, and the unpurified double site display T4 phage particles were around two times higher than either single site display or plasmid expression while being at lower phage concentrations than the single site phages. The immunogens were effective in the absence of eukaryotic protein modifications. Therefore, the phage T4 dual site display emerges as a powerful method with an enhanced immune response in animals for research and development of immunological products.