Influence of porcine reproductive and respiratory syndrome virus GP5 glycoprotein N-linked glycans on immune responses in mice

Porcine reproductive and respiratory syndrome virus (PRRSV) is one of the most economically significant viral diseases in the swine industry. Infection with PRRSV following vaccination is common, since protection is incomplete. Persistent infection may be one of the biggest obstacles to control of the disease. "Glycan shielding" was postulated to be a primary mechanism to explain evasion from neutralizing immune response, ensuring in vivo persistence of virus, such as HIV, SIV, and HBV. The objective of this study was to construct recombinant adenoviruses expressing single or multiple N-linked glycosylation site (NGS) mutant GP5 of PRRSV, and evaluate the expression in cell culture, and potential to induce immune responses in BALB/c mice. Six recombinant adenoviruses were constructed each expressing wild-type GP5 and 1-4 NGS mutants: N44S, N44/51S, N30/44/51S, N30/33/44/51S and N30/33S. Inoculation of BALB/c mice with all five recombinants expressing NGS mutant GP5 resulted in a significant neutralizing antibody responses which were significantly higher than that of recombinant adenovirus expressing wild-type GP5. But there were no significant difference in lymphocyte proliferation responses induced by wild type and NGS mutant GP5. It indicated that glycosylations of GP5 at residues N30, N33, N44 and N51 are critical for induction of neutralizing antibodies. These NGS mutant PRRSV GP5 will be useful to characterize the effects of glycosylation on immunogenicity in the natural host, and may lead to a new approach for the generation of PRRSV vaccines.