Salmonella Enteritidis ghost vaccine carrying the hemagglutinin globular head (HA1) domain from H1N1 virus protects against salmonellosis and influenza in chickens

This study evaluated the attenuated Salmonella Enteritidis (SE) ghost strain JOL2114 (Δlon ΔcpxR Δasd), which displays on the bacterial surface the H1N1 hemagglutinin globular head portion (HA1; amino acid residues 63-286) on the bacterial surface for protective efficacy against Salmonella and H1N1 challenge in the chicken model, as the birds are the predominant reservoirs for both diseases. The ghost system enhanced the lysis process by converging two lysis processes found in bacteriophages: bacteriophage PhiX174 lysis gene E and holin-endolysin genes found in bacteriophage λ, complemented with accessory lysis-related proteins Rz/Rz1. The present lysis machinery resulted in complete lysis of host-attenuated SE strains in about 24 hrs of incubation under a non-permissible temperature of 42 °C in the absence of L-arabinose, an antisense inducer that blocks lysis gene expression during the growth phase. SE ghost JOL2114 surface display of HA1 was confirmed by Western blot analysis resulting in an immune-reactive band of 31 kDa in size. Chicken immunization via intramuscular and oral routes yielded both SE and HA1 antigen-specific immune responses. Protective humoral and cell-mediated immune responses were effectively elicited against both Salmonella and influenza challenge. This efficient strategy of ghost generation employs a dual system of phage lysis for biological generation of SE ghosts that preserves the surface antigenic architecture, offering a rapid and effective way to generate vaccines that could be deployed in urgent circumstances to protect against both Salmonella and influenza infection.