Vaccinia virus recombinants expressing rabiesvirus glycoprotein protect against rabies

Abstract

Six recombinants of New York Board of Health (NYBH) vaccinia virus containing cDNA for Challenge Virus Standard (CVS) rabiesvirus glycoprotein (G) were produced by directing gene insertion into the vaccinia thymidine kinase (TK) locus. To regulate expression of G the promoter P7.5 (functions at early and late times postinfection) from the gene for the vaccinia 7.5 kilodalton (kD) protein was used in two of the recombinants; late promoter P11 of the vaccinia 11 kD protein was used in four recombinants. The six differed in nucleotide sequences flanking the translation start codon; in two constructs the encoded signal peptide of G was fused to several additional amino acids. Cells infected with each recombinant made G that reacted with G-specific antibodies, comigrated with authentic G, and was transported to the plasma membrane. The highest amounts of G were made with fusion or standard versions of G with P11 provided that the mRNA leader sequences were identical to the natural gene. Each recombinant in mice and one in dogs induced rabiesvirus neutralizing antibodies and protection against lethal rabiesvirus challenge.