Construction and immunogenicity of recombinant Mycobacterium bovis BCG expressing GP5 and M protein of porcine reproductive respiratory syndrome virus.
Mycobacterium bovis BCG was used to express a truncated form of GP5 (lacking the first 30 NH(2)-terminal residues) and M protein of porcine reproductive and respiratory syndrome virus (PRRSV). The PRRSV proteins were expressed in BCG under control of the mycobacterial hsp60 gene promoter either in the mycobacterial cytoplasm (BCGGP5cyt and BCGMcyt) or as MT19-fusion proteins on the mycobacterial surface (BCGGP5surf and BCGMsurf). Mice inoculated with BCGGP5surf and BCGMsurf developed antibodies against the viral proteins at 30 days post-inoculation (dpi) as detected by ELISA and Western blot. By 60 dpi, the animals developed titer of neutralizing antibodies of 8. A PRRSV-specific gamma interferon response was also detected in splenocytes of recombinant BCG-inoculated mice at 60 and 90 dpi. These results indicate that BCG was able to express antigens of PRRSV and elicit an immune response against the viral proteins in mice.
Bastos
Reginaldo G
RG
Department of Veterinary and Biomedical Sciences, University of Nebraska at Lincoln, Lincoln, NE 68583-0905, USA.
Dellagostin
Odir A
OA
Barletta
Raúl G
RG
Doster
Allan R
AR
Nelson
Eric
E
Osorio
Fernando A
FA
D000818:Animals; D000914:Antibodies, Viral / Q000276:immunology; D015153:Blotting, Western; D003001:Cloning, Molecular; D004797:Enzyme-Linked Immunosorbent Assay; D015870:Gene Expression; D005822:Genetic Vectors; D051379:Mice; D008807:Mice, Inbred BALB C; D009163:Mycobacterium bovis / Q000235:genetics*; D010455:Peptides / Q000138:chemical synthesis / Q000737:chemistry / Q000276:immunology; D019316:Porcine respiratory and reproductive syndrome virus / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D011994:Recombinant Proteins / Q000276:immunology; D013552:Swine; D014611:Vaccination; D019444:Vaccines, DNA / Q000276:immunology; D014759:Viral Envelope Proteins / Q000235:genetics / Q000276:immunology*; D014763:Viral Matrix Proteins / Q000235:genetics / Q000276:immunology*
D000914:Antibodies, Viral; D010455:Peptides; D011994:Recombinant Proteins; D019444:Vaccines, DNA; D014759:Viral Envelope Proteins; D014763:Viral Matrix Proteins; C114789:glycoprotein 5, PRRSV