Immunization of mice against Salmonella typhimurium using different DNA preparations.
Groups of female BALB/c mice were given primary and booster injections of whole genomic DNA extracted from S. typhimurium, P. aeruginosa, or S. aureus. Other groups of mice were immunized in a similar manner with the 1.57kb fragment of the mouse virulence gene (mviA), pTargeT vector (plasmid DNA)/1.57kb construct, pTargeT vector, or saline. Mice in all groups were challenged intraperitoneally with 100 LD50 of S. typhimurium. The bacterial genomic DNA was extracted using the Pure Gene extraction kit. Specific primers were used to amplify the 1.57kb fragment by PCR. The pTargeT Mammalian Expression Vector System was used to prepare the plasmid/ 1.57kb construct. Bacterial genomic DNA extracted from P. aeruginosa and S. aureus appeared to induce non-specific resistance in mice. Specific, in addition to non-specific resistance appeared to be induced when genomic DNA from S. typhimurium was used. There was a prolongation of survival in the groups of mice that received either the 1.57kb fragment or the pTargeT vector/1.57kb construct and 16.67% and 33.34% respectively, of mice in each group survived at 40 days post challenge. None of the mice in the saline control group survived by day 7 post challenge. It is suggested that the non-specific resistance observed in this study might have been due to the adjuvant effect of the non-methylated CpG and other immunostimulatory motifs in bacterial DNA. Specific resistance obtained when genomic DNA from S. typhimurium was used might have been due to minute antigenic contamination, or virulence factor genes other than the mviA gene, might have been expressed in the host, which induced specific immunity.
Kalfayan
L H
LH
Department of Microbiology and Immunology, Faculty of Medicine, American University of Beirut, Lebanon.
Farhat
D
D
El-Nakat
H
H
Matar
G M
GM
Abdelnoor
A M
AM
D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics; D001428:Bacterial Vaccines / Q000235:genetics / Q000276:immunology*; D004269:DNA, Bacterial / Q000276:immunology*; D004268:DNA-Binding Proteins; D004587:Electrophoresis, Agar Gel; D029968:Escherichia coli Proteins; D005260:Female; D005822:Genetic Vectors; D007114:Immunization; D051379:Mice; D008807:Mice, Inbred BALB C; D016133:Polymerase Chain Reaction; D011550:Pseudomonas aeruginosa / Q000235:genetics / Q000276:immunology; D012481:Salmonella Infections, Animal / Q000517:prevention & control*; D012486:Salmonella typhimurium / Q000235:genetics / Q000276:immunology*; D013211:Staphylococcus aureus / Q000235:genetics / Q000276:immunology; D014157:Transcription Factors; D019444:Vaccines, DNA / Q000235:genetics / Q000276:immunology*
D001426:Bacterial Proteins; D001428:Bacterial Vaccines; D004269:DNA, Bacterial; D004268:DNA-Binding Proteins; D029968:Escherichia coli Proteins; D014157:Transcription Factors; D019444:Vaccines, DNA; C098923:rssB protein, E coli