A comparison of virulent and avirulent strains of Salmonella choleraesuis and their ability to invade Vero cell monolayers.
The mechanisms of invasion used by virulent and avirulent Salmonella choleraesuis were compared using a Vero cell invasion assay. Mouse virulent S. choleraesuis strain 38 and avirulent strain 9 were examined for their ability to invade and survive in Vero cells. The assay was performed by S. choleraesuis infection of the Vero cell monolayer alone and in the presence of various treatments applied to the Vero cell monolayers. Intracellular S. choleraesuis colony forming units were then counted to characterize the mechanism of bacterial uptake. Invasion was not affected by colchicine, but was significantly inhibited in the presence of cytochalasins B and D, chloroquine, and dansylcadaverine. Inhibition by the above substances suggested the importance of microfilaments and of receptor recycling in receptor mediated endocytosis. Both bacterial strains had decreased invasion in the presence of mannose and after enzymic treatment with trypsin. Mannose exposure caused a significant 48% decrease in the uptake of virulent S. choleraesuis 38 and a 28% decrease in avirulent S. choleraesuis 9. Inhibition of endosome acidification did not affect the virulent strain 38 as much as it affected avirulent strain 9. Results from these experiments suggested that Vero cell invasion by S. choleraesuis was due to host uptake by receptor mediated endocytosis, and was mediated in part by mannose-sensitive adhesins. Outer membrane proteins were extracted from the virulent and avirulent strain and compared using SDS-PAGE following surface protein labeling with 125I. Virulent S. choleraesuis 38 had a unique 35 kD protein. The outer membrane proteins of both strains were then examined by radio-immunoprecipitation and western blot using guinea pig polyclonal antisera and the 35 kD protein was again found to be unique to the virulent strain 38. Antisera against the 35 kD protein significantly inhibited invasion of Vero cells by S. choleraesuis strain 38.
Roof
M B
MB
Department of Veterinary Microbiology and Preventive Medicine, Iowa State University, Ames 50010.
Kramer
T T
TT
Roth
J A
JA
D008841:Actin Cytoskeleton / Q000382:microbiology; D000818:Animals; D001422:Bacterial Adhesion; D001425:Bacterial Outer Membrane Proteins / Q000032:analysis; D015153:Blotting, Western; D002738:Chloroquine / Q000494:pharmacology; D002846:Chromatography, Affinity; D003078:Colchicine / Q000494:pharmacology; D003572:Cytochalasins / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D007928:Lethal Dose 50; D008358:Mannose / Q000494:pharmacology; D051379:Mice; D008870:Microtubules / Q000382:microbiology; D012475:Salmonella / Q000187:drug effects / Q000472:pathogenicity* / Q000502:physiology; D014357:Trypsin / Q000494:pharmacology; D014709:Vero Cells; D014774:Virulence
D001425:Bacterial Outer Membrane Proteins; D003572:Cytochalasins; D002738:Chloroquine; D014357:Trypsin; D008358:Mannose; D003078:Colchicine