SopB effector protein of Salmonella Typhimurium is translocated in mesenteric lymph nodes during murine salmonellosis.
Salmonella Typhimurium harbors two Salmonella pathogenicity islands (SPIs), each encoding a type three secretion system for virulence proteins. Although there is increasing evidence of postinvasion roles for SPI-1, it has been generally accepted that SPI-1 genes are downregulated following the invasion process. Here, we analyzed the expression and translocation of SopB in vitro, in cell culture and in vivo. To this end, a sopB-FLAG-tagged strain of Salmonella Typhimurium was obtained by epitope tagging. Tagged proteins were detected by sodium dodecyl sulfate polyacrylamide gel electrophoresis and immunoblotting with anti-FLAG antibodies. SopB expression was observed in vitro under cultured conditions that mimic the intestinal niche and different intracellular environments. In agreement, bacteria isolated from infected monolayers expressed and translocated SopB for at least 24 h postinoculation. For in vivo experiments, BALB/c mice were inoculated intraperitoneally with the tagged strain of Salmonella Typhimurium. Infecting bacteria and infected cells were recovered from mesenteric lymph nodes. Our results showed that SopB continues to be synthesized in vivo during 5 days after inoculation. Interestingly, translocation of SopB was detected in the cytosol of cells isolated from lymph nodes 1 day after infection. Altogether, these findings indicate that the expression and translocation of SopB during Salmonella infection is not constrained to the initial host-bacteria encounter in the intestinal environment as defined previously.
Departamento de Microbiología, Parasitología e Immunología, Centro de Estudios Farmacológicos y Botánicos CEFyBO-UBA-CONICET, Facultad de Medicina, Universidad de Buenos Aires, Buenos Aires, Argentina. monicagiaco@yahoo.com.ar
Mónica N
MN
Giacomodonato
Sebastián H
SH
Sarnacki
Mariángeles Noto
MN
Llana
María C
MC
Cerquetti
D000818:Animals; D001426:Bacterial Proteins / Q000378:metabolism*; D002460:Cell Line; D003600:Cytosol / Q000737:chemistry; D022781:Hepatocytes / Q000737:chemistry / Q000382:microbiology; D006801:Humans; D008198:Lymph Nodes / Q000382:microbiology*; D008214:Lymphocytes / Q000737:chemistry / Q000382:microbiology; D051379:Mice; D008807:Mice, Inbred BALB C; D012376:Rodent Diseases / Q000382:microbiology; D012481:Salmonella Infections, Animal / Q000382:microbiology*; D012486:Salmonella typhimurium / Q000472:pathogenicity*; D013997:Time Factors; D037521:Virulence Factors / Q000378:metabolism*
D001426:Bacterial Proteins; D037521:Virulence Factors; C489224:SopB protein, Salmonella