Cytolethal distending toxin B (cdtB) is a conserved virulence factor in Salmonella enterica serovar Typhi. Here we report the presence and functionality of cdtB in some nontyphoidal Salmonella (NTS) serovars, including Salmonella Javiana (cdtB+wt S. Javiana), isolated from imported food. To understand the role of cdtB in NTS serovars, a deletion mutant (cdtB−ΔS. Javiana) was constructed. Macrophages were infected with cdtB+wt S. Javiana (wild type), cdtB−Δ S. Javiana (mutant), and cdtB-negative NTS serovar (S. Typhimurium). Cytotoxic activity and transcription level of genes involved in cell death (apoptosis, autophagy, and necrosis) were assessed in infected macrophages. The cdtB+wt S. Javiana caused cellular distension as well as high degree of vacuolization and presence of the autophagosome marker LC3 in infected macrophages as compared with cdtB−ΔS. Javiana. The mRNA expression of genes involved in the induction of autophagy in response to toxin (Esr1 and Pik3C3) and coregulators of autophagy and apoptosis (Bax and Cyld) were significantly upregulated in cdtB+wt S. Javiana-infected macrophages. As autophagy destroys internalized pathogens in addition to the infected cell, it may reduce the spread of infection.
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References
1.
AkifusaS., HeywoodW., NairS.P., StenbeckG., and HendersonB. (2005). Mechanism of internalization of the cytolethal distending toxin of Actinobacillus actinomycetemcomitans. Microbiology, 151(Pt 5), 1395–1402.
ArthurC.R., GuptonJ.T., KelloggG.E., YeudallW.A., CabotM.C., NewshamI.F., and GewirtzD.A. (2007). Autophagic cell death, polyploidy and senescence induced in breast tumor cells by the substituted pyrrole JG-03-14, a novel microtubule poison. Biochem Pharmacol, 74, 981–991.
4.
BiswasD., FernandoU., ReimanC., WillsonP., PotterA., and AllanB. (2006). Effect of cytolethal distending toxin of Campylobacter jejuni on adhesion and internalization in cultured cells and in colonization of the chicken gut. Avian Dis, 50, 586–593.
5.
BonapaceL., BornhauserB.C., SchmitzM., CarioG., ZieglerU., NiggliF.K., SchaferB.W., SchrappeM., StanullaM., and BourquinJ.P. (2010). Induction of autophagy-dependent necroptosis is required for childhood acute lymphoblastic leukemia cells to overcome glucocorticoid resistance. J Clin Invest, 120, 1310–1323.
6.
Centers for Disease Control and Prevention (2012) Foodborne Diseases Active Surveillance Network (FoodNet); FoodNet Surveillance Report for 2011 (Final Report), Atlanta, Georgia: U.S. Department of Health and Human Services, CDC.
7.
ChungP.Y., and Van HulW. (2012). Paget's disease of bone: evidence for complex pathogenetic interactions. Semin Arthritis Rheum, 41, 619–641.
8.
ClarksonL.S., Tobin-D'AngeloM., ShulerC., HannaS., BensonJ., and VoetschA.C. (2010). Sporadic Salmonella enterica serotype Javiana infections in Georgia and Tennessee: a hypothesis-generating study. Epidemiol Infect, 138, 340–346.
9.
CoburnB., GrasslG.A., and FinlayB.B. (2007). Salmonella, the host and disease: a brief review. Immunol Cell Biol, 85, 112–118.
DatsenkoK.A., and WannerB.L. (2000). One-step inactivation of chromosomal genes in Escherichia coli K-12 using PCR products. Proc Natl Acad Sci U S A, 97, 6640–6645.
12.
den BakkerH.C., Moreno SwittA.I., GovoniG., CummingsC.A., RanieriM.L., DegoricijaL., HoelzerK., Rodriguez-RiveraL.D., BrownS., BolchacovaE., FurtadoM.R., and WiedmannM. (2011). Genome sequencing reveals diversification of virulence factor content and possible host adaptation in distinct subpopulations of Salmonella enterica. BMC Genomics, 12, 425.
13.
EshraghiA., Maldonado-ArochoF.J., GargiA., CardwellM.M., ProutyM.G., BlankeS.R., and BradleyK.A. (2010). Cytolethal distending toxin family members are differentially affected by alterations in host glycans and membrane cholesterol. J Biol Chem, 285, 18199–18207.
14.
GokulanK., KhareS., RooneyA.W., HanJ., LynneA.M., and FoleyS.L. (2013). Impact of plasmids, including those encodingVirB4/D4 type IV secretion systems, on Salmonella enterica serovar Heidelberg virulence in macrophages and epithelial cells. PLoS One, 8, e77866.
15.
GutierrezM.G., SakaH.A., ChinenI., ZoppinoF.C., YoshimoriT., BoccoJ.L., and ColomboM.I. (2007). Protective role of autophagy against Vibrio cholerae cytolysin, a pore-forming toxin from V. cholerae. Proc Natl Acad Sci U S A, 104, 1829–1834.
16.
HaghjooE., and GalanJ.E. (2004). Salmonella typhi encodes a functional cytolethal distending toxin that is delivered into host cells by a bacterial-internalization pathway. Proc Natl Acad Sci U S A, 101, 4614–4619.
17.
HanJ., GokulanK., BarnetteD., KhareS., RooneyA.W., DeckJ., NayakR., StefanovaR., HartM.E., and FoleyS.L. (2013). Evaluation of virulence and antimicrobial resistance in Salmonella enterica serovar Enteritidis isolates from humans and chicken- and egg-associated sources. Foodborne Pathog Dis, 10, 1008–1015.
HuX., NesicD., and StebbinsC.E. (2006). Comparative structure-function analysis of cytolethal distending toxins. Proteins, 62, 421–434.
20.
HuettA., NgA., CaoZ., KuballaP., KomatsuM., DalyM.J., PodolskyD.K., and XavierR.J. (2009). A novel hybrid yeast-human network analysis reveals an essential role for FNBP1L in antibacterial autophagy. J Immunol, 182, 4917–4930.
21.
JaberN., DouZ., LinR.Z., ZhangJ., and ZongW.X. (2012). Mammalian PIK3C3/VPS34: the key to autophagic processing in liver and heart. Autophagy, 8, 707–708.
22.
JacksonB.R., GriffinP.M., ColeD., WalshK.A., and ChaiS.J. (2013). Outbreak-associated Salmonella enterica serotypes and food Commodities, United States, 1998–2008. Emerg Infect Dis, 19, 1239–1244.
23.
JeffriesC.D., HoltmanD.F., and GuseD.G. (1957). Rapid method for determining the activity of microorganisms on nucleic acids. J Bacteriol, 73, 590–591.
24.
JinadasaR.N., BloomS.E., WeissR.S., and DuhamelG.E. (2011). Cytolethal distending toxin: a conserved bacterial genotoxin that blocks cell cycle progression, leading to apoptosis of a broad range of mammalian cell lineages. Microbiology, 157(Pt 7), 1851–1875.
25.
JohnsonL.R., GouldL.H., DunnJ.R., BerkelmanR., and MahonB.E. (2011). Salmonella infections associated with international travel: a Foodborne Diseases Active Surveillance Network (FoodNet) study. Foodborne Pathog Dis, 8, 1031–1037.
26.
JuhasovaB., Bhatia-KissovaI., PolcicovaK., MentelM., ForteM., and PolcicP. (2011). Reconstitution of interactions of Murine gammaherpesvirus 68 M11 with Bcl-2 family proteins in yeast. Biochem Biophys Res Commun, 407, 783–787.
27.
KalischukL.D., InglisG.D., and BuretA.G. (2007). Strain-dependent induction of epithelial cell oncosis by Campylobacter jejuni is correlated with invasion ability and is independent of cytolethal distending toxin. Microbiology, 153(Pt 9), 2952–2963.
28.
KhareS., FichtT.A., SantosR.L., RomanoJ., FichtA.R., ZhangS., GrantI.R., LibalM., HunterD., and AdamsL.G. (2004). Rapid and sensitive detection of Mycobacterium avium subsp. paratuberculosis in bovine milk and feces by a combination of immunomagnetic bead separation-conventional PCR and real-time PCR. J Clin Microbiol, 42, 1075–1081.
29.
KhareS., LawhonS.D., DrakeK.L., NunesJ.E., FigueiredoJ.F., RossettiC.A., GullT., EvertsR.E., LewinH.A., GalindoC.L., GarnerH.R., and AdamsL.G. (2012). Systems biology analysis of gene expression during in vivo mycobacterium avium paratuberculosis enteric colonization reveals role for immune tolerance. PLoS One, 7, e42127.
30.
Lara-TejeroM., and GalanJ.E. (2000). A bacterial toxin that controls cell cycle progression as a deoxyribonuclease I-like protein. Science, 290, 354–357.
31.
Lara-TejeroM., and GalanJ.E. (2002). Cytolethal distending toxin: limited damage as a strategy to modulate cellular functions. Trends Microbiol, 10, 147–152.
32.
LinY.L., LinS.R., WuT.T., and ChangL.S. (2004). Evidence showing an intermolecular interaction between KChIP proteins and Taiwan cobra cardiotoxins. Biochem Biophys Res Commun, 319, 720–724.
33.
LoriaR.M., and GrafM.R. (2012). 17alpha-androstenediol-mediated oncophagy of tumor cells by different mechanisms is determined by the target tumor. Ann N Y Acad Sci, 1262, 127–133.
34.
MaranC., TassoneE., MasolaV., and OnistoM. (2009). The story of SPATA2 (spermatogenesis-associated protein 2): from Sertoli cells to pancreatic beta-cells. Curr Genomics, 10, 361–363.
35.
McSweeneyL.A., and DreyfusL.A. (2004). Nuclear localization of the Escherichia coli cytolethal distending toxin CdtB subunit. Cell Microbiol, 6, 447–458.
36.
MezalE.H., StefanovaR., and KhanA.A. (2013). Isolation and molecular characterization of Salmonella enterica serovar Javiana from food, environmental and clinical samples. Int J Food Microbiol, 164, 113–118.
NicolaA.M., AlbuquerqueP., MartinezL.R., Dal-RossoR.A., SaylorC., De JesusM., NosanchukJ.D., and CasadevallA. (2012). Macrophage autophagy in immunity to Cryptococcus neoformans and Candida albicans. Infect Immun, 80, 3065–3076.
39.
PeresS.Y., MarchesO., DaigleF., NougayredeJ.P., HeraultF., TascaC., De RyckeJ., and OswaldE. (1997). A new cytolethal distending toxin (CDT) from Escherichia coli producing CNF2 blocks HeLa cell division in G2/M phase. Mol Microbiol, 24, 1095–1107.
40.
RaffatelluM., WilsonR.P., ChessaD., Andrews-PolymenisH., TranQ.T., LawhonS., KhareS., AdamsL.G., and BaumlerA.J. (2005). SipA, SopA, SopB, SopD, and SopE2 contribute to Salmonella enterica serotype typhimurium invasion of epithelial cells. Infect Immun, 73, 146–154.
41.
RikiishiH. (2012). Novel insights into the interplay between apoptosis and autophagy. Int J Cell Biol, 2012, 317645.
42.
RubyT., McLaughlinL., GopinathS., and MonackD. (2012). Salmonella's long-term relationship with its host. FEMS Microbiol Rev, 36, 600–615.
43.
SkybergJ.A., LogueC.M., and NolanL.K. (2006). Virulence genotyping of Salmonella spp. with multiplex PCR. Avian Dis, 50, 77–81.
44.
SmithJ.L., and BaylesD.O. (2006). The contribution of cytolethal distending toxin to bacterial pathogenesis. Crit Rev Microbiol, 32, 227–248.
45.
SongJ., GaoX., and GalanJ.E. (2013). Structure and function of the Salmonella Typhi chimaeric A(2)B(5) typhoid toxin. Nature, 499, 350–354.
46.
SouvannavongV., SaidjiN., and ChabyR. (2007). Lipopolysaccharide from Salmonella enterica activates NF-kappaB through both classical and alternative pathways in primary B Lymphocytes. Infect Immun, 75, 4998–5003.
47.
SpanoS., UgaldeJ.E., and GalanJ.E. (2008). Delivery of a Salmonella Typhi exotoxin from a host intracellular compartment. Cell Host Microbe, 3, 30–38.
48.
SrikantiahP., LayJ.C., HandS., CrumpJ.A., CampbellJ., Van DuyneM.S., BishopR., MiddendorR., CurrierM., MeadP.S., and MolbakK. (2004). Salmonella enterica serotype Javiana infections associated with amphibian contact, Mississippi, 2001. Epidemiol Infect, 132, 273–281.
49.
SuezJ., PorwollikS., DaganA., MarzelA., SchorrY.I., DesaiP.T., AgmonV., McClellandM., RahavG., and Gal-MorO. (2013). Virulence gene profiling and pathogenicity characterization of non-typhoidal Salmonella accounted for invasive disease in humans. PLoS One, 8, e58449.
50.
TsagaratouA., KontoyiannisD.L., and MosialosG. (2011). Truncation of the deubiquitinating domain of CYLD in myelomonocytic cells attenuates inflammatory responses. PLoS One 6,e16397.
51.
van der VeldenA.W., LindgrenS.W., WorleyM.J., and HeffronF. (2000). Salmonella pathogenicity island 1-independent induction of apoptosis in infected macrophages by Salmonella enterica serotype typhimurium. Infect Immun, 68, 5702–5709.
52.
WangW.Y., LimJ.H., and LiJ.D. (2012). Synergistic and feedback signaling mechanisms in the regulation of inflammation in respiratory infections. Cell Mol Immunol, 9, 131–135.
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