Abstract
Measuring bacterial adherence and invasion of cells in vitro has enabled researchers to dissect the interactions of Campylobacter jejuni with eukaryotic cells. Numerous C. jejuni virulence determinants and host cell factors that contribute to the process of adherence, invasion, and immune modulation have been identified utilizing in vitro adherence and invasion assays. In this chapter, we describe the evaluation of C. jejuni adherence to and invasion of HeLa cells using the gentamicin-protection assay.
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References
Allos BM (2001) Campylobacter jejuni infections: update on emerging issues and trends. Clin Infect Dis 32(8):1201–1206
Jin S, Joe A, Lynett J et al (2001) JlpA, a novel surface-exposed lipoprotein specific to Campylobacter jejuni, mediates adherence to host epithelial cells. Mol Microbiol 39(5):1225–1236
Pei Z, Burucoa C, Grignon B et al (1998) Mutation in the peb1A locus of Campylobacter jejuni reduces interactions with epithelial cells and intestinal colonization of mice. Infect Immun 66(3):938–943
Flanagan RC, Neal-McKinney JM, Dhillon AS et al (2009) Examination of Campylobacter jejuni putative adhesins leads to the identification of a new protein, designated FlpA, required for chicken colonization. Infect Immun 77(6):2399–2407. doi:10.1128/IAI.01266-08
Moser I, Schroeder W, Salnikow J (1997) Campylobacter jejuni major outer membrane protein and a 59-kDa protein are involved in binding to fibronectin and INT 407 cell membranes. FEMS Microbiol Lett 157(2):233–238
Konkel ME, Cieplak W Jr (1992) Altered synthetic response of Campylobacter jejuni to cocultivation with human epithelial cells is associated with enhanced internalization. Infect Immun 60(11):4945–4949
Konkel ME, Christensen JE, Dhillon AS et al (2007) Campylobacter jejuni strains compete for colonization in broiler chicks. Appl Environ Microbiol 73(7):2297–2305. doi:10.1128/AEM.02193-06
Konkel ME, Kim BJ, Rivera-Amill V et al (1999) Bacterial secreted proteins are required for the internaliztion of Campylobacter jejuni into cultured mammalian cells. Mol Microbiol 32(4):691–701
Hornef MW, Wick MJ, Rhen M et al (2002) Bacterial strategies for overcoming host innate and adaptive immune responses. Nat Immunol 3(11):1033–1040. doi:10.1038/ni1102-1033
Neal-McKinney JM, Konkel ME (2012) The Campylobacter jejuni CiaC virulence protein is secreted from the flagellum and delivered to the cytosol of host cells. Front Cell Infect Microbiol 2:31. doi:10.3389/fcimb.2012.00031
Vaudaux P, Waldvogel FA (1979) Gentamicin antibacterial activity in the presence of human polymorphonuclear leukocytes. Antimicrob Agents Chemother 16(6):743–749
Weingarten RA, Grimes JL, Olson JW (2008) Role of Campylobacter jejuni respiratory oxidases and reductases in host colonization. Appl Environ Microbiol 74(5):1367–1375. doi:10.1128/AEM.02261-07
Konkel ME, Larson CL, Flanagan RC (2010) Campylobacter jejuni FlpA binds fibronectin and is required for maximal host cell adherence. J Bacteriol 192(1):68–76. doi:10.1128/JB.00969-09
Christensen JE, Pacheco SA, Konkel ME (2009) Identification of a Campylobacter jejuni-secreted protein required for maximal invasion of host cells. Mol Microbiol 73(4):650–662. doi:10.1111/j.1365-2958.2009.06797.x
Samuelson DR, Eucker TP, Bell JA et al (2013) The Campylobacter jejuni CiaD effector protein activates MAP kinase signaling pathways and is required for the development of disease. Cell Commun Signal 11:79. doi:10.1186/1478-811X-11-79
Buelow DR, Christensen JE, Neal-McKinney JM et al (2011) Campylobacter jejuni survival within human epithelial cells is enhanced by the secreted protein CiaI. Mol Microbiol 80(5):1296–1312. doi:10.1111/j.1365-2958.2011.07645.x
Neal-McKinney JM, Christensen JE, Konkel ME (2010) Amino-terminal residues dictate the export efficiency of the Campylobacter jejuni filament proteins via the flagellum. Mol Microbiol 76(4):918–931. doi:10.1111/j.1365-2958.2010.07144.x
Konkel ME, Garvis SG, Tipton SL et al (1997) Identification and molecular cloning of a gene encoding a fibronectin-binding protein (CadF) from Campylobacter jejuni. Mol Microbiol 24(5):953–963
Hu L, Kopecko DJ (1999) Campylobacter jejuni 81-176 associates with microtubules and dynein during invasion of human intestinal cells. Infect Immun 67(8):4171–4182
Konkel ME, Samuelson DR, Eucker TP et al (2013) Invasion of epithelial cells by Campylobacter jejuni is independent of caveolae. Cell Commun Signal 11:100. doi:10.1186/1478-811X-11-100
Acknowledgements
We thank Christopher R. Gourley for critical review of this manuscript and Joanna Fragoso for technical assistance.
Research in the Konkel Laboratory is supported by funds from the United States Department of Agriculture, National Institute of Food and Agriculture (Award Number 2011-67015-30772).
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Negretti, N.M., Konkel, M.E. (2017). Methods to Study Campylobacter jejuni Adherence to and Invasion of Host Epithelial Cells. In: Butcher, J., Stintzi, A. (eds) Campylobacter jejuni. Methods in Molecular Biology, vol 1512. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6536-6_11
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DOI: https://doi.org/10.1007/978-1-4939-6536-6_11
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