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Probiotic features of Lactobacillus plantarum mutant strains

  • Applied genetics and molecular biotechnology
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Abstract

In this study, the probiotic potential of Lactobacillus plantarum wild-type and derivative mutant strains was investigated. Bacterial survival was evaluated in an in vitro system, simulating the transit along the human oro-gastro-intestinal tract. Interaction with human gut epithelial cells was studied by assessing bacterial adhesive ability to Caco-2 cells and induction of genes involved in innate immunity. L. plantarum strains were resistant to the combined stress at the various steps of the simulated gastrointestinal tract. Major decreases in the viability of L. plantarum cells were observed mainly under drastic acidic conditions (pH ≤ 2.0) of the gastric compartment. Abiotic stresses associated to small intestine poorly affected bacterial viability. All the bacterial strains significantly adhered to Caco-2 cells, with the ΔctsR mutant strain exhibiting the highest adhesion. Induction of immune-related genes resulted higher upon incubation with heat-inactivated bacteria rather than with live ones. For specific genes, a differential transcriptional pattern was observed upon stimulation with different L. plantarum strains, evidencing a possible role of the knocked out bacterial genes in the modulation of host cell response. In particular, cells from Δhsp18.55 and ΔftsH mutants strongly triggered immune defence genes. Our study highlights the relevance of microbial genetic background in host–probiotic interaction and might contribute to identify candidate bacterial genes and molecules involved in probiosis.

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References

  • Ahrné S, Nobaek S, Jeppsson B, AdlerberthI WA, Molin G (1998) The normal Lactobacillus flora of healthy human rectal and oral mucosa. J Appl Microbiol 85:88–94

    Article  Google Scholar 

  • Akira S, Uematsu S, Takeuchi O (2006) Pathogen recognition and innate immunity. Cell 124:783–801

    Article  CAS  Google Scholar 

  • Bloksma N, de Heer E, van Dijk H, Willers JM (1979) Adjuvanticity of lactobacilli. I. Differential effects of viable and killed bacteria. Clin Exp Immunol 37:367–375

    CAS  Google Scholar 

  • Borchers AT, Selmi C, Meyers FJ, Keen CL, Gershwin ME (2009) Probiotics and immunity. J Gastroenterol 44:26–46

    Article  Google Scholar 

  • Bove P, Capozzi V, Garofalo C, Rieu A, Spano G, Fiocco D (2012) Inactivation of the ftsH gene of Lactobacillus plantarum WCFS1: effects on growth, stress tolerance, cell surface properties and biofilm formation. Microbiol Res 167:187–193

    Google Scholar 

  • Capozzi V, Weidmann S, Fiocco D, Rieu A, Hols P, Guzzo J, Spano G (2011) Inactivation of a small heat shock protein affects cell morphology and membrane fluidity in Lactobacillus plantarum WCFS1. Res Microbiol 162:419–425

    Article  CAS  Google Scholar 

  • Chr. Hansen (2011) http://www.chrhansen.com/products/product_areas/probiotics_for_human_health/strains/la_5r.html. Accessed 20 November 2011

  • Claes IJ, Lebeer S, Shen C, Verhoeven TL, Dilissen E, De Hertogh G, Bullens DM, Ceuppens JL, Van Assche G, Vermeire S, Rutgeerts P, Vanderleyden J, De Keersmaecker SC (2010) Impact of lipoteichoic acid modification on the performance of the probiotic Lactobacillus rhamnosus GG in experimental colitis. Clin Exp Immunol 162:306–314

    Article  CAS  Google Scholar 

  • de Vrese M, Schrezenmeir J (2008) Probiotics, prebiotics, and synbiotics. Adv Biochem Eng Biotechnol 111:1–66

    Google Scholar 

  • de Vries MC, Vaughan EE, Kleerebezem M, de Vos WM (2006) Lactobacillus plantarum—survival, functional and potential probiotic properties in the human intestinal tract. Int Dairy J 16:1018–1028

    Article  Google Scholar 

  • Derré I, Rapoport G, Msadek T (1999) CtsR, a novel regulator of stress and heat shock response, controls clp and molecular chaperone gene expression in Gram-positive bacteria. Mol Microbiol 31:117–131

    Article  Google Scholar 

  • Eckburg PB, Bik EM, Bernstein CN, Purdom E, Dethlefsen L, Sargent M, Gill SR, Nelson KE, Relman DA (2005) Diversity of the human intestinal microbial flora. Science 308:1635–1638

    Article  Google Scholar 

  • FAO/WHO (2002) ftp://ftp.fao.org/es/esn/food/wgreport2.pdf. Accessed 20 November 2011

  • Fernández de Palencia P, López P, Corbí AL, Peláez C, Requena T (2008) Probiotic strains: survival under simulated gastrointestinal conditions, in vitro adhesion to Caco-2 cells and effect on cytokine secretion. Eur Food Res Technol 227:1475–1484

    Article  Google Scholar 

  • Fiocco D, Collins M, Muscariello L, Hols P, Kleerebezem M, Msadek T, Spano G (2009) The Lactobacillus plantarum ftsH gene is a novel member of the CtsR stress response regulon. J Bacteriol 191:1688–1694

    Article  CAS  Google Scholar 

  • Fiocco D, Capozzi V, Collins M, Gallone A, Hols P, Guzzo J, Weidmann S, Rieu A, Msadek T, Spano G (2010) Characterization of the CtsR stress response regulon in Lactobacillus plantarum. J Bacteriol 192:896–900

    Article  CAS  Google Scholar 

  • Grangette C, Nutten S, Palumbo E, Morath S, Hermann C, Dewulf J, Pot B, Hartung T, Hols P, Mercenier A (2005) Enhanced antiinflammatory capacity of a Lactobacillus plantarum mutant synthesizing modified teichoic acids. Proc Natl Acad Sci USA 102:10321–10326

    Article  CAS  Google Scholar 

  • Guarner F, Malagelada JR (2003) Gut flora in health and disease. Lancet 361:512–519

    Article  Google Scholar 

  • Hooper LV, Gordon JI (2001) Commensal host–bacterial relationships in the gut. Science 292:1115–1118

    Article  CAS  Google Scholar 

  • Huang Y, Adams MC (2004) In vitro assessment of the upper gastrointestinal tolerance of potential probiotic dairy propionibacteria. Int J Food Microbiol 91:253–260

    Article  Google Scholar 

  • Isolauri E (2001) Probiotics in human disease. Am J Clin Nutr 73:1142–1146

    Google Scholar 

  • Isolauri E, Salminen S, Ouwehand AC (2004) Microbial–gut interactions in health and disease. Probiotics. Best Pract Res Clin Gastroenterol 18:299–313

    Article  Google Scholar 

  • Ito K, Akiyama Y (2005) Cellular functions, mechanism of action, andregulation of FtsH protease. Annu Rev Microbiol 59:211–231

    Article  CAS  Google Scholar 

  • Jijon H, Backer J, Diaz H, Yeung H, Thiel D, McKaigney C, De Simone C, Madsen K (2004) DNA from probiotic bacteria modulates murine and human epithelial and immune function. Gastroenterology 126:1358–1373

    Article  CAS  Google Scholar 

  • Johansson ML, Molin G, Jeppsson B, Nobaek S, Ahrné S, Bengmark S (1993) Administration of different Lactobacillus strains in fermented oatmeal soup: in vivo colonisation of human intestinal mucosa and effect on the indigenous flora. Appl Environ Microbiol 59:15–20

    CAS  Google Scholar 

  • Kalliomäki M, Salminen S, Isolauri E (2008) Positive interactions with the microbiota: probiotics. Adv Exp Med Biol 635:57–66

    Article  Google Scholar 

  • Kataria J, Li N, Wynn JL, Neu J (2009) Probiotic microbes: do they need to be alive to be beneficial? Nutr Rev 67:546–550

    Article  Google Scholar 

  • Kim YG, Ohta T, Takahashi T, Kushiro A, Nomoto K, Yokokura T, Okada N, Danbara H (2006) Probiotic Lactobacillus casei activates innate immunity via NF-kappaB and p38 MAP kinase signaling pathways. Microbes Infect 8:994–1005

    Article  CAS  Google Scholar 

  • Kleerebezem M, Boekhorst J, van Kranenburg R, Molenaar D, Kuipers OP, Leer R, Tarchini R, Peters SA, Sandbrink HM, Fiers MW, Stiekema W, Lankhorst RM, Bron PA, Hoffer SM, Groot MN, Kerkhoven R, de Vries M, Ursing B, de Vos WM, Siezen RJ (2003) Complete genome sequence of Lactobacillus plantarum WCFS1. Proc Natl Acad Sci USA 100:1990–1995

    Article  CAS  Google Scholar 

  • Kleerebezem M, Vaughan EE (2009) Probiotic and gut lactobacilli and bifidobacteria: molecular approaches to study diversity and activity. Annu Rev Microbiol 63:269–290

    Article  CAS  Google Scholar 

  • Konstantinov SR, Smidt H, de Vos WM, Bruijns SC, Singh SK (2008) S layer protein A of Lactobacillus acidophilus NCFM regulates immature dendritic cell and T cell functions. Proc Natl Acad Sci USA 105:19474–1979

    Article  CAS  Google Scholar 

  • Kwon JH, Keates S, Bassani L, Mayer LF, Keates AC (2002) Colonic epithelial cells are a major site of macrophage inflammatory protein 3alpha (MIP-3alpha) production in normal colon and inflammatory bowel disease. Gut 51:818–826

    Article  CAS  Google Scholar 

  • Lebeer S, Vanderleyden J, De Keersmaecker SC (2010) Host interactions of probiotic bacterial surface molecules: comparison with commensals and pathogens. Nat Rev Microbiol 8:171–184

    Article  CAS  Google Scholar 

  • LeBlanc JG, del Carmen S, Miyoshi A, Azevedo V, Sesma F, Langella P, Bermúdez-Humarán LG, Watterlot L, Perdigon G, de Moreno de LeBlanc A (2011) Use of superoxide dismutase and catalase producing lactic acid bacteria in TNBS induced Crohn's disease in mice. J Biotechnol 151:287–293

    Article  CAS  Google Scholar 

  • Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods 25:402–408

    Article  CAS  Google Scholar 

  • Ljungh A, Wadström T (2006) Lactic acid bacteria as probiotics. Curr Issues Intest Microbiol 7:73–89

    CAS  Google Scholar 

  • Lo Curto A, Pitino I, Mandalari G, Dainty JR, Faulks RM, John Wickham MS (2011) Survival of probiotic lactobacilli in the upper gastrointestinal tract using an in vitro gastric model of digestion. Food Microbiol 28:1359–1366

    Article  Google Scholar 

  • Mack DR, Michail S, Wei S, McDougall L, Hollingsworth MA (1999) Probiotics inhibit enteropathogenic E. coli adherence in vitro by inducing intestinal mucin gene expression. Am J Physiol 276:G941–950

    CAS  Google Scholar 

  • Mainville I, Arcand Y, Farnworth ER (2005) A dynamic model that simulates the human upper gastrointestinal tract for the study of probiotics. Int J Food Microbiol 99:287–296

    Article  CAS  Google Scholar 

  • Marteau P, Minekus M, Havenaar R, Huis in’t Veld JH (1997) Survival of lactic acid bacteria in a dynamic model of the stomach and small intestine: validation and the effects of bile. J Dairy Sci 80:1031–1037

    Article  CAS  Google Scholar 

  • Meijerink M, van Hemert S, Taverne N, Wels M, de Vos P, Bron PA, Savelkoul HF, van Bilsen J, Kleerebezem M, Wells JM (2010) Identification of genetic loci in Lactobacillus plantarum that modulate the immune response of dendritic cells using comparative genome hybridization. PLoS One 5:e10632

    Article  Google Scholar 

  • Mohamadzadeh M, Pfeiler EA, Brown JB, Zadeh M, Gramarossa M, Managlia E, Bere P, Sarraj B, Khan MW, Pakanati KC, Ansari MJ, O'Flaherty S, Barrett T, Klaenhammer TR (2011) Regulation of induced colonic inflammation by Lactobacillus acidophilus deficient in lipoteichoic acid. Proc Natl Acad Sci USA 108:4623–4630

    Article  CAS  Google Scholar 

  • Molin G (2001) Probiotics in foods not containing milk or milk constituents, with special reference to Lactobacillus plantarum 299v. Am J Clin Nutr 73:380–385

    Google Scholar 

  • Morita H, He F, Fuse T, Ouwehand AC, Hashimoto H, Hosoda M, Mizumachi K, Kurisaki J (2002) Adhesion of lactic acid bacteria to caco-2 cells and their effect on cytokine secretion. Microbiol Immunol 46:293–297

    CAS  Google Scholar 

  • Ohland CL, MacNaughton WK (2010) Probiotic bacteria and intestinal epithelial barrier function. Am J Physiol Gastrointest Liver Physiol 298:G807–G819

    Article  CAS  Google Scholar 

  • O’Hara AM, Shanahan F (2006) The gut flora as a forgotten organ. EMBO reports 7:688–693

    Article  Google Scholar 

  • Ou G, Baranov V, Lundmark E, Hammarström S, Hammarström L (2009) Contribution of intestinal epithelial cells to innate immunity of the human gut—studies on polarized monolayers of colon carcinoma cells. Scand J Immunol 69:150–161

    Article  CAS  Google Scholar 

  • Ouwehand AC, Salminen S, Isolauri E (2002) Probiotics an overview of beneficial effects. Antonie van Leeuwenhoek 83:279–289

    Article  Google Scholar 

  • Petrof EO, Kojima K, Ropeleski MJ, Musch MW, Tao Y, De Simone C, Chang EB (2004) Probiotics inhibit nuclear factor-kB and induce heat shock proteins in colonic epithelial cells through proteasome inhibition. Gastroenterology 127:1474–1487

    Article  CAS  Google Scholar 

  • Pinto M, Robine-Leon S, Appay M, Kidinger M, Traidou N, Dussaulx E (1983) Enterocyte-like differentiation of the human colon carcinoma cell line, Caco-2 in culture. Biol Cell 47:323–330

    Google Scholar 

  • Ruiz PA, Hoffmann M, Szcesny S, Blaut M, Haller D (2005) Innate mechanisms for Bifidobacterium lactis to activate transient proinflammatory host responses in intestinal epithelial cells after the colonization of germ-free rats. Immunology 115:441–450

    Article  CAS  Google Scholar 

  • Schlee M, Harder J, Köten B, Stange EF, Wehkamp J, Fellermann K (2008) Probiotic lactobacilli and VSL#3 induce enterocyte beta-defensin 2. Clin Exp Immunol 151:528–535

    Article  CAS  Google Scholar 

  • Spano G, Beneduce L, Perrotta C, Massa S (2005) Cloning and characterization of the hsp 18.55 gene, a new member of the small heat shock genes family isolated from wine Lactobacillus plantarum. Res Microbiol 156:219–224

    Article  CAS  Google Scholar 

  • van Baarlen P, Troost FJ, van Hemert S, van der Meer C, de Vos WM, de Groot PJ, Hooiveld GJ, Brummer RJ, Kleerebezem M (2009) Differential NF-kappaB pathways induction by Lactobacillus plantarum in the duodenum of healthy humans correlating with immune tolerance. Proc Natl Acad Sci U S A 106:2371–2376

    Article  Google Scholar 

  • Watterlot L, Rochat T, Sokol H, Cherbuy C, Bouloufa I, Lefèvre F, Gratadoux JJ, Honvo-Hueto E, Chilmonczyk S, Blugeon S, Corthier G, Langella P, Bermúdez-Humarán LG (2010) Intragastric administration of a superoxide dismutase-producing recombinant Lactobacillus casei BL23 strain attenuates DSS colitis in mice. Int J Food Microbiol 144:35–41

    Article  CAS  Google Scholar 

  • Wehkamp J, Harder J, Wehkamp K, Wehkamp-von Meissner B, Schlee M, Enders C, Sonnenborn U, Nuding S, Bengmark S, Fellermann K, Schröder JM, Stange EF (2004) NF-kappaB- and AP-1-mediated induction of human beta defensin-2 in intestinal epithelial cells by Escherichia coli Nissle 1917: a novel effect of a probiotic bacterium. Infect Immun 72:5750–5758

    Article  CAS  Google Scholar 

  • Wong C, Ustunol Z (2006) Mode of inactivation of probiotic bacteria affects interleukin 6 and interleukin 8 production in human intestinal epithelial-like Caco-2 cells. J Food Prot 69:2285–2288

    CAS  Google Scholar 

  • Yan F, Cao H, Cover TL, Whitehead R, Washington MK, Polk DB (2007) Soluble proteins produced by probiotic bacteria regulate intestinal epithelial cell survival and growth. Gastroenterology 132:562–575

    Article  CAS  Google Scholar 

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Acknowledgments

P. Bove is a Ph.D. student supported by the University of Foggia. The authors wish to thank Dr. Matteo Landriscina for providing Caco-2 cells.

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Authors and Affiliations

  1. Department of Biomedical Sciences, University of Foggia, Via L. Pinto 1, 71122, Foggia, Italy

    Pasquale Bove & Daniela Fiocco

  2. Department of Basic Medical Sciences, University of Bari “Aldo Moro”, Bari, Italy

    Anna Gallone

  3. Department of Food Sciences, University of Foggia, Via Napoli 25, 71122, Foggia, Italy

    Pasquale Russo, Vittorio Capozzi & Giuseppe Spano

  4. Department of Production Sciences and Innovation in Mediterranean Agriculture and Food Systems (PrIME), University of Foggia, Foggia, Italy

    Marzia Albenzio

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  1. Pasquale Bove
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  2. Anna Gallone
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  3. Pasquale Russo
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  4. Vittorio Capozzi
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  5. Marzia Albenzio
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  6. Giuseppe Spano
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  7. Daniela Fiocco
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Correspondence to Giuseppe Spano.

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Bove, P., Gallone, A., Russo, P. et al. Probiotic features of Lactobacillus plantarum mutant strains. Appl Microbiol Biotechnol 96, 431–441 (2012). https://doi.org/10.1007/s00253-012-4031-2

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  • DOI: https://doi.org/10.1007/s00253-012-4031-2

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