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2015 | 62 | 4 | 697-706
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The microbiological, histological, immunological and molecular determinants of Helicobacter pylori infection in guinea pigs as a convenient animal model to study pathogenicity of these bacteria and the infection dependent immune response of the host

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Helicobacter pylori is an etiological agent of chronic gastritis, gastric and duodenal ulcers and gastric cancers. The use of an appropriate animal model for experimental studies on the pathogenesis of H. pylori infections is necessary due to the chronic character of such infections and difficulties in identifying their early stage in humans. The aim of this study was to develop a guinea pig model of H. pylori infection and identify its microbiological, histological, serological and molecular determinants. Guinea pigs were inoculated per os with H. pylori strains: CCUG 17874 or ATCC 700312, both producing vacuolating cytotoxin A (VacA) and cytotoxin associated gene A (CagA) protein, suspended in Brucella broth with fetal calf serum (FCS) and Skirrow supplement of antibiotics. To determine H. pylori colonization, 7 and 28 days after the challenge, a panel of diagnostic methods was used. It included culturing of microorganisms from the gastric tissue, histopathological analysis of gastric sections, stained by Mayer,s haematoxylin and eosin to assess inflammatory response, by Giemsa as well as Warthin-Starry silver staining to visualise Helicobacter-like organisms (HLO) and with anti-Ki-67 antigen to assess epithelial cell proliferation. H. pylori infection was also confirmed by polymerase chain reactions (PCR) for detection in gastric tissue of ureC and cagA genes and by serological assessment of H. pylori antigens in faeces. This study showed the usefulness of microbiological, histological, immunological and molecular methods for the detection of persistent H. pylori infections in guinea pigs, which could be an appropriate model for studying H. pylori pathogenesis and the related immune response against these microbes.
Physical description
  • Division of Gastroimmunology, Department of Immunology and Infectious Biology, Institute of Microbiology, Biotechnology and Immunology, Faculty of Biology and Environmental Protection, University of Lodz, Łódź, Poland
  • Division of Gastroimmunology, Department of Immunology and Infectious Biology, Institute of Microbiology, Biotechnology and Immunology, Faculty of Biology and Environmental Protection, University of Lodz, Łódź, Poland
  • Division of Gastroimmunology, Department of Immunology and Infectious Biology, Institute of Microbiology, Biotechnology and Immunology, Faculty of Biology and Environmental Protection, University of Lodz, Łódź, Poland
  • Division of Gastroimmunology, Department of Immunology and Infectious Biology, Institute of Microbiology, Biotechnology and Immunology, Faculty of Biology and Environmental Protection, University of Lodz, Łódź, Poland
  • Department of Pathomorphology, Medical University of Lodz, Łódź, Poland
  • Department of Pathomorphology, Hospital under the name of K. Bieganski, Łódź, Poland
  • Division of Gastroimmunology, Department of Immunology and Infectious Biology, Institute of Microbiology, Biotechnology and Immunology, Faculty of Biology and Environmental Protection, University of Lodz, Łódź, Poland
  • Andersen LP, Rasmussen L (2009) Helicobacter pylori-coccoid forms and biofilm formation. FEMS Immunol Med Microbiol 56: 112-115. Doi: 10.1111/j.1574-695X.
  • Antczak DF (1982) Structure and function of the major histocompatibility complex in domestic animals. J Am Vet Med Assoc 181: 1030-1036.
  • Amirhooshang A, Ramin A, Ehsan A, Mansour R, Shahram B (2014) High frequency of Helicobacter pylori DNA in drinking water in Kermanshah, Iran, during June-November 2012. J Water Health 12: 504-512.
  • Ahmed KS, Khan AA, Ahmed I, Tiwari SK, Habeeb A, Ahi JD, Abid Z, Ahmed N, Habibullah CM (2007) Impact of household hygiene and water source on the prevalence and transmission of Helicobacter pylori: a South Indian perspective. Singapore Med J 48: 543-549.
  • Baudron CR, Franceschi F, Salles N, Gasbarrini A (2013) Extragastric diseases and Helicobacter pylori. Helicobacter 18 (Suppl 1): 44-51. DIO: 10.1111/hel.12077.
  • Bielanski W, Konturek SJ (1996) New approach to 13C urea breath test capsule-based modification with low dose of 13C urea in the diagnosis of Helicobacter pylori infection. J Physiol Pharmacol 47: 545-553.
  • Bickley J, Owen RJ, Fraser AG, Pounder RE (1993) Evaluation of the polymerase chain reaction for detecting the urease C gene of Helicobacter pylori in gastric biopsy samples and dental plaque. J Med Microbiol 39: 338-344.
  • Bitter-Suermann D, Hoffman T, Burger R, Hadding U (1981) Linkage of total deficiency of the second component (C2) of the complement system and of genetic C2 polymorphism to the major histocompatibility comples of the guinea pig. J Immunol 127: 608-612.
  • Bode G, Mauch F, Malfetheiner P (1993) The coccoid forms of Helicobacter pylori. Criteria for their viability. Epidemiol and Infect 111: 483-490.
  • Campbell EM, Proudfoot AE, Yoshimura T, Allet B, Wells TN, White AM, Westwick J, Watson ML (1997) Recombinant guinea pig and human RANTES activate macrophages but not eosinophils in the guinea pig. J Immunol 159: 1482-1489.
  • Chisholm SA, Owen RJ, Teare EL, Saverymuttu S (2001) PCR-based diagnosis of Helicobacter pylori infection and real-time determinantion of claritromycin resistance directly from human gastric biopsy samples. J Clin Microbiol 39: 1217-1220.
  • Chmiela M, Michetti P (2006) Inflammation, immunity, vaccines for Helicobacter pylori infection. Helicobacter 11: 21-26.
  • Chmiela M, Miszczyk E, Rudnicka K (2014) Structural modifications of Helicobacter pylori lipopolysaccharide: an idea for how to live in peace. World J Gastroenterol 20: 9882-9887.
  • Chmiela M, Gajewski A, Rudnicka K (2015) Helicobacter pylori vs coronary heart disease-searching for connections. World J Cardiol 7: 187-203.
  • Claman HN (1972) Corticosteroids and lymphoid cells. N Engl J Med 287: 388-397.
  • Covacci A, Censini S, Bugnoli M, Petracca R, Burroni D, Macchia G, Massone A, Papini E, Xiang Z, Figura N, Rappuoli R (1993) Molecular characterization of the 128-kDa immunodominant antigen of Helicobacter pylori associated with cytotoxicity and duodenal ulcer. Proc Natl Acad Sci USA 90: 5791-5795.
  • Deguchi R, Matsushima M, Suzuki T, Mine T, Fukuda R, Nishina M, Ozawa H, Takagi A (2009) Comparison of a monoclonal with polyclonal antibody-based enzyme immunoassay stool test in diagnosing Helicobacter pylori infection after eradication therapy. J Gastroenterol 44: 713-716.
  • D'Erchia AM, Gissi C, Pesole G, Saccone C, Arnason U (1996) The guinea-pig is not a rodent. Nature 381: 597-600.
  • Dixon MF (1991) Helicobacter pylori and peptic ulceration: histological aspects. J Gastroenterol Hepatol 6: 125-130.
  • Fujimura S, Kato S, Watanabe A (2008) Water source as a Helicobacter pylori transmission route: a 3-year follow-up study of Japanese children living in a unique district. J Med Microbiol 57 (Pt7): 909-910.
  • Ganguly R, Durieux MF, Waldman RH (1976) Macrophage function in vitamin C-deficient guinea pigs. Am J Clin Nutr 29: 762-765.
  • Grande R, Di Campli E, Di Bartolomeo S, Verginelli F, Di Giulio M, Baffoni M, Bessa LJ, Cellini L (2012) Helicobacter pylori biofilm: a protective environment for bacterial recombination. J Appl Microbiol 113: 669-76.
  • Grebowska A, Moran AP, Bielanski W, Matusiak A, Rechcinski T, Rudnicka K, Baranowska A, Rudnicka W, Chmiela M (2010) Helicobacter pylori lipopolysaccharide activity in human peripheral blood mononuclear leukocyte cultures. J Physiol Pharmacol 61: 437-442.
  • Hiromatsu K, Dascher CC, Sugita M, Gingrich-Baker C, Behar SM, LeClair KP, Brenner MB, Porcelli SA (2002) Characterization of guinea-pig group 1 CD1 proteins. Immunology 106: 159-172.
  • Jeevan A, McFarland CT, Yoshimura T, Skwor T, Cho H, Lasco T, McMurray DN (2006) Production and characterization of guinea pig recombinant gamma interferon and its effect on macrophage activation. Infect Immun 74: 213-224.
  • Kuo CH, Chen YH, Goh KL, Chang LL (2014) Helicobacter pylori and systemic diseases. Gastroenterol Res Pract 2014: 358494.
  • Lee A, O'Rourke J, De Ungria MC, Robertson B, Daskalopoulos G, Dixon MF (1997) A standardized mouse model of Helicobacter pylori infection: introducing the Sydney strain. Gastroenterology 112: 1386-1397.
  • Lee JY, Kim N (2015) Diagnosis of Helicobacter pylori by invasive test: histology. Ann Transl Med 3: 10.
  • Li Ch, Ha T, Chi DS, Ferguson DA, Jiang Ch, Laffan JJ, Thomas E (1997) Differentiation of Helicobacter pylori strains directly from gastric biopsy specimens by PCR-based restriction fragment length polymorphism analysis without culture. J Clin Microbiol 35: 3021-3025.
  • Lina TT, Alzahrani S, Gonzalez J, Pinchuk IV, Beswick EJ, Reyes V (2014) Immune evasion strategies used by Helicobacter pylori. World J Gastroenterol 20: 12753-12766.
  • Marshall BJ, Warren JR (1984) Unidentified curved bacilli in the stomach of patients with gastritis and peptic ulceration. Lancet 1: 1311-1315.
  • Megraud F, Broutet N (2000) Review article: have we found the source of Helicobacter pylori? Aliment Pharmacol Ther 14 (Suppl 3): 7-12.
  • Megraud F (2012) The challenge of Helicobacter pylori resistance to antibiotics: the comeback of bismuth-based quadruple therapy. Therap Adv Gastroenterol 5: 103-109.
  • Malfertheiner P, Megraud F, O'Morain CA, Atherton J, Axon AT, Bazzoli F, Gensini GF, Gisbert JP, Graham DY, Rokkas T, El-Omar EM, Kuipers EJ, European Helicobacter Study Group (2012) Management of Helicobacter pylori infection-the Maastricht IV/Florence Consensus Report. Gut 61: 646-664.
  • McColl KE (2010) Clinical practice. Helicobacter pylori infection. N Engl J Med 362: 1597-1604.
  • McGee DJ, May CA, Garner RM, Himpsl JM, Mobley HL (1999) Isolation of Helicobacter pylori genes that modulate urease activity. J Bacteriol 181: 2477-2484.
  • McMurray DN (1994) Disease model: pulmonary tuberculosis. Trends Mol Med 7: 135.
  • Meurs H, Santing RE, Remie R, van der Mark TW, Westerhof FJ, Zuidhof AB, Bos IS, Zaagsma J (2006) A guinea pig model of acute and chronic asthma using permanently instrumented and unrestrained animals. Nat Protocols 1: 840-847.
  • Michalkiewicz J, Helmin-Basa A, Grzywa R, Czerwionka-Szaflarska M, Szaflarska-Poplawska A, Mierzwa G, Marszalek A, Bodnar M, Nowak M, Dzierzanowska-Fangrat K (2015) Innate immunity components and cytokines in gastric mucosa in children with Helicobacter pylori infection. Mediators Inflamm 2015: 176726.
  • Odenbreit S (2005) Adherence properties of Helicobacter pylori: Impact on pathogenesis and adaptation to the host. Int J Med Microbiol 295: 317-324.
  • O'Rourke JL, Lee A (2003) Animal models of Helicobacter pylori infection and disease. Microbes Infect 5: 741-748.
  • Padilla-Carlin DJ, McMurray DN, Hickey AJ (2008) The guinea pig as a model of infectious diseases. Comp Med 58: 324-340.
  • Peek RM (2008) Helicobacter pylori infection and disease: from humans to animal models. Disease Models and Mechanisms 1: 50-55.
  • Price AB (1991) The Sydney System: histological division. J Gastroenterol Hepatol 6: 209-222.
  • Rechciński T, Chmiela M, Małecka-Panas E, Płaneta-Małecka I, Rudnicka W (1997) Serological indicators of Helicobacter pylori infection in adult dyspeptic patients and healthy blood donors. Microbiol Immunol 41: 387-393.
  • Rijpkema SG, Durrani Z, Beavan G, Gibson JR, Luck J, Owen RJ, Auda GR (2001) Analysis of host responses of guinea pigs during Helicobacter pylori infection. FEMS Immunol Med Microbiol 30: 151-156.
  • Rudnicka K, Miszczyk E, Matusiak A, Walencka M, Moran AP, Rudnicka W, Chmiela M (2015) Helicobacter pylori-driven modulation of NK cell expansion, intracellular cytokine expression and cytotoxic activity. Innate Immun 21: 127-139.
  • Shiratori I, Matsumoto M, Tsuji S, Nomura M, Toyoshima K, Seya T (2001) Molecular cloning and functional characterization of guinea pig IL-12. Int Immunol 13: 1129-1139.
  • Sjunnesson H, Sturegard E, Hynes S, Willen R, Feinstein R, Wadstrom T (2003) Five month persistence of Helicobacter pylori infection in guinea pigs. APMIS 111: 634-642.
  • Stevenson TH, Castillo A, Lucia LM, Acuff GR (2000) Growth of Helicobacter pylori in various liquid and plating media. Lett Appl Microbiol 30: 192-196.
  • Stingl K, De Reuse H (2005) Staying alive overdosed: How does Helicobacter pylori control urease activity? Int J Med Microbiol 295: 307-315.
  • Stolte M, Meining A (2001) The updated Sydney system classification and grading of gastritis as the basis diagnosis and treatment. Can J Gastroenterol 15: 591-598.
  • Sturegard E, Sjunnesson H, Ho B, Willen R, Aleljung P, NG HC, Wadstrom T (1998) Severe gastritis in guinea pigs infected with Helicobacter pylori. J Med Microbiol 47: 1123-1129.
  • Takahashi M, Jeevan A, Sawant K, McMurray DN, Yoshimura T (2007) Cloning and characterization of guinea pig CXCR1. Mol Immunol 44: 878-888.
  • Whary MT, Fox JG (2004) Natural and experimental Helicobacter infection. Comp Med 54: 128-158.
  • Wicher V, Scarozza AM, Ramsingh AL, Wicher K (1998) Cytokine gene expression in skin of susceptible guinea pigs infected with Treponema pallidum. Immunology 95: 242-247.
  • Wisniewska M, Nilsson HO, Bak-Romaniszyn L, Rechcinski T, Bielanski W, Planeta-Malecka I, Plonka M, Konturek S, Wadstrom T, Rudnicka W, Chmiela M (2002) Detection of specific Helicobacter pylori DNA and antigens in stool samples in dyspeptic patients and health subjects. Microbiol Immunol 46: 657-665.
  • Wroblewski LE, Peek RM Jr, Wilson KT (2010) Helicobacter pylori and gastric cancer: factors that modulate disease risk. Clin Microbiol Rev 23: 713-739.
  • Wu W, Yang Y, Sun G (2012) Recent insights into antibiotic resistance in Helicobacter pylori eradication. Gastroenterol Res Pract 2012: 723183.
  • Wu WM, Yang YS, Peng LH (2014) Microbiota in the stomach: New insights. J Dig Dis 15: 54-61.
  • Vaira D, Holton J, Oderda G, Taylor D, Turner R, Colecchia A, Gandolfi L (1991) Transmission and sources of H. pylori. In Helicobacter pylori 1990, Menge M, Gregor M, Tytgat NJ, Marshall BJ, Mc Nulty CAM eds. Springer-Verlag 1991.
  • Varbanova M, Frauenschlager K, Malfertheiner P (2014) Chronic gastritis - an update. Best Pract Res Clin Gastroenterol 28: 1031-1042.
  • Vale FF, Vitor JM (2010) Transmission pathway of Helicobacter pylori: does food play a role in rural and urban areas? Int J Food Microbiol 138: 1-12.
  • Vogiatzi P, Cassone M, Luzzi I, Lucchetti C, Otvos L Jr, Giordano A (2007) Helicobacter pylori as a class I carcinogen: physiopathology and management strategies. J Cell Biochem 102: 264-273.
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