1887

Journal of Medical Microbiology logo

Volume 73, Issue 1

Genomic and vaccine preclinical studies reveal a novel mouse-adapted model for the hpEastAsia genotype in Southeast Asia Open Access

Abstract

infection is a major global health concern, linked to the development of various gastrointestinal diseases, including gastric cancer. To study the pathogenesis of and develop effective intervention strategies, appropriate animal pathogen models that closely mimic human infection are essential.

This study focuses on the understudied hpEastAsia genotype in Southeast Asia, a region marked by a high infection rate. No mouse-adapted model strains has been reported previously. Moreover, it recognizes the urgent requirement for vaccines in developing countries, where overuse of antimicrobials is fuelling the emergence of resistance.

This study aims to establish a novel mouse-adapted model specific to the hpEastAsia genotype prevalent in Southeast Asia, focusing on comparative genomic and histopathological analysis of pathogens coupled with vaccine preclinical studies.

We collected and sequenced the whole genome of clinical strains of from infected patients in Vietnam and performed comparative genomic analyses of strains in Southeast Asia. In parallel, we conducted preclinical studies to assess the pathogenicity of the mouse-adapted strain and the protective effect of a new spore-vectored vaccine candidate on male Mlac:ICR mice and the host immune response in a female C57BL/6 mouse model.

Genome sequencing and comparison revealed unique and common genetic signatures, antimicrobial resistance genes and virulence factors in strains HP22 and HP34; and supported clarithromycin-resistant HP34 as a representation of the hpEastAsia genotype in Vietnam and Southeast Asia. HP34-infected mice exhibited gastric inflammation, epithelial erosion and dysplastic changes that closely resembled the pathology observed in human infection. Furthermore, comprehensive immunological characterization demonstrated a robust host immune response, including both mucosal and systemic immune responses. Oral vaccination with candidate vaccine formulations elicited a significant reduction in bacterial colonization in the model.

Our findings demonstrate the successful development of a novel mouse-adapted model for the hpEastAsia genotype in Vietnam and Southeast Asia. Our research highlights the distinctive genotype and pathogenicity of clinical strains in the region, laying the foundation for targeted interventions to address this global health burden.

  • Received:
  • Accepted:
  • Published Online:
Keyword(s): gastric cancer , gastritis , genomics , Helicobacter pylori , hpEastAsia and spore vectored vaccine
Funding
This study was supported by the:
  • Ministry of Science and Technology (Award NDT.79.GB/20)
    • Principle Award Recipient: VanDuy Nguyen
  • Medical Research Council (Award MR/R026262/1)
    • Principle Award Recipient: SimonCutting
© 2024 Crown Copyright
  • This is an open-access article distributed under the terms of the Creative Commons Attribution License. This article was made open access via a Publish and Read agreement between the Microbiology Society and the corresponding author’s institution.
Loading

Article metrics loading...

/content/journal/jmm/10.1099/jmm.0.001786
2024-01-18
2024-04-28
Loading full text...

Full text loading...

/deliver/fulltext/jmm/73/1/jmm001786.html?itemId=/content/journal/jmm/10.1099/jmm.0.001786&mimeType=html&fmt=ahah

References

  1. de Martel C, Georges D, Bray F, Ferlay J, Clifford GM. Global burden of cancer attributable to infections in 2018: a worldwide incidence analysis. Lancet Glob Health 2020; 8:e180–e190 [View Article] [PubMed]
     [Google Scholar]
  2. Plummer M, Franceschi S, Vignat J, Forman D, de Martel C. Global burden of gastric cancer attributable to Helicobacter pylori. Int J Cancer 2015; 136:487–490 [View Article] [PubMed]
     [Google Scholar]
  3. Elbehiry A, Marzouk E, Aldubaib M, Abalkhail A, Anagreyyah S et al. Helicobacter pylori Infection: current status and future prospects on diagnostic, therapeutic and control challenges. Antibiotics 2023; 12:191 [View Article] [PubMed]
     [Google Scholar]
  4. Burucoa C, Axon A. Epidemiology of Helicobacter pylori infection. Helicobacter 2017; 22:e12403 [View Article] [PubMed]
     [Google Scholar]
  5. Pucułek M, Machlowska J, Wierzbicki R, Baj J, Maciejewski R et al. Helicobacter pylori associated factors in the development of gastric cancer with special reference to the early-onset subtype. Oncotarget 2018; 9:31146–31162 [View Article] [PubMed]
     [Google Scholar]
  6. Sharndama HC, Mba IE. Helicobacter pylori: an up-to-date overview on the virulence and pathogenesis mechanisms. Braz J Microbiol 2022; 53:1–18 [View Article] [PubMed]
     [Google Scholar]
  7. Falush D, Wirth T, Linz B, Pritchard JK, Stephens M et al. Traces of human migrations in Helicobacter pylori populations. Science 2003; 299:1582–1585 [View Article] [PubMed]
     [Google Scholar]
  8. GBD 2017 Stomach Cancer Collaborators The global, regional, and national burden of stomach cancer in 195 countries, 1990-2017: a systematic analysis for the Global Burden of Disease study 2017. In The Lancet. Gastroenterology & Hepatology vol 5 2017 pp 42–54 [View Article]
     [Google Scholar]
  9. Kawai M, Furuta Y, Yahara K, Tsuru T, Oshima K et al. Evolution in an oncogenic bacterial species with extreme genome plasticity: Helicobacter pylori East Asian genomes. BMC Microbiol 2011; 11:104 [View Article] [PubMed]
     [Google Scholar]
  10. You Y, Thorell K, He L, Yahara K, Yamaoka Y et al. Genomic differentiation within East Asian Helicobacter pylori. Microb Genom 2022; 8:000676 [View Article] [PubMed]
     [Google Scholar]
  11. Ansari S, Yamaoka Y. Animal models and Helicobacter pylori infection. J Clin Med 2022; 11:3141 [View Article]
     [Google Scholar]
  12. Ghosh N, Ghosh P, Kesh K, Mukhopadhyay AK, Swarnakar S. Attenuation of Helicobacter pylori-induced gastric inflammation by prior cag − strain (AM1) infection in C57BL/6 mice. Gut Pathog 2017; 9:14 [View Article]
     [Google Scholar]
  13. Patra R, Chattopadhyay S, De R, Ghosh P, Ganguly M et al. Multiple infection and microdiversity among Helicobacter pylori isolates in a single host in India. PLoS One 2012; 7:e43370 [View Article] [PubMed]
     [Google Scholar]
  14. Dey TK, Karmakar BC, Sarkar A, Paul S, Mukhopadhyay AK. A Mouse Model of Helicobacter pylori Infection. In Smith SM. eds Helicobacter PyloriMethods in Molecular Biology vol 2283 New York, NY: Humana; 2021 pp 131–151 [View Article] [PubMed]
     [Google Scholar]
  15. Lee A. Animal models of Helicobacter infection. Mol Med Today 1999; 5:500–501 [View Article] [PubMed]
     [Google Scholar]
  16. Katsande PM, Nguyen VD, Nguyen TLP, Nguyen TKC, Mills G et al. Prophylactic immunization to Helicobacter pylori infection using spore vectored vaccines. Helicobacter 2023; 28:e12997 [View Article] [PubMed]
     [Google Scholar]
  17. Berthenet E, Yahara K, Thorell K, Pascoe B, Meric G et al. A GWAS on Helicobacter pylori strains points to genetic variants associated with gastric cancer risk. BMC Biol 2018; 16:84 [View Article] [PubMed]
     [Google Scholar]
  18. Muñoz-Ramirez ZY, Pascoe B, Mendez-Tenorio A, Mourkas E, Sandoval-Motta S et al. A 500-year tale of co-evolution, adaptation, and virulence: Helicobacter pylori in the Americas. ISME J 2021; 15:78–92 [View Article] [PubMed]
     [Google Scholar]
  19. Zeigler DR, Prágai Z, Rodriguez S, Chevreux B, Muffler A et al. The origins of 168, W23, and other Bacillus subtilis legacy strains. J Bacteriol 2008; 190:6983–6995 [View Article] [PubMed]
     [Google Scholar]
  20. Harwood CR, Archibald AR. Growth, maintenance and general techniques. In Harwood CR, Cutting SM. eds Molecular Biological Methods for Bacillus vol 1990 Chichester, UK: John Wiley & Sons Ltd; 1990 pp 1–26
     [Google Scholar]
  21. Nicholson WL, Setlow P. Sporulation, Germination and Outgrowth. In Harwood CR, Cutting SM. eds Molecular Biological Methods for Bacillus Chichester, UK: John Wiley & Sons Ltd; 1990 pp 391–450
     [Google Scholar]
  22. Andrews S. FastQC: A Quality Control Tool for High Throughput Sequence Data Available online; 2010 http://www.bioinformatics.babraham.ac.uk/projects/fastqc
  23. Lanfear R, Schalamun M, Kainer D, Wang W, Schwessinger B. MinIONQC: fast and simple quality control for MinION sequencing data. Bioinformatics 2019; 35:523–525 [View Article] [PubMed]
     [Google Scholar]
  24. Bolger AM, Lohse M, Usadel B. Trimmomatic: a flexible trimmer for Illumina sequence data. Bioinformatics 2014; 30:2114–2120 [View Article] [PubMed]
     [Google Scholar]
  25. Lee S, Nguyen LT, Hayes BJ, Ross EM. Prowler: a novel trimming algorithm for Oxford Nanopore sequence data. Bioinformatics 2021; 37:3936–3937 [View Article] [PubMed]
     [Google Scholar]
  26. Wick RR, Judd LM, Gorrie CL, Holt KE, Phillippy AM. Unicycler: resolving bacterial genome assemblies from short and long sequencing reads. PLOS Comput Biol 2017; 13:e1005595 [View Article] [PubMed]
     [Google Scholar]
  27. Tatusova T, DiCuccio M, Badretdin A, Chetvernin V, Nawrocki EP et al. NCBI prokaryotic genome annotation pipeline. Nucleic Acids Res 2016; 44:6614–6624 [View Article] [PubMed]
     [Google Scholar]
  28. Cumsille A, Durán RE, Rodríguez-Delherbe A, Saona-Urmeneta V, Cámara B et al. GenoVi, an open-source automated circular genome visualizer for bacteria and archaea. PLoS Comput Biol 2023; 19:e1010998 [View Article] [PubMed]
     [Google Scholar]
  29. Liu Y, Wang S, Yang F, Chi W, Ding L et al. Antimicrobial resistance patterns and genetic elements associated with the antibiotic resistance of Helicobacter pylori strains from Shanghai. Gut Pathog 2022; 14:14 [View Article] [PubMed]
     [Google Scholar]
  30. Jiang X, Xu Z, Zhang T, Li Y, Li W et al. Whole-genome-based Helicobacter pylori geographic surveillance: a visualized and expandable webtool. Front Microbiol 2021; 12:687259 [View Article] [PubMed]
     [Google Scholar]
  31. Nguyen TKC, Pham TL, Nguyen TC, Pham TT, Nguyen VD. Environmentally responsible bioengineering for spore surface expression of Helicobacter pylori antigen. J Pure Appl Microbiol 2023; 17:1605–1611 [View Article]
     [Google Scholar]
  32. Dyer V, Brüggemann H, Sörensen M, Kühl AA, Hoffman K et al. Genomic features of the Helicobacter pylori strain PMSS1 and its virulence attributes as deduced from its in vivo colonisation patterns. Mol Microbiol 2018; 110:761–776 [View Article] [PubMed]
     [Google Scholar]
  33. Nguyen TH, Ho TTM, Nguyen-Hoang T-P, Qumar S, Pham TTD et al. The endemic Helicobacter pylori population in Southern Vietnam has both South East Asian and European origins. Gut Pathog 2021; 13:57 [View Article] [PubMed]
     [Google Scholar]
  34. Phuc BH, Tuan VP, Binh TT, Tung PH, Tri TD et al. Comparative genomics of two Vietnamese Helicobacter pylori strains, CHC155 from a non-cardia gastric cancer patient and VN1291 from a duodenal ulcer patient. Sci Rep 2023; 13:8869 [View Article] [PubMed]
     [Google Scholar]
  35. Lee A, O’Rourke J, De Ungria MC, Robertson B, Daskalopoulos G et al. A standardized mouse model of Helicobacter pylori infection: introducing the Sydney strain. Gastroenterology 1997; 112:1386–1397 [View Article] [PubMed]
     [Google Scholar]
  36. Crabtree JE, Ferrero RL, Kusters JG. The mouse colonizing Helicobacter pylori strain SS1 may lack a functional cag pathogenicity island. Helicobacter 2002; 7:139–140 [View Article] [PubMed]
     [Google Scholar]
  37. Israel DA, Salama N, Arnold CN, Moss SF, Ando T et al. Helicobacter pylori strain-specific differences in genetic content, identified by microarray, influence host inflammatory responses. J Clin Invest 2001; 107:611–620 [View Article] [PubMed]
     [Google Scholar]
  38. Fox JG, Rogers AB, Ihrig M, Taylor NS, Whary MT et al. Helicobacter pylori-associated gastric cancer in INS-GAS mice is gender specific. Cancer Res 2003; 63:942–950 [PubMed]
     [Google Scholar]
  39. Fox JG, Wang TC, Rogers AB, Poutahidis T, Ge Z et al. Host and microbial constituents influence Helicobacter pylori-induced cancer in a murine model of hypergastrinemia. Gastroenterology 2003; 124:1879–1890 [View Article] [PubMed]
     [Google Scholar]
  40. Hansen LM, Dekalb DJ, Cai LP, Solnick JV. Identification of pathogenicity island genes associated with loss of type IV secretion function during murine Infection with Helicobacter pylori. Infect Immun 2020; 88:e00801-19 [View Article]
     [Google Scholar]
  41. Thi Huyen Trang T, Thanh Binh T, Yamaoka Y. Relationship between vacA types and development of gastroduodenal diseases. Toxins 2016; 8:182 [View Article] [PubMed]
     [Google Scholar]
  42. Wang X, Willén R, Svensson M, Ljungh A, Wadström T. Two-year follow-up of Helicobacter pylori infection in C57BL/6 and Balb/cA mice. APMIS 2003; 111:514–522 [View Article] [PubMed]
     [Google Scholar]
  43. Takata T, El-Omar E, Camorlinga M, Thompson SA, Minohara Y et al. Helicobacter pylori does not require Lewis X or Lewis Y expression to colonize C3H/HeJ mice. Infect Immun 2002; 70:3073–3079 [View Article] [PubMed]
     [Google Scholar]
  44. Ferrero RL, Thiberge JM, Huerre M, Labigne A. Immune responses of specific-pathogen-free mice to chronic Helicobacter pylori (strain SS1) infection. Infect Immun 1998; 66:1349–1355 [View Article] [PubMed]
     [Google Scholar]
  45. Garhart CA, Redline RW, Nedrud JG, Czinn SJ. Clearance of Helicobacter pylori infection and resolution of postimmunization gastritis in a kinetic study of prophylactically immunized mice. Infect Immun 2002; 70:3529–3538 [View Article] [PubMed]
     [Google Scholar]
  46. van Doorn NE, Namavar F, Sparrius M, Stoof J, van Rees EP et al. Helicobacter pylori-associated gastritis in mice is host and strain specific. Infect Immun 1999; 67:3040–3046 [View Article] [PubMed]
     [Google Scholar]
  47. Kodama M, Murakami K, Nishizono A, Fujioka T. Animal models for the study of Helicobacter-induced gastric carcinoma. J Infect Chemother 2004; 10:316–325 [View Article] [PubMed]
     [Google Scholar]
  48. Pritchard DM, Przemeck SMC. Review article: how useful are the rodent animal models of gastric adenocarcinoma?. Aliment Pharmacol Ther 2004; 19:841–859 [View Article] [PubMed]
     [Google Scholar]
  49. O’Rourke JL, Lee A. Animal models of Helicobacter pylori infection and disease. Microbes Infect 2003; 5:741–748 [View Article] [PubMed]
     [Google Scholar]
  50. Cai X, Stoicov C, Li H, Carlson J, Whary M et al. Overcoming fas-mediated apoptosis accelerates Helicobacter-induced gastric cancer in mice. Cancer Res 2005; 65:10912–10920 [View Article] [PubMed]
     [Google Scholar]
  51. Kuzushita N, Rogers AB, Monti NA, Whary MT, Park MJ et al. p27kip1 deficiency confers susceptibility to gastric carcinogenesis in Helicobacter pylori-infected mice. Gastroenterology 2005; 129:1544–1556 [View Article] [PubMed]
     [Google Scholar]
  52. Ohnishi N, Yuasa H, Tanaka S, Sawa H, Miura M et al. Transgenic expression of Helicobacter pylori CagA induces gastrointestinal and hematopoietic neoplasms in mouse. Proc Natl Acad Sci U S A 2008; 105:1003–1008 [View Article] [PubMed]
     [Google Scholar]
  53. Tu S, Bhagat G, Cui G, Takaishi S, Kurt-Jones EA et al. Overexpression of interleukin-1beta induces gastric inflammation and cancer and mobilizes myeloid-derived suppressor cells in mice. Cancer Cell 2008; 14:408–419 [View Article] [PubMed]
     [Google Scholar]
  54. Wang TC, Dangler CA, Chen D, Goldenring JR, Koh T et al. Synergistic interaction between hypergastrinemia and Helicobacter infection in a mouse model of gastric cancer. Gastroenterology 2000; 118:36–47 [View Article] [PubMed]
     [Google Scholar]
  55. Yamamoto T, Kita M, Ohno T, Iwakura Y, Sekikawa K et al. Role of tumor necrosis factor-alpha and interferon-gamma in Helicobacter pylori infection. Microbiol Immunol 2004; 48:647–654 [View Article] [PubMed]
     [Google Scholar]
  56. Radin JN, Gaddy JA, González-Rivera C, Loh JT, Algood HMS et al. Flagellar localization of a Helicobacter pylori autotransporter protein. mBio 2013; 4:e00613-12 [View Article] [PubMed]
     [Google Scholar]
  57. Sause WE, Castillo AR, Ottemann KM. The Helicobacter pylori autotransporter ImaA (HP0289) modulates the immune response and contributes to host colonization. Infect Immun 2012; 80:2286–2296 [View Article] [PubMed]
     [Google Scholar]
  58. Xie W, Zhao W, Zou Z, Kong L, Yang L. Oral multivalent epitope vaccine, based on UreB, HpaA, CAT, and LTB, for prevention and treatment of Helicobacter pylori infection in C57BL / 6 mice. Helicobacter 2021; 26:e12807 [View Article] [PubMed]
     [Google Scholar]
  59. Court M, Robinson PA, Dixon MF, Jeremy AHT, Crabtree JE. The effect of gender on Helicobacter felis-mediated gastritis, epithelial cell proliferation, and apoptosis in the mouse model. J Pathol 2003; 201:303–311 [View Article] [PubMed]
     [Google Scholar]
  60. Thung I, Aramin H, Vavinskaya V, Gupta S, Park JY et al. Review article: the global emergence of Helicobacter pylori antibiotic resistance. Aliment Pharmacol Ther 2016; 43:514–533 [View Article] [PubMed]
     [Google Scholar]
  61. WHO Prioritization of pathogens to guide discovery, research and development of new antibiotics for drug-resistant bacterial infections, including tuberculosis. Geneva 2017
     [Google Scholar]
  62. Zhou Z, Dong H, Huang Y, Yao S, Liang B et al. Recombinant Bacillus subtilis spores expressing cholera toxin B subunit and Helicobacter pylori urease B confer protection against H. pylori in mice. J Med Microbiol 2017; 66:83–89 [View Article] [PubMed]
     [Google Scholar]
  63. Day AS, Jones NL, Policova Z, Jennings HA, Yau EK et al. Characterization of virulence factors of mouse-adapted Helicobacter pylori strain SS1 and effects on gastric hydrophobicity. Dig Dis Sci 2001; 46:1943–1951 [View Article] [PubMed]
     [Google Scholar]
  64. Ferrero RL, Avé P, Ndiaye D, Bambou J-C, Huerre MR et al. NF-kappaB activation during acute Helicobacter pylori infection in mice. Infect Immun 2008; 76:551–561 [View Article] [PubMed]
     [Google Scholar]
  65. Lee A, Fox JG, Otto G, Murphy J. A small animal model of human Helicobacter pylori active chronic gastritis. Gastroenterology 1990; 99:1315–1323 [View Article] [PubMed]
     [Google Scholar]
  66. Su H, Tissera K, Jang S, Choi YH, Kim A et al. Evolutionary mechanism leading to the multi-cagA genotype in Helicobacter pylori. Sci Rep 2019; 9:1–3 [View Article] [PubMed]
     [Google Scholar]
  67. Baldwin DN, Shepherd B, Kraemer P, Hall MK, Sycuro LK et al. Identification of Helicobacter pylori genes that contribute to stomach colonization. Infect Immun 2007; 75:1005–1016 [View Article] [PubMed]
     [Google Scholar]
  68. Lowenthal AC, Hill M, Sycuro LK, Mehmood K, Salama NR et al. Functional analysis of the Helicobacter pylori flagellar switch proteins. J Bacteriol 2009; 191:7147–7156 [View Article] [PubMed]
     [Google Scholar]
  69. Oertli M, Noben M, Engler DB, Semper RP, Reuter S et al. Helicobacter pylori γ-glutamyl transpeptidase and vacuolating cytotoxin promote gastric persistence and immune tolerance. Proc Natl Acad Sci U S A 2013; 110:3047–3052 [View Article] [PubMed]
     [Google Scholar]
  70. Marchetti M, Aricò B, Burroni D, Figura N, Rappuoli R et al. Development of a mouse model of Helicobacter pylori infection that mimics human disease. Science 1995; 267:1655–1658 [View Article] [PubMed]
     [Google Scholar]
  71. Kundu P, Mukhopadhyay AK, Patra R, Banerjee A, Berg DE et al. Cag pathogenicity island-independent up-regulation of matrix metalloproteinases-9 and -2 secretion and expression in mice by Helicobacter pylori infection. J Biol Chem 2006; 281:34651–34662 [View Article] [PubMed]
     [Google Scholar]
  72. Dailidiene D, Dailide G, Ogura K, Zhang M, Mukhopadhyay AK et al. Helicobacter acinonychis: genetic and rodent infection studies of a Helicobacter pylori-like gastric pathogen of cheetahs and other big cats. J Bacteriol 2004; 186:356–365 [View Article] [PubMed]
     [Google Scholar]
  73. Ermak TH, Giannasca PJ, Nichols R, Myers GA, Nedrud J et al. Immunization of mice with urease vaccine affords protection against Helicobacter pylori infection in the absence of antibodies and is mediated by MHC class II-restricted responses. J Exp Med 1998; 188:2277–2288 [View Article] [PubMed]
     [Google Scholar]
  74. Kleanthous H, Myers GA, Georgakopoulos KM, Tibbitts TJ, Ingrassia JW et al. Rectal and intranasal immunizations with recombinant urease induce distinct local and serum immune responses in mice and protect against Helicobacter pylori infection. Infect Immun 1998; 66:2879–2886 [View Article] [PubMed]
     [Google Scholar]
  75. Mégraud F, Lehours P, Vale FF. The history of Helicobacter pylori: from phylogeography to paleomicrobiology. Clin Microbiol Infect 2016; 22:922–927 [View Article] [PubMed]
     [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jmm/10.1099/jmm.0.001786
Loading
Genomic and vaccine preclinical studies reveal a novel mouse-adapted Helicobacter pylori model for the hpEastAsia genotype in Southeast Asia
J. Med. Microbiol. 73, 001786 (2024); https://doi.org/10.1099/jmm.0.001786
/content/journal/jmm/10.1099/jmm.0.001786
/content/journal/jmm/10.1099/jmm.0.001786
Loading

Data & Media loading...

Supplements

Supplementary material 1

PDF

Most cited Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error