Helicobacter pylori vacA genotypes and cagA gene in a series of 383 H. pylori-positive patients

Summary

Background: Only 10-15 % of all patients infected with Helicobacter pylori develop peptic ulcer disease (PUD) or gastric cancer. Apart from immunological factors in the host, virulence determinants of H. pylori such as the vacuolating cytotoxin (VacA) or the cytotoxin-associated protein A (CagA) might represent a predisposition for the development of PUD.

Methods: We studied antral biopsies of 383 H. pylori-positive patients with peptic ulcer disease (PUD) or other H. pylori-related diseases for H. pylori vacA genotypes and the presence of the cagA gene by PCR.

Results: VacA genotypes and cagA status could be completely determined in 357 (93.2 %) of the patients. In 91 (93.8 %) of 97 patients with PUD, the vacA s1 genotype (s1m1, 45; s1m2, 46 patients) was present. The vacA s2m2 genotype was found in only 6 (6.2 %) of 97 patients with PUD. In contrast, 180 (75.3 %) of 239 patients (s1m1, 89; s1m2, 91 patients) without PUD and without gastric malignancies harbored strains with the vacA s1 genotype. The vacA genotype s2m2 was found in 59 (24.7 %) of these patients. The presence of the cagA gene was closely associated with the vacA genotype s1 and found in 124 (88.6 %) and in 113 (80.7 %) of patients with the s1m1 or s1m2 genotypes, respectively, whereas strains with the genotype s2m2 were almost exclusively cagA negative.

Conclusion: Most H. pylori strains found in patients with PUD possess the vacA s1 genotype and the cagA gene. Patients with this type of H. pylori strain but without PUD might be at higher risk of developing PUD. In contrast, the risk for PUD might be significantly decreased in those patients who are infected by H. pylori strains with the vacA s2 genotype lacking the cagA gene.

(Helicobacter-pylori-vacA-Genotypen und cagA-Gen in einer Serie von 383 H. pylori-positiven Patienten)

Hintergrund: Lediglich 10-15 % aller mit Helicobacter pylori infizierten Patienten entwickeln in der Folge ein peptisches Ulkus (PU) oder ein Magenkarzinom. Neben immunologischen Faktoren können Virulenzfaktoren von H. pylori, wie z. B. das vakuolisierende Zytotoxin (VacA) oder das Zytotoxinassozi­ierte Protein A (CagA), bei der Entwicklung eines peptischen Ulkus prädisponierend sein.

Methoden: In Antrumbiopsien von 383 H. pylori-positiven Patienten mit peptischem Ulkus (PU) oder anderen H. pylori-assoziierten Erkrankungen wurden die H. pylori-vacA-Genotypen und die Präsenz des cagA-Gens mit PCR untersucht.

Ergebnisse: Die H. pylori-vacA-Genotypen und der cagA-Status konnten bei 357 (93,2 %) Patienten vollständig bestimmt werden. Bei 91 (93,8 %) von 97 Patienten mit einem PU lagen H. pylori-Stämme mit dem vacA-Genotyp s1 (s1m1: 45; s1m2: 46 Patienten) vor. Der vacA-Genotyp s2m2 war nur bei 6 (6,2 %) von 97 Patienten mit PU vorhanden. Dagegen hatten 180 (75,3 %) von 239 Patienten ohne PU und ohne ein Magenmalignom Stämme mit dem vacA-Genotyp s1 (s1m1: 89; s1m2: 91 Patienten). Der vacA-s2m2-Genotyp wurde bei 59 (24,7 %) dieser Patienten nachgewiesen. Der Nachweis des cagA-Gens war eng mit dem vacA-s1-Genotyp assoziiert und wurde bei 124 (88,6 %) bzw. 113 (80,7 %) der Patienten mit den vacA-Genotypen s1m1 oder s1m2 gefunden, während Stämme mit dem vacA-Genotyp s2m2 fast ausschließlich cagA-negativ waren.

Schlussfolgerung: Bei nahezu allen H. pylori-positiven Pa­tienten mit peptischem Ulkus liegen H. pylori-Stämme mit dem vacA-Genotyp s1 und mit cagA-Gen vor. Patienten ohne PU mit H. pylori-Stämmen dieses Genotyps haben vermutlich ein höheres Risiko, ein peptisches Ulkus zu entwickeln. Dagegen erscheint das Risiko für ein Ulkus bei Patienten mit H. pylori-Stämmen des vacA-Genotyps s2 und ohne cagA-Gen signifikant verringert zu sein.

Key words

Helicobacter Pylori - Gastric Biopsies - Polymerase Chain Reaction - Peptic Ulcer Disease

Schlüsselwörter

Helicobacter-pylori - Magenbiopsien - Polymerasekettenreaktion - peptisches Ulkus

References

• 1 Warren J R, Marshall B J. Unidentified curved bacilli on gastric ­epithelium in active chronic gastritis.  Lancet. 1983;  I 1273-1274
• 2 Hentschel I, Brandstätter G, Dragosics B. et al . Effect of ranitidine and amoxicillin plus metronidazole on the eradication of Helicobacter pylori in the pathogenesis of duodenal and gastric ulcer.  N Engl J Med. 1993;  328 308-312
• 3 Parsonnet J, Friedman G D, Vandersteen D P. et al . Helicobacter pylori infection and the risk of gastric carcinoma.  N Engl J Med. 1991;  325 1127-1131
• 4 Wotherspoon A C, Ortiz-Hidalgo C, Falzon M R. et al . Helicobacter-pylori-associated gastritis and primary B-cell gastric lymphoma.  Lancet. 1991;  338 1175-1176
• 5 Stolte M. Helicobacter pylori gastritis and gastric MALT-lymphoma.  Lancet. 1992;  339 745-746
• 6 Leunk R D, Johnson P T, David B C. et al . Cytotoxic activity in broth-culture filtrates of Campylobacter pylori .  J Med Microbiol. 1998;  26 93-99
• 7 Figura N, Guglielmetti P, Rossolini A. et al . Cytotoxin production by Campylobacter pylori strains isolated from patients with ­peptic ulcers and from patients with chronic gastritis.  J Clin ­Microbiol. 1989;  27 225-226
• 8 Cover T L, Dooley C P, Blaser M J. Characterisation of and human serologic response to proteins in Helicobacter pylori broth culture supernatants with vacuolizing cytotoxin activity.  Invect Immun. 1990;  58 603-610
• 9 Crabtree J E, Taylor J D, Wyatt J I. et al . Mucosal IgA recognition of Helicobacter pylori 120 kDA protein, peptic ulceration and gastric pathology.  Lancet. 1991;  59 1264-1270
• 10 Covacci A, Censini S, Bugnoli M. et al . Molecular characterization of the 128-kDa immunodominant antigen of Helicobacter pylori associated with cytotoxicity and duodenal ulcer.  Proc Natl Acad Sci USA. 1993;  9 5791-5795
• 11 Cover T L, Cao P, Lind C D. et al . Correlation between vacuolating cytotoxin production by Helicobacter pylori isolates in vitro and in vivo.  Infect Immun. 1993;  61 5008-5012
• 12 Cover T L, Tummuru M KR, Cao P. et al . Divergence of genetic sequences for the vacuolating cytotoxin among Helicobacter pylori strains.  J Biol Chem. 1994;  269 10566-10573
• 13 Schmitt W, Haas R. Genetic analysis of the Helicobacter pylori ­vacuolating cytotoxin: Structural similarities with the IgA protease type of exported protein.  Mol Microbiol. 1994;  12 307-319
• 14 Goosens H, Glupczynski Y, Burette A. et al . Role of the vacuolating toxin from Helicobacter pylori in the pathogenesis of duodenal and gastric ulcer.  Med Microbiol Lett. 1992;  1 153-159
• 15 Atherton J C, Cao P, Peek R M. et al . Mosaicism in vacuolating ­cytotoxin alleles of Helicobacter pylori .  J Biol Chem. 1995;  27 17771-17777
• 16 Atherton J C, Peek R M, Tham K T. et al . Clinical and pathological importance of heterogeneity in vacA, the vacuolating cytotoxin gene of Helicobacter pylori .  Gastroenterology. 1997;  112 92-99
• 17 Rudi J, Kolb C, Maiwald M. et al . Diversity of Helicobacter pylori vacA and cagA genes and relationship to VacA and CagA protein expression, cytotoxin production, and associated diseases.  J Clin Microbiol. 1998;  36 944-948
• 18 Van Dorn L J, Figueiredo C, Sanna R. et al . Clinical relevance of the cagA, vacA, and iceA status of Helicobacter pylori .  Gastroenterology. 1998;  115 58-66
• 19 Censini S, Lange C, Xiang Z. et al . Cag, a pathogenicity island of Helicobacter pylori, encodes type I-specific and disease-associated virulence factors.  Proc Natl Acad Sci USA. 1996;  93 14648-14653
• 20 Tummuru M KR, Sharma S A, Blaser M J. Helicobacter pylori picB, a homologue of the Bordetella pertussis toxin secretion protein, is required for induction of IL-8 in gastric epithelial cells.  Mol Microbiol. 1995;  18 867-876
• 21 Tummuru M KR, Cover T L, Blaser M J. Cloning and expression of a high-molecular-mass major antigen of Helicobacter pylori: Evidence of linkage to cytotoxin production.  Infect Immun. 1993;  61 1799-1809
• 22 Crabtree J E, Figura N, Taylor J D. et al . Expression of 120 kilodalton protein and cytotoxicity in Helicobacter pylori .  J Clin Pathol. 1992;  45 733-734
• 23 Xiang Z, Censini S, Bayelli P F. et al . Analysis of expression of CagA and VacA virulence factors in 43 strains of Helicobacter pylori ­reveals that clinical isolates can be divided into two major types and that CagA is not necessary for expression of the vacuolating cytotoxin.  Infect Immun. 1995;  63 94-98
• 24 Rudi J, Kolb C, Maiwald M. et al . Serum antibodies against Helicobacter pylori proteins CagA and VacA are associated with increased risk for gastric adenocarcinoma.  Dig Dis Sci. 1997;  42 1652-1659
• 25 Warburton V J, Everett S, Mapstone N P. et al . Clinical and histological associations of cagA and vacA genotypes in Helicobacter pylori .  J Clin Pathol. 1998;  51 55-61
• 26 Jenks P J, Megraud F, Labigne A. Clinical outcome after infection with Helicobacter pylori does not appear to be reliably predicted by the presence of any of the genes of the pathogenicity island.  Gut. 1998;  43 752-758
• 27 Van Doorn L J, Figueiredo C, Megraud F. et al . Geographic distribution of vacA allelic types of Helicobacter pylori .  Gastroenterology. 1999;  116 823-830
• 28 Yamaoka Y, Kodama T, Gutierrez O. et al . Relationship between Helicobacter pylori iceA, cagA, and vacA status and clinical outcome: Studies in four different countries.  J Clin Microbiol. 1999;  37 2274-2279
• 29 Schultze V, Hackelsberger A, Günther T. et al . Differing patterns of Helicobacter pylori gastritis in patients with duodenal, prepyloric, and gastric ulcer disease.  Scand J Gastroenterol. 1998;  33 137-142
• 30 Hamlet A, Thoreson A C, Nilsson O. et al . Duodenal Helicobacter pylori infection differs in cagA genotype between asymptomatic subjects and patients with duodenal ulcers.  Gastroenterology. 1999;  116 259-268
• 31 Rudi J, Kuck D, Strand S. et al . Involvement of the CD95 (APO-1/Fas) receptor and ligand system in Helicobacter pylori-induced gastric epithelial apoptosis.  J Clin Invest. 1998;  102 1506-1514
• 32 Blum A L, Talley N J, OMorain C. et al . Lack of effect of treating Helicobacter pylori infection in patients with nonulcer dyspepsia.  N Engl J Med. 1998;  339 1875-1881
• 33 McColl K, Murray L, El-Omar E. et al . Symptomatic benefit from eradicating Helicobacter pylori infection in patients with nonulcer dyspepsia.  N Engl J Med. 1998;  339 1869-1874
• 34 Hoshina S, Kahn S M, Jiang W. et al . Direct detection and amplification of Helicobacter pylori ribosomal 16S gene segments from gastric endoscopic biopsies.  Diagn Microbiol Infect Dis. 1990;  13 473-479
• 35 Weiss J, Mecca J, da Silva E. et al . Comparison of PCR and other diagnostic techniques for detection of Helicobacter pylori infection in dyspeptic patients.  J Clin Microbiol. 1994;  32 1663-1668
• 36 Rollan A, Giancaspero R, Arrese M. et al . Accuracy of invasive and noninvasive tests to diagnose Helicobacter pylori infection after antibiotic treatment.  Am J Gastroenterol. 1997;  92 1268-1274
• 37 Rudi J, Rudy A, Maiwald M. et al . Direct determination of Helicobacter pylori vacA genotypes and cagA gene in gastric biopsies and relationship to gastrointestinal diseases.  Am J Gastroenterol. 1999;  94 1525-1531
• 38 Maiwald M, Kissel K, Srimuang S. et al . Comparison of polymerase chain reaction and conventional culture for the detection of legionellas in hospital water.  J Appl Bacteriol. 1994;  76 216-225
• 39 Riley L K, Franklin C L, Hook R R. et al . Identification of murine Helicobacters by PCR and restriction enzyme analyses.  J Clin ­Microbiol. 1996;  34 942-946
• 40 Maiwald M, Ditton H -J, Sonntag H -G. et al . Characterization of contaminating DNA in Taq polymerase which occurs during amplification with a primer set for Legionella 5S ribosomal RNA.  Mol Cell Prob. 1994;  8 11-14
• 41 Han S R, Schreiber H J, Bhakdi S. et al . VacA genotypes and genetic diversity in clinical isolates of Helicobacter pylori .  Clin Diagn Lab Immunol. 1998;  5 139-145
• 42 Strobel S, Bereswill S, Balig P. et al . Identification and analysis of a new vacA genotype variant of Helicobacter pylori in different patient groups in Germany.  J Clin Microbiol. 1998;  36 1285-1289
• 43 Stephens J C, Stewart J A, Folwell A M. et al . Helicobacter pylori cagA status, vacA genotypes and ulcer disease.  Eur J Gastroenterol Hepatol. 1998;  1 381-384
• 44 The European Helicobacter pylori Study Group   (EHPSG). Current European concepts in the management of Helicobacter pylori infection. The Maastricht Consensus Report.  Gut. 1997;  41 8-13
• 45 Van Doorn L J, Figueiredo C, Rossau R. et al . Typing of Helicobacter pylori vacA gene and detection of cagA gene by PCR and reverse hybridization.  J Clin Microbiol. 1998;  36 1271-1275
• 46 Fibura N, Vindigni C, Covacci A. et al . CagA positive and negative Helicobacter pylori strains are simultaneously present in the stomach of most patients with non-ulcer dyspepsia: Relevance of histological damage.  Gut. 1998;  42 772-778

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