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Journal of Clinical Microbiology, August 2009, p. 2426-2434, Vol. 47, No. 8
0095-1137/09/$08.00+0     doi:10.1128/JCM.00159-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

CagA and VacA Polymorphisms Do Not Correlate with Severity of Histopathological Lesions in Helicobacter pylori-Infected Greek Children{triangledown} ,{dagger}

Dionyssios N. Sgouras,1* Effrosini G. Panayotopoulou,1 Konstantinos Papadakos,1 Beatriz Martinez-Gonzalez,1 Aikaterini Roumbani,2 Joanna Panayiotou,2 Cathy vanVliet-Constantinidou,2 Andreas F. Mentis,1 and Eleftheria Roma-Giannikou2

Laboratory of Medical Microbiology, Hellenic Pasteur Institute, Athens, Greece,1 First Department of Pediatrics, Athens University School of Medicine, Athens, Greece2

Received 26 January 2009/ Returned for modification 18 March 2009/ Accepted 11 June 2009

The presence of various numbers of EPIYA tyrosine phosphorylation motifs in the CagA protein of Helicobacter pylori has been suggested to contribute to pathogenesis in adults. In this prospective study, we characterized H. pylori isolates from symptomatic children, with reference to the diversity of functional EPIYA motifs in the CagA protein and vacA isotypes, and assessed the potential correlation with the histopathological manifestations of the infection. We analyzed 105 H. pylori isolates from 98 children and determined the diversity of EPIYA motifs in CagA by amplification and sequencing of the 3' variable region of the cagA gene as well as vacA isotypes for the signal, middle, and intermediate regions. CagA phosphorylation and levels of secreted IL-8 were determined following in vitro infection of AGS gastric epithelial cells. Histopathological evaluation of H. pylori colonization, activity, and severity of the associated gastritis was performed according to the updated Sydney criteria. EPIYA A (GLKN[ST]EPIYAKVNKKK), EPIYA B (Q[V/A]ASPEPIY[A/T]QVAKKVNAKI), and EPIYA C (RS[V/A]SPEPIYATIDDLG) motifs were detected in the ABC (46.6%) and ABCC (17.1%) combinations. No isolates harboring more than two EPIYA C motifs in CagA were found. The presence of isogenic strains with variable numbers of CagA EPIYA C motifs within the same patient was detected in seven cases. Occurrence of increasing numbers of EPIYA C motifs correlated strongly with presence of a high-vacuolation (s1 or s2/i1/m1) phenotype and age. A weak positive correlation was observed between vacuolating vacA genotypes and presence of nodular gastritis. However, CagA- and VacA-dependent pathogenicities were not found to contribute to severity of histopathology manifestations in H. pylori-infected children.

* Corresponding author. Mailing address: Laboratory of Medical Microbiology, Hellenic Pasteur Institute, 127 Vas. Sofias Avenue, 115 21 Athens, Greece. Phone: 30210-6478824. Fax: 30210-6440171. E-mail: sgouras{at}pasteur.gr

{triangledown} Published ahead of print on 17 June 2009.

{dagger} Supplemental material for this article may be found at http://jcm.asm.org/.

Journal of Clinical Microbiology, August 2009, p. 2426-2434, Vol. 47, No. 8
0095-1137/09/$08.00+0     doi:10.1128/JCM.00159-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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