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Authors: Nell, Sandra and Estibariz, Iratxe and Krebes, Juliane and Bunk, Boyke and Graham, David Y and Overmann, Jörg and Song, Yi and Spröer, Cathrin and Yang, Ines and Wex, Thomas and Korlach, Jonas and Malfertheiner, Peter and Suerbaum, Sebastian

Helicobacter pylori is remarkable for its genetic variation. Yet little isknown about its genetic changes during early stages of human infection, as the bacteria adapt to their new environment. We analyzed genome and methylome variations in a fully virulent strain of H pylori strain during experimental infection.We performed a randomized Phase 1 and 2, observer-blind, placebo-controlled, study of 12 healthy, H pylori-negative adults in Germany from October 2008 through March 2010. The volunteers were given a prophylactic vaccine candidate (n=7) or placebo (n=5) and then challenged with H pylori strain BCM-300. Biopsy samples were collected and H pylori were isolated. Genomes of the challenge strain and 12 re-isolates, obtained 12 weeks after (or in 1 case, 62 weeks after) infection were sequenced by single-molecule, real-time technology, which, in parallel, permitted determination of genome-wide methylation patterns for all strains. Functional effects of genetic changes observed in H pylori strains during human infection were assessed by measuring release of interleukin 8 from AGS cells (to detect cag PAI function), neutral red uptake (to detect vacuolating cytotoxin activity), and adhesion assays.The observed mutation rate was in agreement with rates previously determined from patients with chronic H pylori infections, without evidence of a mutation burst. A loss; of cag PAI function was observed in 3 re-isolates. In addition, 3 re-isolates from the vaccine; group acquired mutations in the vacuolating cytotoxin gene vacA, resulting in loss of; vacuolization activity from gastric epithelial cells. We observed inter-strain variation in; methylomes due to phase variation in genes encoding methyltransferases.We analyzed adaptation of a fully virulent strain of H pylori to 12 differentvolunteers to obtain a robust estimate of the frequency of genetic and epigenetic changes inthe absence of inter-strain recombination. Our findings indicate that the large amount of; genetic variation in H pylori poses a challenge to vaccine development. ClinicalTrials.gov no: NCT00736476. Copyright © 2017 AGA Institute. Published by Elsevier Inc. All rights reserved.

Journal: Gastroenterology
DOI: 10.1053/j.gastro.2017.10.014
Year: 2017

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