The role of Helicobacter pylori outer membrane proteins in adherence and pathogenesis

Helicobacter pylori is one of the most successful human pathogens, whichcolonizes the mucus layer of the gastric epithelium of more than 50% of the world’spopulation. This curved, microaerophilic, Gram-negative bacterium induces a chronicactive gastritis, often asymptomatic, in all infected individuals. In some cases, this gastritisevolves to more severe diseases such as peptic ulcer disease, gastric adenocarcinoma, andgastric mucosa-associated lymphoid tissue lymphoma. H. pylori has developed a unique setof factors, actively supporting its successful survival and persistence in its natural hostileecological niche, the human stomach, throughout the individual’s life, unless treated. In thehuman stomach, the vast majority of H. pylori cells are motile in the mucus layer lining,but a small percentage adheres to the epithelial cell surfaces. Adherence to the gastricepithelium is important for the ability of H. pylori to cause disease because this intimateattachment facilitates: (1) colonization and persistence, by preventing the bacteria frombeing eliminated from the stomach, by mucus turnover and gastric peristalsis; (2) evasionfrom the human immune system and (3) efficient delivery of proteins into the gastric cell,such as the CagA oncoprotein. Therefore, bacteria with better adherence propertiescolonize the host at higher densities. H. pylori is one of the most genetically diversebacterial species known and is equipped with an extraordinarily large set of outermembrane proteins, whose role in the infection and persistence process will be discussed in this review, as well as the different receptor structures that have been so far described for mucosal adherence

Helicobacter pylori—Associated Dyspepsia in Paediatrics

Helicobacter pylori ubiquitously infects the human gastric mucosa since time immemorial,predictably before the man’s diaspora out of East Africa around 58,000 years ago [1].Colonization may have been somehow beneficial for human carriers, allowing the coevolutionof this gram-negative bacterium and its host over the centuries. Yet, at leastnowadays [2], this may not be a peaceful association, with infection almost invariablycausing an acute host immune response. However, in a fully adapted manner, H. pyloriavoids recognition and, thus, clearance, by the host immune system, with both infectionand the consequent gastritis persisting throughout the patients’ life. The clinical outcomeof this persistence is dependent on a sophisticated crosstalk between the host and thepathogen. If often asymptomatic, the H. pylori-associated non-ulcer dyspepsia is clearly thestrongest aetiological factor for severe gastric diseases that will develop late in adult life ina minority of infected patients, i.e., peptic ulcer disease, both gastric and duodenal ulcers,and gastric cancer, namely, adenocarcinoma and mucosa associated lymphoid tissue(MALT) lymphoma (reviewed in [3]). Peptic ulcer disease rarely occurs soon after H. pyloriinfection [4-8] that generally starts in childhood; this presumably reflects marked differencesin the virulence [9-16] and/or in the susceptibility of young patients [17-19].This chapter, focussing on the paediatric population, seeks to explore: the prevalence of H.pylori infection; the molecular mechanism used by H. pylori during colonization and infection;the role of this bacterium in the development of peptic ulcer-related organic dyspepsia; andthe genetic/proteome profile of the H. pylori-strains associated with peptic ulcer disease.BNP Paribas, Sociedade Portuguesa de Gastrenterologi

Positive selection in the evolution of Helicobacter pylori outer membrane proteins

Homologous recombination in Helicobacter pylori has been extensively described to occur via Outer Membrane Proteins (OMPs), regulating protein expression and generating allelic diversity, while the importance of single nucleotide polymorphisms (SNP) remains little studied. We used an OMP-encoding gene, homC, as a model to evaluate the weight of positive selection in the evolution of H. pylori, by using G200 sequences obtained from strains collected worldwide. N-site and branch-site phylogenetic analysis by maximum likelihood models were used to identify specific codons that may be important in homC evolution, and to evaluate the impact of selective pressure on the geographic segregation of strains, respectively. The N-site overall analysis showed that 14 of the 742 (1.9%) homC codons are likely under positive selection (likelihood-ratio test (LRT), p < 10-61). Four of these codons are located in the most variable allelic gene middle region, probably reflecting recombination-derived hitchhiking events. On the other hand, eight codons are located in the more conserved 5¢and 3¢ gene regions, although the significance of this distribution remains to be clarified. Branch-site analysis revealed 36 codons (4.9%) under positive selection (LRT, p < 10-41), showing a non-random distribution, and 89% of these particular codons (p < 10-3) support the phylogenetic segregation of European strains from both African and East Asian strains. The lack of visible recombination within this segment suggests an important biological role of point mutations in the evolution of H. pylori OMPs. In conclusion, homC SNP analysis suggests that, besides recombination, positive selection contributes as well to the evolution of H. pylori OMPs

Clostridium difficile: diversidade genética e perfis de suscetibilidade aos antimicrobianos

Este trabalho teve como objetivo estudar a variabilidade genética e o perfil de suscetibilidade aos antimicrobianos de estirpes de C. difficile recebidas no Instituto Nacional de Saúde Doutor Ricardo Jorge (INSA) entre julho de 2012 e dezembro de 2014

Overview of Clostridium difficile Infection: Life Cycle, Epidemiology, Antimicrobial Resistance and Treatment

The use of antimicrobial agents and acquired resistances explains in part the emergence and spreading of epidemic strains of Clostridium difficile. Continued use of antimicrobial therapy still represents an acute danger in triggering the emergence and spreading of new resistant and multiresistant strains including against first-line antibiotics. We examine the pathway of peptidoglycan synthesis in this organism and associated resistances, as well as resistance to other classes of antibiotics. The life cycle of C. difficile involves growth, spore formation and germination. Spores endow the organism with a formidable capacity of persistence in the environment and in the host, resistance, dissemination and infectious potential. Highly resistant spores produced by antibiotic-resistant/multiresistant strains may be one of the most serious challenges we face in what concerns the containment of C. difficile. Finally, we review recent developments in the treatment and prevention of C. difficile infection

High worldwide conservation of a Helicobacter pylori outer membrane protein

The genetic diversity and evolution of homD, coding for Helicobacter pylori outer membrane protein (OMP) was investigated in a panel of approximately 200 clinical and reference strains, isolated from patients from different geographical origins and presenting different gastric diseases. PCR, sequencing and bioinformatics analyses were used. The homD gene was present in all strains, at a conserved locus, and showed a low genomic diversity, displaying high similarity at both nucleotide and amino acid level. A similarity plot analysis also showed a high level of sequence conservation, although a small region (~30 nucleotides) differed between Western strains and the other strains (East Asian/Ameridian and African). This region was also found in some allelic variants of another hom family member, the homC gene, suggesting the existence of recombination events between these two OMP encoding genes. Sequence analysis of the HomD predicted protein showed a N terminus region with a variable number of KP motif repeats (2 9 KP), with a correlation between the lowest number of KP motif repeats (£4 KP) and peptic ulcer disease and the highest number of repeats (£7 KP) and gastritis. In silico analysis of the HomD protein showed that the region of KP motif repeats exhibits a strong hydrophilicity and antigenicity and a high probability of being exposed to the bacterial surface, suggesting that HomD is immunogenic. These results suggest that homD gene is an important H. pylori antigen and, because of its high global conservation, it is likely to constitute a new vaccine target

Diversity and phylogeny of the Helicobacter pylori outer membrane protein-encoding gene homC

The genetic diversity and evolution of the homC gene was evaluated in a panel of approximately 200 clinical and reference strains, isolated from patients from different geographical origins and presenting different gastric diseases. PCR, sequencing and bioinformatics analyses were used. All the strains tested harboured a complete homC gene at a conserved locus. Phylogenetic reconstruction of homC showed a geographical segregation, with three predominant groups: Western, East Asian/Amerindian and African. A similarity plot analysis suggested a conserved profile of gene segmentation, where three segments were defined. In the first segment (5¢ end extremity), sequences were separated according to the geographical origin of the strain. A higher level of diversity (<50%) was observed in the middle segment, while the third segment (3¢ end extremity) was the most conserved (~90%). In the middle segment, eight allelic variants were identified, with geographic specificity regarding the most prevalent ones. The AI allele was predominant and exclusive of Western strains. The AII allele was predominant in African strains and was the only allele present in the three geographical groups. The AIV allele was predominant in East Asian/ Amerindian strains and was not observed in Western strains. The Western group showed greater molecular distance while the sequences from the East Asian/ Amerindian group were the closest. Overall, the regular presence of homC and its allelic variability suggest that this gene is a good candidate to be part of the pool of H. pylori outer membrane proteins involved in bacterial persistence