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Ulcerogenic Helicobacter pylori strains isolated from children: a contribution to get insight into the virulence of the bacteria.

Vitoriano I, Saraiva-Pava KD, Rocha-Gonçalves A, Santos A, Lopes AI, Oleastro M, Roxo-Rosa M - PLoS ONE (2011)

Bottom Line: Corroborating this, our in vitro infection assays comparing a pool of five H. pylori strains isolated from children with PUD to a pool of five other pediatric clinical isolates associated with non-ulcer dyspepsia (NUD) showed the greater ability of PUD strains to induce a marked decrease in the viability of gastric cells and to cause severe damage in the cells cytoskeleton as well as an impairment in the production/secretion of mucins.To uncover virulence features, we compared the proteome of these two groups of H. pylori strains.In conclusion, the enhanced virulence of the pediatric ulcerogenic H. pylori strains may result from a synergy between their natural ability to better adapt to the hostile human stomach and the expression of the established virulence factors.

View Article: PubMed Central - PubMed

Affiliation: Faculdade de Engenharia, Universidade Católica Portuguesa, Rio de Mouro, Portugal.

ABSTRACT
Infection with Helicobacter pylori is the major cause for the development of peptic ulcer disease (PUD). In children, with no other etiology for the disease, this rare event occurs shortly after infection. In these young patients, habits of smoking, diet, consumption of alcohol and non-steroid anti-inflammatory drugs and stress, in addition to the genetic susceptibility of the patient, represent a minor influence. Accordingly, the virulence of the implicated H. pylori strain should play a crucial role in the development of PUD. Corroborating this, our in vitro infection assays comparing a pool of five H. pylori strains isolated from children with PUD to a pool of five other pediatric clinical isolates associated with non-ulcer dyspepsia (NUD) showed the greater ability of PUD strains to induce a marked decrease in the viability of gastric cells and to cause severe damage in the cells cytoskeleton as well as an impairment in the production/secretion of mucins. To uncover virulence features, we compared the proteome of these two groups of H. pylori strains. Two-dimensional gel electrophoresis followed by mass-spectrometry allowed us to detect 27 differentially expressed proteins between them. In addition to the presence of genes encoding well established virulence factors, namely cagA, vacAs1, oipA "on" status, homB and jhp562 genes, the pediatric ulcerogenic strains shared a proteome profile characterized by changes in the abundance of: motility-associated proteins, accounting for higher motility; antioxidant proteins, which may confer increased resistance to inflammation; and enzymes involved in key steps in the metabolism of glucose, amino acids and urea, which may be advantageous to face fluctuations of nutrients. In conclusion, the enhanced virulence of the pediatric ulcerogenic H. pylori strains may result from a synergy between their natural ability to better adapt to the hostile human stomach and the expression of the established virulence factors.

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Related in: MedlinePlus

Profile of the soluble proteome of the 10 studied pediatric H. pylori strains.These include five strains associated with NUD (173/00, 207/99, 228/99, 655/99 and 1786/05), four from patients affected by DU (1089/03, 1152/04, 1198/04 and 1846/05) and a strain associated with GU (499/02). Proteins from the total biomass recovered from a 24 h grown plate were separated by 2DE, i.e., first in a non-linear pH 3–11 and than in a 7–16% (w/v) SDS-PAGE. After CBB-staining, gels were analyzed using ImageMaster™ 2-D Platinum software. Black arrow indicates spots that were more abundant in PUD (DU+GU) strains when compared to NUD strains; Dotted arrow indicates spots that were less abundant in PUD (DU+GU) strains when compared to NUD strains; Dashed arrow indicates the spot that was shifted right in DU strains when compared to NUD strains. Spot numbers indicated in the 2DE maps are the same as used in Table 2.
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pone-0026265-g003: Profile of the soluble proteome of the 10 studied pediatric H. pylori strains.These include five strains associated with NUD (173/00, 207/99, 228/99, 655/99 and 1786/05), four from patients affected by DU (1089/03, 1152/04, 1198/04 and 1846/05) and a strain associated with GU (499/02). Proteins from the total biomass recovered from a 24 h grown plate were separated by 2DE, i.e., first in a non-linear pH 3–11 and than in a 7–16% (w/v) SDS-PAGE. After CBB-staining, gels were analyzed using ImageMaster™ 2-D Platinum software. Black arrow indicates spots that were more abundant in PUD (DU+GU) strains when compared to NUD strains; Dotted arrow indicates spots that were less abundant in PUD (DU+GU) strains when compared to NUD strains; Dashed arrow indicates the spot that was shifted right in DU strains when compared to NUD strains. Spot numbers indicated in the 2DE maps are the same as used in Table 2.

Mentions: Total soluble protein extracts from each of the 10 H. pylori strains were resolved by 2-dimensional gel electrophoresis (2DE), first in a non-linear pH range of 3–11 and then in a 7–16% (w/v) SDS-PAGE. After Coomassie Brilliant Blue (CBB) staining, the digitalized images of the gels were analyzed using the ImageMaster™ 2D Platinum software. Protein spots were automatically detected and manually corrected and the best resolved 2DE-gel, that of NUD-associated H. pylori strain 655/99, was used as the reference map during computer assisted analysis. After automatic matching, followed by a manual correction, gels were separated into two classes, NUD and PUD, according to the pathology with which the strain was associated. Gel-to-gel matching was confirmed by MS identification of matched spots from different 2DE gels. The 2DE maps obtained for each group of strains were found to be highly reproducible and consistent (Figure 3). No statistically difference was observed for the number of detected spots (on average 485±77 vs. 443±43 in NUD and PUD 2DE gels, respectively) between these two classes.


Ulcerogenic Helicobacter pylori strains isolated from children: a contribution to get insight into the virulence of the bacteria.

Vitoriano I, Saraiva-Pava KD, Rocha-Gonçalves A, Santos A, Lopes AI, Oleastro M, Roxo-Rosa M - PLoS ONE (2011)

Profile of the soluble proteome of the 10 studied pediatric H. pylori strains.These include five strains associated with NUD (173/00, 207/99, 228/99, 655/99 and 1786/05), four from patients affected by DU (1089/03, 1152/04, 1198/04 and 1846/05) and a strain associated with GU (499/02). Proteins from the total biomass recovered from a 24 h grown plate were separated by 2DE, i.e., first in a non-linear pH 3–11 and than in a 7–16% (w/v) SDS-PAGE. After CBB-staining, gels were analyzed using ImageMaster™ 2-D Platinum software. Black arrow indicates spots that were more abundant in PUD (DU+GU) strains when compared to NUD strains; Dotted arrow indicates spots that were less abundant in PUD (DU+GU) strains when compared to NUD strains; Dashed arrow indicates the spot that was shifted right in DU strains when compared to NUD strains. Spot numbers indicated in the 2DE maps are the same as used in Table 2.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3198394&req=5

pone-0026265-g003: Profile of the soluble proteome of the 10 studied pediatric H. pylori strains.These include five strains associated with NUD (173/00, 207/99, 228/99, 655/99 and 1786/05), four from patients affected by DU (1089/03, 1152/04, 1198/04 and 1846/05) and a strain associated with GU (499/02). Proteins from the total biomass recovered from a 24 h grown plate were separated by 2DE, i.e., first in a non-linear pH 3–11 and than in a 7–16% (w/v) SDS-PAGE. After CBB-staining, gels were analyzed using ImageMaster™ 2-D Platinum software. Black arrow indicates spots that were more abundant in PUD (DU+GU) strains when compared to NUD strains; Dotted arrow indicates spots that were less abundant in PUD (DU+GU) strains when compared to NUD strains; Dashed arrow indicates the spot that was shifted right in DU strains when compared to NUD strains. Spot numbers indicated in the 2DE maps are the same as used in Table 2.
Mentions: Total soluble protein extracts from each of the 10 H. pylori strains were resolved by 2-dimensional gel electrophoresis (2DE), first in a non-linear pH range of 3–11 and then in a 7–16% (w/v) SDS-PAGE. After Coomassie Brilliant Blue (CBB) staining, the digitalized images of the gels were analyzed using the ImageMaster™ 2D Platinum software. Protein spots were automatically detected and manually corrected and the best resolved 2DE-gel, that of NUD-associated H. pylori strain 655/99, was used as the reference map during computer assisted analysis. After automatic matching, followed by a manual correction, gels were separated into two classes, NUD and PUD, according to the pathology with which the strain was associated. Gel-to-gel matching was confirmed by MS identification of matched spots from different 2DE gels. The 2DE maps obtained for each group of strains were found to be highly reproducible and consistent (Figure 3). No statistically difference was observed for the number of detected spots (on average 485±77 vs. 443±43 in NUD and PUD 2DE gels, respectively) between these two classes.

Bottom Line: Corroborating this, our in vitro infection assays comparing a pool of five H. pylori strains isolated from children with PUD to a pool of five other pediatric clinical isolates associated with non-ulcer dyspepsia (NUD) showed the greater ability of PUD strains to induce a marked decrease in the viability of gastric cells and to cause severe damage in the cells cytoskeleton as well as an impairment in the production/secretion of mucins.To uncover virulence features, we compared the proteome of these two groups of H. pylori strains.In conclusion, the enhanced virulence of the pediatric ulcerogenic H. pylori strains may result from a synergy between their natural ability to better adapt to the hostile human stomach and the expression of the established virulence factors.

View Article: PubMed Central - PubMed

Affiliation: Faculdade de Engenharia, Universidade Católica Portuguesa, Rio de Mouro, Portugal.

ABSTRACT
Infection with Helicobacter pylori is the major cause for the development of peptic ulcer disease (PUD). In children, with no other etiology for the disease, this rare event occurs shortly after infection. In these young patients, habits of smoking, diet, consumption of alcohol and non-steroid anti-inflammatory drugs and stress, in addition to the genetic susceptibility of the patient, represent a minor influence. Accordingly, the virulence of the implicated H. pylori strain should play a crucial role in the development of PUD. Corroborating this, our in vitro infection assays comparing a pool of five H. pylori strains isolated from children with PUD to a pool of five other pediatric clinical isolates associated with non-ulcer dyspepsia (NUD) showed the greater ability of PUD strains to induce a marked decrease in the viability of gastric cells and to cause severe damage in the cells cytoskeleton as well as an impairment in the production/secretion of mucins. To uncover virulence features, we compared the proteome of these two groups of H. pylori strains. Two-dimensional gel electrophoresis followed by mass-spectrometry allowed us to detect 27 differentially expressed proteins between them. In addition to the presence of genes encoding well established virulence factors, namely cagA, vacAs1, oipA "on" status, homB and jhp562 genes, the pediatric ulcerogenic strains shared a proteome profile characterized by changes in the abundance of: motility-associated proteins, accounting for higher motility; antioxidant proteins, which may confer increased resistance to inflammation; and enzymes involved in key steps in the metabolism of glucose, amino acids and urea, which may be advantageous to face fluctuations of nutrients. In conclusion, the enhanced virulence of the pediatric ulcerogenic H. pylori strains may result from a synergy between their natural ability to better adapt to the hostile human stomach and the expression of the established virulence factors.

Show MeSH
Related in: MedlinePlus