Presence Of Cag Pathogenicity Island Research Articles

Helicobacter pylori Modulates Heptose Metabolite Biosynthesis and Heptose-Dependent Innate Immune Host Cell Activation by Multiple Mechanisms.

Heptose metabolites including ADP-d-glycero-β-d-manno-heptose (ADP-heptose) are involved in bacterial lipopolysaccharide and cell envelope biosynthesis. Recently, heptoses were also identified to have potent proinflammatory activity on human cells as novel microbe-associated molecular patterns. The gastric pathogenic bacterium Helicobacter pylori produces heptose metabolites, which it transports into human cells through its Cag type 4 secretion system. Using H. pylori as a model, we have addressed the question of how proinflammatory ADP-heptose biosynthesis can be regulated by bacteria. We have characterized the interstrain variability and regulation of heptose biosynthesis genes and the modulation of heptose metabolite production by H. pylori, which impact cell-autonomous proinflammatory human cell activation. HldE, a central enzyme of heptose metabolite biosynthesis, showed strong sequence variability between strains and was also variably expressed between strains. Amounts of gene transcripts in the hldE gene cluster displayed intrastrain and interstrain differences, were modulated by host cell contact and the presence of the cag pathogenicity island, and were affected by carbon starvation regulator A (CsrA). We reconstituted four steps of the H. pylori lipopolysaccharide (LPS) heptose biosynthetic pathway in vitro using recombinant purified GmhA, HldE, and GmhB proteins. On the basis of one- and two-dimensional nuclear magnetic resonance (NMR) spectroscopy and mass spectrometry, the structures of major reaction products were identified as β-d-ADP-heptose and β-heptose-1-monophosphate. A proinflammatory heptose-monophosphate variant was also identified for the first time as a novel cell-active product in H. pylori bacteria. Separate purified HldE subdomains and variant HldE allowed us to uncover additional strain variation in generating heptose metabolites. IMPORTANCE Bacterial heptose metabolites, intermediates of lipopolysaccharide (LPS) biosynthesis, are novel microbe-associated molecular patterns (MAMPs) that activate proinflammatory signaling. In the gastric pathogen Helicobacter pylori, heptoses are transferred into host cells by the Cag type IV secretion system, which is also involved in carcinogenesis. Little is known about how H. pylori, which is highly strain variable, regulates heptose biosynthesis and downstream host cell activation. We report here that the regulation of proinflammatory heptose production by H. pylori is strain specific. Heptose gene cluster activity is modulated by the presence of an active cag pathogenicity island (cagPAI), contact with human cells, and the carbon starvation regulator A. Reconstitution with purified biosynthesis enzymes and purified bacterial lysates allowed us to biochemically characterize heptose pathway products, identifying a heptose-monophosphate variant as a novel proinflammatory metabolite. These findings emphasize that the bacteria use heptose biosynthesis to fine-tune inflammation and also highlight opportunities to mine the heptose biosynthesis pathway as a potential therapeutic target against infection, inflammation, and cancer.

Correlation of CagA-Positive Strains of Helicobacter pylori with Topographic Distribution and Chronic gastritis grading

Background: CagA gene is a marker for the presence of Cag pathogenicity island. CagA-positive strains of Helicobacter pylori can identify individuals who have higher risk of developing gastrointestinal diseases. Aim: To discover the correlation of CagA status of Helicobacter pylori with topographic localization of Helicobacter pylori and chronic gastritis grading. Methods: Gastric biopsy specimens were taken from 104 patients. The specimens were obtained from gastric antrum, corpus and incisures for histological and polymerase chain reaction (PCR) studies. The histological chronic gastritis was assessed semi-quantitatively (grades 0-3). The PCR was used for detecting Helicobacter pylori genes and CagA strain. Topographic localization of Helicobacter pylori was classified as gastric antrum and corpus. Results: There were 33 (86.8%) CagA-positive strains of 38 patients with Helicobacter pylori-positive genes. There were no significant differences between topographic localization of Helicobacter pylori - either in the gastric antrum (rho = 0.14, p = 0.40) nor in the corpus (rho = 0.27, p =0.10) and the CagA status of Helicobacter pylori. Conclusion: CagA gene status of Helicobacter pylori does not determine chronic gastritis grading and gastric topographic localization. Keywords: chronic gastritis, cagA gene, Helicobacter pylori, gastric antrum, gastric corpus.