title: Cholesterol glucosylation by Helicobacter pylori delays internalization and arrests phagosome maturation in macrophages.
abstract: OBJECTIVE
  Helicobacter pylori colonizes the human stomach and contributes to chronic inflammation of the gastric mucosa. H. pylori persistence occurs because of insufficient eradication by phagocytic cells. A key factor of H. pylori, cholesterol-α-glucosyltransferase encoded by capJ that extracts host cholesterol and converts it to cholesteryl glucosides, is important to evade host immunity. Here, we examined whether phagocytic trafficking in macrophages was perturbed by capJ-carrying H. pylori.
  
  
  METHODS
  J774A.1 cells were infected with H. pylori at a multiplicity of infection of 50. Live-cell imaging and confocal microscopic analysis were applied to monitor the phagocytic trafficking events. The viability of H. pylori inside macrophages was determined by using gentamicin colony-forming unit assay. The phagocytic routes were characterized by using trafficking-intervention compounds.
  
  
  RESULTS
  Wild type (WT) H. pylori exhibited more delayed entry into macrophages and also arrested phagosome maturation more than did capJ knockout mutant. Pretreatment of genistein and LY294002 prior to H. pylori infection reduced the internalization of WT but not capJ-knockout H. pylori in macrophages.
  
  
  CONCLUSIONS
  Cholesterol glucosylation by H. pylori interferes with phagosome trafficking via a lipid-raft and PI3K-dependent manner, which retards engulfment of bacteria for prolonged intracellular survival of H. pylori.
authors: Institute of Molecular and Cellular Biology and Department of Life Science, National Tsing Hua University, Hsinchu, Taiwan.
authors: Shin-Yi
authors: SY
authors: Du
authors: Institute of Molecular and Cellular Biology and Department of Life Science, National Tsing Hua University, Hsinchu, Taiwan.
authors: Hung-Jung
authors: HJ
authors: Wang
authors: Institute of Molecular and Cellular Biology and Department of Life Science, National Tsing Hua University, Hsinchu, Taiwan; Biomedical Science and Engineering Center, National Tsing Hua University, Hsinchu, Taiwan.
authors: Hsin-Hung
authors: HH
authors: Cheng
authors: Institute of Molecular and Cellular Biology and Department of Life Science, National Tsing Hua University, Hsinchu, Taiwan.
authors: Sheng-De
authors: SD
authors: Chen
authors: Institute of Molecular and Cellular Biology and Department of Life Science, National Tsing Hua University, Hsinchu, Taiwan.
authors: Lily Hui-Ching
authors: LH
authors: Wang
authors: Institute of Molecular and Cellular Biology and Department of Life Science, National Tsing Hua University, Hsinchu, Taiwan; Biomedical Science and Engineering Center, National Tsing Hua University, Hsinchu, Taiwan. Electronic address: wcwang@mx.nthu.edu.tw.
authors: Wen-Ching
authors: WC
authors: Wang
mesh_terms: D000818:Animals; D002460:Cell Line; D002784:Cholesterol / Q000031:analogs & derivatives* / Q000378:metabolism*; D005964:Glucosyltransferases / Q000235:genetics / Q000378:metabolism*; D016481:Helicobacter Infections / Q000276:immunology / Q000382:microbiology; D016480:Helicobacter pylori / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D057131:Immune Evasion / Q000276:immunology*; D008264:Macrophages / Q000276:immunology*; D021962:Membrane Microdomains / Q000378:metabolism; D051379:Mice; D018613:Microscopy, Confocal; D010587:Phagocytosis / Q000276:immunology; D010588:Phagosomes / Q000276:immunology* / Q000382:microbiology; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism
chemical_list: D002784:Cholesterol; D005964:Glucosyltransferases; D019869:Phosphatidylinositol 3-Kinases; C027578:cholesteryl glucoside; C514935:Hp0421 protein, Helicobacter pylori
keywords: Helicobacter pylori; cholesteryl glucosides; lipid raft; phagocytosis; phagosome maturation; phagosome trafficking