PMID- 12058029
OWN - NLM
STAT- MEDLINE
DCOM- 20021203
LR  - 20210209
IS  - 0021-9258 (Print)
IS  - 0021-9258 (Linking)
VI  - 277
IP  - 37
DP  - 2002 Sep 13
TI  - Microaerophilic conditions permit to mimic in vitro events occurring during in 
      vivo Helicobacter pylori infection and to identify Rho/Ras-associated proteins in 
      cellular signaling.
PG  - 33978-86
AB  - Molecular dissection of the mechanisms underlying Helicobacter pylori infection 
      suffers from the lack of in vitro systems mimicking in vivo observations. A 
      system was developed whereby human epithelial cells (Caco-2) grown as polarized 
      monolayers and bacteria can communicate with each other under culture conditions 
      optimal for each partner. Caco-2 cells grown on filter supports were inserted in 
      a vertical position into diffusion chambers equilibrated with air and 5% CO(2) at 
      their basolateral surface (aerophilic conditions) and 5% CO(2), 5% O(2), 90% N(2) 
      (microaerophilic conditions) in the apical compartment. Remarkably, the 
      epithelial polarized layer was stable under these asymmetric culture conditions 
      for at least 24 h, and the presence of Caco-2 cells was necessary to maintain H. 
      pylori growth. In contrast to previous studies conducted with non-polarized 
      Caco-2 cells and other cell lines kept under aerophilic conditions, we found H. 
      pylori-dependent stimulation of cytokine secretion (MCP-1 (monocyte 
      chemoattractant protein-1), GRO-alpha (growth-regulated oncogene-alpha), RANTES 
      (regulated on activation normal T cell expressed and secreted)). This correlated 
      with nuclear translocation of NF-kappaB p50 and p65 subunits. Tyrosine 
      phosphorylation of nine cellular proteins was induced or enhanced; we identified 
      p120(RasGAP), p190(RhoGAP), p62dok (downstream of tyrosine kinases), and 
      cortactin as H. pylori-inducible targets. Moreover, reduction of H. pylori urease 
      expression was observed in adherent bacteria as compared with bacteria in 
      suspension. In addition to mimicking several observations seen in the inflamed 
      gastric mucosa, the novel in vitro system was allowed to underscore complex 
      cellular events not seen in classical in vitro analyses of microaerophilic 
      bacteria-epithelial cell cross-talk.
FAU - Cottet, Sandra
AU  - Cottet S
AD  - Division of Immunology and Allergy, R & D Laboratory, Centre Hospitalier 
      Universitaire Vaudois, 1005 Lausanne, Switzerland.
FAU - Corthesy-Theulaz, Irene
AU  - Corthesy-Theulaz I
FAU - Spertini, Francois
AU  - Spertini F
FAU - Corthesy, Blaise
AU  - Corthesy B
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20020610
PL  - United States
TA  - J Biol Chem
JT  - The Journal of biological chemistry
JID - 2985121R
RN  - 0 (ARHGAP35 protein, human)
RN  - 0 (ARHGAP5 protein, human)
RN  - 0 (CTTN protein, human)
RN  - 0 (Chemokines)
RN  - 0 (Cortactin)
RN  - 0 (DNA-Binding Proteins)
RN  - 0 (DOK1 protein, human)
RN  - 0 (GAP-associated protein p62)
RN  - 0 (GTPase-Activating Proteins)
RN  - 0 (Guanine Nucleotide Exchange Factors)
RN  - 0 (Microfilament Proteins)
RN  - 0 (NF-kappa B)
RN  - 0 (Nuclear Proteins)
RN  - 0 (Phosphoproteins)
RN  - 0 (RASGRF1 protein, human)
RN  - 0 (RNA-Binding Proteins)
RN  - 0 (Repressor Proteins)
RN  - 0 (p120 GTPase Activating Protein)
RN  - 0 (ras-GRF1)
RN  - EC 3.5.1.5 (Urease)
SB  - IM
MH  - Caco-2 Cells
MH  - Cell Polarity
MH  - Chemokines/metabolism
MH  - Cortactin
MH  - DNA-Binding Proteins/*metabolism
MH  - Epithelial Cells/metabolism
MH  - GTPase-Activating Proteins
MH  - Gastric Mucosa/metabolism
MH  - Guanine Nucleotide Exchange Factors/*metabolism
MH  - Helicobacter Infections/*metabolism
MH  - *Helicobacter pylori/enzymology/growth & development
MH  - Humans
MH  - Microfilament Proteins/*metabolism
MH  - NF-kappa B/metabolism
MH  - Nuclear Proteins/*metabolism
MH  - Phosphoproteins/*metabolism
MH  - Phosphorylation
MH  - RNA-Binding Proteins/*metabolism
MH  - Repressor Proteins
MH  - Urease/biosynthesis
MH  - p120 GTPase Activating Protein/*metabolism
MH  - ras-GRF1
EDAT- 2002/06/12 10:00
MHDA- 2002/12/04 04:00
CRDT- 2002/06/12 10:00
PHST- 2002/06/12 10:00 [pubmed]
PHST- 2002/12/04 04:00 [medline]
PHST- 2002/06/12 10:00 [entrez]
AID - S0021-9258(20)74289-4 [pii]
AID - 10.1074/jbc.M201726200 [doi]
PST - ppublish
SO  - J Biol Chem. 2002 Sep 13;277(37):33978-86. doi: 10.1074/jbc.M201726200. Epub 2002 
      Jun 10.