Effects of Helicobacter pylori on intracellular Ca2+ signaling in normal human gastric mucous epithelial cells

Katie L. Marlink, Kathy D. Bacon, Brett Sheppard, Hassan Ashktorab, Duane T. Smoot, Timothy L. Cover, Clifford Deveney, Michael Rutten

Research output: Contribution to journalArticle

26 Citations (Scopus)

Abstract

In stomach, Helicobacter pylori (Hp) adheres to gastric mucous epithelial cells (GMEC) and initiates several different signal transduction events. Alteration of intracellular Ca2+ concentration ([Ca2+]i) is an important signaling mechanism in numerous bacteria-host model systems. Changes in [Ca2+]i induced by Hp in normal human GMEC have not yet been described; therefore, we examined effects of Hp on [Ca2+]i in normal human GMEC and a nontransformed GMEC line (HFE-145). Cultured cells were grown on glass slides, porous filters, or 96-well plates and loaded with fura 2 or fluo 4. Hp wild-type strain 60190 and vacA-, cagA-, and picB-/cagE- isogenic mutants were incubated with cells. Changes in [Ca2+]i were recorded with a fluorimeter or fluorescence plate reader. Wild-type Hp produced dose-dependent biphasic transient [Ca2+]i peak and plateau changes in both cell lines. Hp vacA- isogenic mutant produced changes in [Ca2+]i similar to those produced by wild type. Compared with wild type, cagA- and picB-/cagE- isogenic mutants produced lower peak changes and did not generate a plateau change. Preloading cultures with intracellular Ca2+ chelator BAPTA blocked all Hp-induced [Ca2+]i changes. Thapsigargin pretreatment of cultures to release Ca2+ from internal stores reduced peak change. Extracellular Ca2+ removal reduced plateau response. Hp-induced peak response was sensitive to G proteins and PLC inhibitors. Hp-induced plateau change was sensitive to G protein inhibitors, src kinases, and PLA2. These findings are the first to show that H. pylori alters [Ca2+]i in normal GMEC through a Ca2+ release/influx mechanism that depends on expression of caga and picB/cagE genes.

Original languageEnglish (US)
JournalAmerican Journal of Physiology - Gastrointestinal and Liver Physiology
Volume285
Issue number1 48-1
StatePublished - Jul 1 2003

Fingerprint

Helicobacter pylori
Stomach
Epithelial Cells
GTP-Binding Proteins
Cell Line
Thapsigargin
src-Family Kinases
Fura-2
Chelating Agents
Glass
Cultured Cells
Signal Transduction
Fluorescence
Bacteria

Keywords

  • Bacteria
  • cagA
  • Cell culture
  • Fluo 4
  • Fura 2
  • G protein
  • Genistein
  • Herbimycin
  • Immunofluorescence
  • picB/cagE
  • Signal transduction
  • Stomach
  • Thapsigargin
  • vacA

ASJC Scopus subject areas

  • Gastroenterology
  • Physiology

Cite this

Effects of Helicobacter pylori on intracellular Ca2+ signaling in normal human gastric mucous epithelial cells. / Marlink, Katie L.; Bacon, Kathy D.; Sheppard, Brett; Ashktorab, Hassan; Smoot, Duane T.; Cover, Timothy L.; Deveney, Clifford; Rutten, Michael.

In: American Journal of Physiology - Gastrointestinal and Liver Physiology, Vol. 285, No. 1 48-1, 01.07.2003.

Research output: Contribution to journalArticle

Marlink, Katie L. ; Bacon, Kathy D. ; Sheppard, Brett ; Ashktorab, Hassan ; Smoot, Duane T. ; Cover, Timothy L. ; Deveney, Clifford ; Rutten, Michael. / Effects of Helicobacter pylori on intracellular Ca2+ signaling in normal human gastric mucous epithelial cells. In: American Journal of Physiology - Gastrointestinal and Liver Physiology. 2003 ; Vol. 285, No. 1 48-1.
@article{50552eab9d214755bc4c79c04d65c4cb,
title = "Effects of Helicobacter pylori on intracellular Ca2+ signaling in normal human gastric mucous epithelial cells",
abstract = "In stomach, Helicobacter pylori (Hp) adheres to gastric mucous epithelial cells (GMEC) and initiates several different signal transduction events. Alteration of intracellular Ca2+ concentration ([Ca2+]i) is an important signaling mechanism in numerous bacteria-host model systems. Changes in [Ca2+]i induced by Hp in normal human GMEC have not yet been described; therefore, we examined effects of Hp on [Ca2+]i in normal human GMEC and a nontransformed GMEC line (HFE-145). Cultured cells were grown on glass slides, porous filters, or 96-well plates and loaded with fura 2 or fluo 4. Hp wild-type strain 60190 and vacA-, cagA-, and picB-/cagE- isogenic mutants were incubated with cells. Changes in [Ca2+]i were recorded with a fluorimeter or fluorescence plate reader. Wild-type Hp produced dose-dependent biphasic transient [Ca2+]i peak and plateau changes in both cell lines. Hp vacA- isogenic mutant produced changes in [Ca2+]i similar to those produced by wild type. Compared with wild type, cagA- and picB-/cagE- isogenic mutants produced lower peak changes and did not generate a plateau change. Preloading cultures with intracellular Ca2+ chelator BAPTA blocked all Hp-induced [Ca2+]i changes. Thapsigargin pretreatment of cultures to release Ca2+ from internal stores reduced peak change. Extracellular Ca2+ removal reduced plateau response. Hp-induced peak response was sensitive to G proteins and PLC inhibitors. Hp-induced plateau change was sensitive to G protein inhibitors, src kinases, and PLA2. These findings are the first to show that H. pylori alters [Ca2+]i in normal GMEC through a Ca2+ release/influx mechanism that depends on expression of caga and picB/cagE genes.",
keywords = "Bacteria, cagA, Cell culture, Fluo 4, Fura 2, G protein, Genistein, Herbimycin, Immunofluorescence, picB/cagE, Signal transduction, Stomach, Thapsigargin, vacA",
author = "Marlink, {Katie L.} and Bacon, {Kathy D.} and Brett Sheppard and Hassan Ashktorab and Smoot, {Duane T.} and Cover, {Timothy L.} and Clifford Deveney and Michael Rutten",
year = "2003",
month = "7",
day = "1",
language = "English (US)",
volume = "285",
journal = "American Journal of Physiology - Renal Fluid and Electrolyte Physiology",
issn = "1931-857X",
publisher = "American Physiological Society",
number = "1 48-1",

}

TY - JOUR

T1 - Effects of Helicobacter pylori on intracellular Ca2+ signaling in normal human gastric mucous epithelial cells

AU - Marlink, Katie L.

AU - Bacon, Kathy D.

AU - Sheppard, Brett

AU - Ashktorab, Hassan

AU - Smoot, Duane T.

AU - Cover, Timothy L.

AU - Deveney, Clifford

AU - Rutten, Michael

PY - 2003/7/1

Y1 - 2003/7/1

N2 - In stomach, Helicobacter pylori (Hp) adheres to gastric mucous epithelial cells (GMEC) and initiates several different signal transduction events. Alteration of intracellular Ca2+ concentration ([Ca2+]i) is an important signaling mechanism in numerous bacteria-host model systems. Changes in [Ca2+]i induced by Hp in normal human GMEC have not yet been described; therefore, we examined effects of Hp on [Ca2+]i in normal human GMEC and a nontransformed GMEC line (HFE-145). Cultured cells were grown on glass slides, porous filters, or 96-well plates and loaded with fura 2 or fluo 4. Hp wild-type strain 60190 and vacA-, cagA-, and picB-/cagE- isogenic mutants were incubated with cells. Changes in [Ca2+]i were recorded with a fluorimeter or fluorescence plate reader. Wild-type Hp produced dose-dependent biphasic transient [Ca2+]i peak and plateau changes in both cell lines. Hp vacA- isogenic mutant produced changes in [Ca2+]i similar to those produced by wild type. Compared with wild type, cagA- and picB-/cagE- isogenic mutants produced lower peak changes and did not generate a plateau change. Preloading cultures with intracellular Ca2+ chelator BAPTA blocked all Hp-induced [Ca2+]i changes. Thapsigargin pretreatment of cultures to release Ca2+ from internal stores reduced peak change. Extracellular Ca2+ removal reduced plateau response. Hp-induced peak response was sensitive to G proteins and PLC inhibitors. Hp-induced plateau change was sensitive to G protein inhibitors, src kinases, and PLA2. These findings are the first to show that H. pylori alters [Ca2+]i in normal GMEC through a Ca2+ release/influx mechanism that depends on expression of caga and picB/cagE genes.

AB - In stomach, Helicobacter pylori (Hp) adheres to gastric mucous epithelial cells (GMEC) and initiates several different signal transduction events. Alteration of intracellular Ca2+ concentration ([Ca2+]i) is an important signaling mechanism in numerous bacteria-host model systems. Changes in [Ca2+]i induced by Hp in normal human GMEC have not yet been described; therefore, we examined effects of Hp on [Ca2+]i in normal human GMEC and a nontransformed GMEC line (HFE-145). Cultured cells were grown on glass slides, porous filters, or 96-well plates and loaded with fura 2 or fluo 4. Hp wild-type strain 60190 and vacA-, cagA-, and picB-/cagE- isogenic mutants were incubated with cells. Changes in [Ca2+]i were recorded with a fluorimeter or fluorescence plate reader. Wild-type Hp produced dose-dependent biphasic transient [Ca2+]i peak and plateau changes in both cell lines. Hp vacA- isogenic mutant produced changes in [Ca2+]i similar to those produced by wild type. Compared with wild type, cagA- and picB-/cagE- isogenic mutants produced lower peak changes and did not generate a plateau change. Preloading cultures with intracellular Ca2+ chelator BAPTA blocked all Hp-induced [Ca2+]i changes. Thapsigargin pretreatment of cultures to release Ca2+ from internal stores reduced peak change. Extracellular Ca2+ removal reduced plateau response. Hp-induced peak response was sensitive to G proteins and PLC inhibitors. Hp-induced plateau change was sensitive to G protein inhibitors, src kinases, and PLA2. These findings are the first to show that H. pylori alters [Ca2+]i in normal GMEC through a Ca2+ release/influx mechanism that depends on expression of caga and picB/cagE genes.

KW - Bacteria

KW - cagA

KW - Cell culture

KW - Fluo 4

KW - Fura 2

KW - G protein

KW - Genistein

KW - Herbimycin

KW - Immunofluorescence

KW - picB/cagE

KW - Signal transduction

KW - Stomach

KW - Thapsigargin

KW - vacA

UR - http://www.scopus.com/inward/record.url?scp=0037663735&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0037663735&partnerID=8YFLogxK

M3 - Article

VL - 285

JO - American Journal of Physiology - Renal Fluid and Electrolyte Physiology

JF - American Journal of Physiology - Renal Fluid and Electrolyte Physiology

SN - 1931-857X

IS - 1 48-1

ER -