Stably integrated luxCDABE for assessment of salmonella invasion kinetics

Kelly N. Flentie, Min Qi, Seth T. Gammon, Yasmin Razia, Felix Lui, Luciano Marpegan, Aashish Manglik, David Piwnica-Worms, Jeffrey S. McKinney

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

Salmonella Typhimurium is a common cause of gastroenteritis in humans and also localizes to neoplastic tumors in animals. Invasion of specific eukaryotic cells is a key mechanism of Salmonella interactions with host tissues. Early stages of gastrointestinal cell invasion are mediated by a Salmonella type III secretion system, powered by the adenosine triphosphatase invC. The aim of this work was to characterize the invC dependence of invasion kinetics into disparate eukaryotic cells traditionally used as models of gut epithelium or neoplasms. Thus, a nondestructive real-time assay was developed to report eukaryotic cell invasion kinetics using lux+ Salmonella that contain chromosomally integrated luxCDABE genes. Bioluminescence- based invasion assays using lux+ Salmonella exhibited inoculum dose-response correlation, distinguished invasion-competent from invasion-incompetent Salmonella, and discriminated relative Salmonella invasiveness in accordance with environmental conditions that induce invasion gene expression. In standard gentamicin protection assays, bioluminescence from lux+ Salmonella correlated with recovery of colony-forming units of internalized bacteria and could be visualized by bioluminescence microscopy. Furthermore, this assay distinguished invasion-competent from invasion-incompetent bacteria independent of gentamicin treatment in real time. Bioluminescence reported Salmonella invasion of disparate eukaryotic cell lines, including neoplastic melanoma, colon adenocarcinoma, and glioma cell lines used in animal models of malignancy. In each case, Salmonella invasion of eukaryotic cells was invC dependent.

Original languageEnglish (US)
Pages (from-to)222-233
Number of pages12
JournalMolecular Imaging
Volume7
Issue number5
DOIs
StatePublished - Sep 2008

Fingerprint

salmonella
Salmonella
Kinetics
Eukaryotic Cells
Bioluminescence
kinetics
bioluminescence
Assays
cells
Gentamicins
cultured cells
bacteria
Bacteria
Animals
inoculum
Cells
Cell Line
Neoplasms
neoplasms
adenosines

ASJC Scopus subject areas

  • Radiology Nuclear Medicine and imaging
  • Biotechnology
  • Molecular Medicine
  • Biomedical Engineering
  • Condensed Matter Physics

Cite this

Stably integrated luxCDABE for assessment of salmonella invasion kinetics. / Flentie, Kelly N.; Qi, Min; Gammon, Seth T.; Razia, Yasmin; Lui, Felix; Marpegan, Luciano; Manglik, Aashish; Piwnica-Worms, David; McKinney, Jeffrey S.

In: Molecular Imaging, Vol. 7, No. 5, 09.2008, p. 222-233.

Research output: Contribution to journalArticle

Flentie, KN, Qi, M, Gammon, ST, Razia, Y, Lui, F, Marpegan, L, Manglik, A, Piwnica-Worms, D & McKinney, JS 2008, 'Stably integrated luxCDABE for assessment of salmonella invasion kinetics', Molecular Imaging, vol. 7, no. 5, pp. 222-233. https://doi.org/10.2310/7290.2008.00024
Flentie KN, Qi M, Gammon ST, Razia Y, Lui F, Marpegan L et al. Stably integrated luxCDABE for assessment of salmonella invasion kinetics. Molecular Imaging. 2008 Sep;7(5):222-233. https://doi.org/10.2310/7290.2008.00024
Flentie, Kelly N. ; Qi, Min ; Gammon, Seth T. ; Razia, Yasmin ; Lui, Felix ; Marpegan, Luciano ; Manglik, Aashish ; Piwnica-Worms, David ; McKinney, Jeffrey S. / Stably integrated luxCDABE for assessment of salmonella invasion kinetics. In: Molecular Imaging. 2008 ; Vol. 7, No. 5. pp. 222-233.
@article{58936b6040fe46988d37148f408c5ad7,
title = "Stably integrated luxCDABE for assessment of salmonella invasion kinetics",
abstract = "Salmonella Typhimurium is a common cause of gastroenteritis in humans and also localizes to neoplastic tumors in animals. Invasion of specific eukaryotic cells is a key mechanism of Salmonella interactions with host tissues. Early stages of gastrointestinal cell invasion are mediated by a Salmonella type III secretion system, powered by the adenosine triphosphatase invC. The aim of this work was to characterize the invC dependence of invasion kinetics into disparate eukaryotic cells traditionally used as models of gut epithelium or neoplasms. Thus, a nondestructive real-time assay was developed to report eukaryotic cell invasion kinetics using lux+ Salmonella that contain chromosomally integrated luxCDABE genes. Bioluminescence- based invasion assays using lux+ Salmonella exhibited inoculum dose-response correlation, distinguished invasion-competent from invasion-incompetent Salmonella, and discriminated relative Salmonella invasiveness in accordance with environmental conditions that induce invasion gene expression. In standard gentamicin protection assays, bioluminescence from lux+ Salmonella correlated with recovery of colony-forming units of internalized bacteria and could be visualized by bioluminescence microscopy. Furthermore, this assay distinguished invasion-competent from invasion-incompetent bacteria independent of gentamicin treatment in real time. Bioluminescence reported Salmonella invasion of disparate eukaryotic cell lines, including neoplastic melanoma, colon adenocarcinoma, and glioma cell lines used in animal models of malignancy. In each case, Salmonella invasion of eukaryotic cells was invC dependent.",
author = "Flentie, {Kelly N.} and Min Qi and Gammon, {Seth T.} and Yasmin Razia and Felix Lui and Luciano Marpegan and Aashish Manglik and David Piwnica-Worms and McKinney, {Jeffrey S.}",
year = "2008",
month = "9",
doi = "10.2310/7290.2008.00024",
language = "English (US)",
volume = "7",
pages = "222--233",
journal = "Molecular Imaging",
issn = "1535-3508",
publisher = "Decker Publishing",
number = "5",

}

TY - JOUR

T1 - Stably integrated luxCDABE for assessment of salmonella invasion kinetics

AU - Flentie, Kelly N.

AU - Qi, Min

AU - Gammon, Seth T.

AU - Razia, Yasmin

AU - Lui, Felix

AU - Marpegan, Luciano

AU - Manglik, Aashish

AU - Piwnica-Worms, David

AU - McKinney, Jeffrey S.

PY - 2008/9

Y1 - 2008/9

N2 - Salmonella Typhimurium is a common cause of gastroenteritis in humans and also localizes to neoplastic tumors in animals. Invasion of specific eukaryotic cells is a key mechanism of Salmonella interactions with host tissues. Early stages of gastrointestinal cell invasion are mediated by a Salmonella type III secretion system, powered by the adenosine triphosphatase invC. The aim of this work was to characterize the invC dependence of invasion kinetics into disparate eukaryotic cells traditionally used as models of gut epithelium or neoplasms. Thus, a nondestructive real-time assay was developed to report eukaryotic cell invasion kinetics using lux+ Salmonella that contain chromosomally integrated luxCDABE genes. Bioluminescence- based invasion assays using lux+ Salmonella exhibited inoculum dose-response correlation, distinguished invasion-competent from invasion-incompetent Salmonella, and discriminated relative Salmonella invasiveness in accordance with environmental conditions that induce invasion gene expression. In standard gentamicin protection assays, bioluminescence from lux+ Salmonella correlated with recovery of colony-forming units of internalized bacteria and could be visualized by bioluminescence microscopy. Furthermore, this assay distinguished invasion-competent from invasion-incompetent bacteria independent of gentamicin treatment in real time. Bioluminescence reported Salmonella invasion of disparate eukaryotic cell lines, including neoplastic melanoma, colon adenocarcinoma, and glioma cell lines used in animal models of malignancy. In each case, Salmonella invasion of eukaryotic cells was invC dependent.

AB - Salmonella Typhimurium is a common cause of gastroenteritis in humans and also localizes to neoplastic tumors in animals. Invasion of specific eukaryotic cells is a key mechanism of Salmonella interactions with host tissues. Early stages of gastrointestinal cell invasion are mediated by a Salmonella type III secretion system, powered by the adenosine triphosphatase invC. The aim of this work was to characterize the invC dependence of invasion kinetics into disparate eukaryotic cells traditionally used as models of gut epithelium or neoplasms. Thus, a nondestructive real-time assay was developed to report eukaryotic cell invasion kinetics using lux+ Salmonella that contain chromosomally integrated luxCDABE genes. Bioluminescence- based invasion assays using lux+ Salmonella exhibited inoculum dose-response correlation, distinguished invasion-competent from invasion-incompetent Salmonella, and discriminated relative Salmonella invasiveness in accordance with environmental conditions that induce invasion gene expression. In standard gentamicin protection assays, bioluminescence from lux+ Salmonella correlated with recovery of colony-forming units of internalized bacteria and could be visualized by bioluminescence microscopy. Furthermore, this assay distinguished invasion-competent from invasion-incompetent bacteria independent of gentamicin treatment in real time. Bioluminescence reported Salmonella invasion of disparate eukaryotic cell lines, including neoplastic melanoma, colon adenocarcinoma, and glioma cell lines used in animal models of malignancy. In each case, Salmonella invasion of eukaryotic cells was invC dependent.

UR - http://www.scopus.com/inward/record.url?scp=61549105587&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=61549105587&partnerID=8YFLogxK

U2 - 10.2310/7290.2008.00024

DO - 10.2310/7290.2008.00024

M3 - Article

C2 - 19123992

AN - SCOPUS:61549105587

VL - 7

SP - 222

EP - 233

JO - Molecular Imaging

JF - Molecular Imaging

SN - 1535-3508

IS - 5

ER -