title: Phenotypic pattern-based assay for dynamically monitoring host cellular responses to Salmonella infections.
abstract: The interaction between mammalian host cells and bacteria is a dynamic process, and the underlying pathologic mechanisms are poorly characterized. Limited information describing the host-bacterial interaction is based mainly on studies using label-based endpoint assays that detect changes in cell behavior at a given time point, yielding incomplete information. In this paper, a novel, label-free, real-time cell-detection system based on electronic impedance sensor technology was adapted to dynamically monitor the entire process of intestinal epithelial cells response to Salmonella infection. Changes in cell morphology and attachment were quantitatively and continuously recorded following infection. The resulting impedance-based time-dependent cell response profiles (TCRPs) were compared to standard assays and showed good correlation and sensitivity. Biochemical assays further suggested that TCRPs were correlated with cytoskeleton-associated morphological dynamics, which can be largely attenuated by inhibitions of actin and microtubule polymerization. Collectively, our data indicate that cell-electrode impedance measurements not only provide a novel, real-time, label-free method for investigating bacterial infection but also help advance our understanding of host responses in a more physiological and continuous manner that is beyond the scope of current endpoint assays.
authors.affiliation: State Key Laboratory for Diagnosis and Treatment of Infectious Diseases, The First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, China.
authors.forename: Xiaozhou
authors.initials: X
authors.lastname: Mou
authors.forename: Shuying
authors.initials: S
authors.lastname: Wan
authors.forename: Yifei
authors.initials: Y
authors.lastname: Li
authors.forename: Shanshan
authors.initials: S
authors.lastname: Zhang
authors.forename: Mingjiao
authors.initials: M
authors.lastname: Sun
authors.forename: Fanglong
authors.initials: F
authors.lastname: Liu
authors.forename: Huiying
authors.initials: H
authors.lastname: Fu
authors.forename: Xue
authors.initials: X
authors.lastname: Zhang
authors.forename: Haiying
authors.initials: H
authors.lastname: Liu
authors.forename: Qian
authors.initials: Q
authors.lastname: Cao
authors.forename: Yuehai
authors.initials: Y
authors.lastname: Ke
authors.forename: Charlie
authors.initials: C
authors.lastname: Xiang
mesh_terms: D019073:HT29 Cells; D006801:Humans; D010641:Phenotype; D012480:Salmonella Infections / Q000276:immunology* / Q000473:pathology
references.citation: Proc Natl Acad Sci U S A. 1999 Sep 28;96(20):11578-83
references.pmid: 10500219
references.citation: J Immunol. 2000 Sep 15;165(6):2955-61
references.pmid: 10975802
references.citation: Nat Rev Microbiol. 2008 Jan;6(1):53-66
references.pmid: 18026123
references.citation: Anal Chem. 2010 Aug 1;82(15):6495-503
references.pmid: 20617823
references.citation: Assay Drug Dev Technol. 2006 Oct;4(5):545-53
references.pmid: 17115925
references.citation: Nature. 2000 Oct 12;407(6805):770-6
references.pmid: 11048727
references.citation: Nature. 2001 Apr 26;410(6832):1099-103
references.pmid: 11323673
references.citation: Chem Res Toxicol. 2005 Feb;18(2):154-61
references.pmid: 15720119
references.citation: J Clin Invest. 1998 Nov 15;102(10):1815-23
references.pmid: 9819367
references.citation: J Cell Biol. 1988 Jul;107(1):221-30
references.pmid: 3292541
references.citation: Nature. 1993 Dec 9;366(6455):591-2
references.pmid: 8255299
references.citation: J Biol Chem. 2002 Dec 27;277(52):51025-32
references.pmid: 12381718
references.citation: Mol Microbiol. 2000 Oct;38(1):31-40
references.pmid: 11029688
references.citation: Methods Mol Biol. 2008;431:201-11
references.pmid: 18287758
references.citation: Proc Natl Acad Sci U S A. 2000 Aug 1;97(16):8754-61
references.pmid: 10922031
references.citation: J Microbiol Methods. 2009 Jul;78(1):97-100
references.pmid: 19393695
references.citation: J Biomol Screen. 2005 Dec;10(8):795-805
references.pmid: 16234347
references.citation: Chem Biol. 2009 Jul 31;16(7):712-23
references.pmid: 19635408
references.citation: Proc Natl Acad Sci U S A. 2001 May 8;98(10):5850-5
references.pmid: 11320214
references.citation: Assay Drug Dev Technol. 2004 Aug;2(4):363-72
references.pmid: 15357917
references.citation: J Cell Sci. 1991 Jun;99 ( Pt 2):283-96
references.pmid: 1909337
references.citation: J Biomol Screen. 2006 Sep;11(6):634-43
references.pmid: 16858006
references.citation: Genome Biol. 2003;4(1):R2
references.pmid: 12537547
references.citation: Clin Microbiol Rev. 2008 Jan;21(1):134-56
references.pmid: 18202440
references.citation: J Microbiol Methods. 2011 Jan;84(1):101-8
references.pmid: 21078346
references.citation: Curr Opin Microbiol. 2000 Feb;3(1):54-9
references.pmid: 10679413
references.citation: Science. 1996 Dec 20;274(5295):2115-8
references.pmid: 8953049
references.citation: J Clin Microbiol. 2010 Nov;48(11):4129-34
references.pmid: 20720023
references.citation: Int J Med Microbiol. 2003 Apr;293(1):41-54
references.pmid: 12755365
references.citation: Annu Rev Med. 2001;52:259-74
references.pmid: 11160778
references.citation: Anal Chem. 2011 Sep 1;83(17):6518-26
references.pmid: 21793491
references.citation: PLoS Pathog. 2010 Apr 29;6(4):e1000874
references.pmid: 20442866
references.citation: Microbes Infect. 2004 Apr;6(4):398-405
references.pmid: 15101397
references.citation: Mol Cell Biol. 2007 Oct;27(19):6706-17
references.pmid: 17646384
references.citation: Trends Biotechnol. 2010 May;28(5):237-45
references.pmid: 20346526
references.citation: Trends Microbiol. 2005 Oct;13(10):497-503
references.pmid: 16140013
references.citation: PLoS Pathog. 2008 Apr 04;4(4):e1000037
references.pmid: 18389058
references.citation: Analyst. 2008 May;133(5):643-8
references.pmid: 18427686
references.citation: Oncogene. 2007 Jul 26;26(34):4951-60
references.pmid: 17310989
references.citation: J Immunol Methods. 2004 Sep;292(1-2):195-205
references.pmid: 15350524
references.citation: Am J Physiol Gastrointest Liver Physiol. 2006 Jan;290(1):G96-G108
references.pmid: 16179598
references.citation: Nat Rev Mol Cell Biol. 2003 May;4(5):385-96
references.pmid: 12728272
references.citation: Proc Natl Acad Sci U S A. 1984 Jun;81(12):3761-4
references.pmid: 6587391
references.citation: Microbes Infect. 2001 Nov-Dec;3(14-15):1321-6
references.pmid: 11755421
references.citation: Proc Natl Acad Sci U S A. 1996 Sep 3;93(18):9833-8
references.pmid: 8790417
references.citation: Anal Chem. 2006 Jan 1;78(1):35-43
references.pmid: 16383308