Effects of Helicobacter hepaticus on the proteome of HEp-2 cells.
Helicobacter hepaticus infects the bowel and biliary tree of several animals, producing inflammation. Colonisation of mouse livers can induce hepatocellular carcinomas. The effects of H. hepaticus on the proliferation and global protein expression of human HEp-2 cells were studied by examining the changes in the protein profiles of cells exposed to the bacterium. HEp-2 cells were grown for four days under a microaerobic atmosphere or under the same conditions in co-cultures with H. hepaticus at various inoculum densities. Enlargement, distension and elongation of HEp-2 cells were observed in co-cultures with H. hepaticus. The number of live cells declined by only an order of magnitude at bacterial inocula of approximately 10(9)cfu/ml, but were reduced to less than 10(3)cells/ml at approximately 10(10)cfu/ml bacteria inocula. Protein expression by HEp-2 cells was investigated employing two-dimensional gel electrophoresis. In cells grown with or without bacteria, 17 differentially expressed proteins were identified by tandem mass spectrometry. These proteins participated in several biological functions including amino acid metabolism, cell growth and proliferation, stress response, protein translation and modification, etc. The onset of a catastrophic killing of HEp-2 cells at a bacterial density of approximately 10(9)cfu/ml suggested a multimodal action for H. hepaticus infection, and the modulation of the expression of proteins involved in different biological functions showed that the presence of H. hepaticus has broad effects on the physiology of HEp-2 cells.
Okoli
Arinze S
AS
School of Medical Sciences, The University of New South Wales, Sydney, NSW 2052, Australia.
Fox
Edward M
EM
Raftery
Mark J
MJ
Mendz
George L
GL
D000818:Animals; D002460:Cell Line; D049109:Cell Proliferation; D048430:Cell Shape / Q000502:physiology; D015180:Electrophoresis, Gel, Two-Dimensional; D016481:Helicobacter Infections / Q000378:metabolism; D044923:Helicobacter hepaticus / Q000502:physiology*; D006801:Humans; D020543:Proteome / Q000378:metabolism*; D053719:Tandem Mass Spectrometry
D020543:Proteome