Phenotypic and Genetic Evaluation of the Influence of Pseudomonas aeruginosa Culture Fractions on the Human Mesenchymal Stem Cells Viability, Apoptotic Pathways and Cytokine Profile.
The objective of this study was to investigate the effects of P. aeruginosa PAO1 cellular and soluble culture fractions on human mesenchymal stem cells (MSCs) death signaling pathways and cytokine profile. The bone marrow isolated MSCs, incubated for different periods of time with one of the three P. aeruginosa PAO1 culture fractions, i.e. low density whole cultures, heat inactivated bacterial cultures sediments and sterile supernatants, were submitted to the following assays: i) fluorescence microscopy evaluation of cellular morphology and viability; ii) bax, caspase 9, relA and bcl-2 genes expression analysis by qRT-PCR; and iii) quantification of the level of IL-1β, IL-6, IL-8 and IL-10 cytokines released in the MSCs supernatants determined by ELISA. Results were statistically analyzed using the GraphPad In Stat software. The PAO1 whole cultures exhibited the most relevant influences, impacting on MSCs morphology and viability, interfering with apoptotic pathways and significantly stimulating the production of IL-1β and IL-10, while decreasing the production of IL-6 and IL-8. The culture supernatants increased the production of IL-1β and reduced the secretion of all other tested cytokines, while heat-inactivated bacterial cells significantly stimulated both IL-1β and IL-10 production. These data could suggest that in vivo, the fate of P. aeruginosa infection depends on the proportion between different bacterial culture fractions (i.e. the number of viable bacterial cells, the number of dead cells and the amount of bacterial soluble products accumulated locally) that could be influenced by the initial infective dose, by the host defense mechanisms, and also by the administered antimicrobial treatment that may thus interfere with the evolution and magnitude of the induced lesions.
Department of Microbiology and Immunology, Faculty of Biology, University of Bucharest, Intrarea Portocalelor 1-3, 060101, Sector 6, Bucharest, Romania.
Alina Maria
AM
Holban
Coralia
C
Bleotu
Mariana Carmen
MC
Chifiriuc
Veronica
V
Lazar
D017209:Apoptosis / Q000187:drug effects*; D001854:Bone Marrow Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D053453:Caspase 9 / Q000235:genetics / Q000276:immunology; D002470:Cell Survival / Q000187:drug effects; D005591:Chemical Fractionation; D017077:Culture Media, Conditioned / Q000494:pharmacology*; D005786:Gene Expression Regulation; D054884:Host-Pathogen Interactions; D006358:Hot Temperature; D006801:Humans; D016753:Interleukin-10 / Q000819:agonists / Q000235:genetics / Q000276:immunology; D053583:Interleukin-1beta / Q000819:agonists / Q000235:genetics / Q000276:immunology; D015850:Interleukin-6 / Q000235:genetics / Q000276:immunology; D016209:Interleukin-8 / Q000037:antagonists & inhibitors / Q000235:genetics / Q000276:immunology; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D061251:Primary Cell Culture; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000235:genetics / Q000276:immunology; D011550:Pseudomonas aeruginosa / Q000737:chemistry* / Q000254:growth & development / Q000472:pathogenicity; D015398:Signal Transduction; D051996:Transcription Factor RelA / Q000235:genetics / Q000276:immunology; D051028:bcl-2-Associated X Protein / Q000235:genetics / Q000276:immunology; D000093242:Interleukin-6 Inhibitors
D053453:Caspase 9; D017077:Culture Media, Conditioned; D016753:Interleukin-10; D053583:Interleukin-1beta; D015850:Interleukin-6; D016209:Interleukin-8; D019253:Proto-Oncogene Proteins c-bcl-2; D051996:Transcription Factor RelA; D051028:bcl-2-Associated X Protein; D000093242:Interleukin-6 Inhibitors; C494826:BAX protein, human; C000595707:BCL2 protein, human; C508609:IL10 protein, human; C508585:IL1B protein, human; C508600:IL6 protein, human; C498143:RELA protein, human; C506433:CASP9 protein, human
Apoptosis; Pseudomonas aeruginosa.; interleukins; mesenchymal stem cells; pathogenesis