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10.1371/journal.pone.0033602 http://scihub22266oqcxt.onion/10.1371/journal.pone.0033602 22438961!3305306!22438961     free     free     free Warning: file_get_contents(https://eutils.ncbi.nlm.nih.gov/entrez/eutils/elink.fcgi?dbfrom=pubmed&id=22438961&cmd=llinks): Failed to open stream: HTTP request failed! 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Green fluorescent protein (GFP) color reporter gene visualizes parvovirus B19 non-structural segment 1 (NS1) transfected endothelial modification |pdf=|usr=}}{{22438961}} gab.com Text Green fluorescent protein (GFP) color reporter gene visualizes parvovirus B19 non-structural segment 1 (NS1) transfected endothelial modification JO: PLoS One http://www.kidney.de/pget.php?&tw=1&pmid=22438961 #FOAvip BACKGROUND: Human Parvovirus B19 (PVB19) has been associated with myocarditis putative due to endothelial infection. Whether PVB19 infects endothelial cells and causes a modification of endothelial function and inflammation and, thus, disturbance of microcirculation has not been elucidated and could not be visualized so far. METHODS AND FINDINGS: To examine the PVB19-induced endothelial modification, we used green fluorescent protein (GFP) color reporter gene in the non-structural segment 1 (NS1) of PVB19. NS1-GFP-PVB19 or GFP plasmid as control were transfected in an endothelial-like cell line (ECV304). The endothelial surface expression of intercellular-adhesion molecule-1 (CD54/ICAM-1) and extracellular matrix metalloproteinase inducer (EMMPRIN/CD147) were evaluated by flow cytometry after NS-1-GFP or control-GFP transfection. To evaluate platelet adhesion on NS-1 transfected ECs, we performed a dynamic adhesion assay (flow chamber). NS-1 transfection causes endothelial activation and enhanced expression of ICAM-1 (CD54: mean +/- standard deviation: NS1-GFP vs. control-GFP: 85.3 +/- 11.2 vs. 61.6 +/- 8.1; P<0.05) and induces endothelial expression of EMMPRIN/CD147 (CD147: mean +/- SEM: NS1-GFP vs. control-GFP: 114 +/- 15.3 vs. 80 +/- 0.91; P<0.05) compared to control-GFP transfected cells. Dynamic adhesion assays showed that adhesion of platelets is significantly enhanced on NS1 transfected ECs when compared to control-GFP (P<0.05). The transfection of ECs was verified simultaneously through flow cytometry, immunofluorescence microscopy and polymerase chain reaction (PCR) analysis. CONCLUSIONS: GFP color reporter gene shows transfection of ECs and may help to visualize NS1-PVB19 induced endothelial activation and platelet adhesion as well as an enhanced monocyte adhesion directly, providing in vitro evidence of possible microcirculatory dysfunction in PVB19-induced myocarditis and, thus, myocardial tissue damage. Twit Text FOAVip Green fluorescent protein (GFP) color reporter gene visualizes parvovirus B19 non-structural segment 1 (NS1) transfected endothelial modification ????PLoS One http://www.kidney.de/pget.php?&tw=1&pmid=22438961 #FOAvip Twit Text # Free Paper on # # # # # LINK http://www.kidney.de/pget.php?&tw=1&pmid=22438961 #FOAvip English Wikipedia <ref>{{Cite pmid|22438961}}</ref> Green fluorescent protein (GFP) color reporter gene visualizes parvovirus B19 non-structural segment 1 (NS1) transfected endothelial modification #MMPMID22438961 Wurster T; Polzelbauer C; Schonberger T; Paul A; Seizer P; Stellos K; Schuster A; Botnar RM; Gawaz M; Bigalke B PLoS One 2012[]; 7 (3): e33602 PMID22438961show ga BACKGROUND: Human Parvovirus B19 (PVB19) has been associated with myocarditis putative due to endothelial infection. Whether PVB19 infects endothelial cells and causes a modification of endothelial function and inflammation and, thus, disturbance of microcirculation has not been elucidated and could not be visualized so far. METHODS AND FINDINGS: To examine the PVB19-induced endothelial modification, we used green fluorescent protein (GFP) color reporter gene in the non-structural segment 1 (NS1) of PVB19. NS1-GFP-PVB19 or GFP plasmid as control were transfected in an endothelial-like cell line (ECV304). The endothelial surface expression of intercellular-adhesion molecule-1 (CD54/ICAM-1) and extracellular matrix metalloproteinase inducer (EMMPRIN/CD147) were evaluated by flow cytometry after NS-1-GFP or control-GFP transfection. To evaluate platelet adhesion on NS-1 transfected ECs, we performed a dynamic adhesion assay (flow chamber). NS-1 transfection causes endothelial activation and enhanced expression of ICAM-1 (CD54: mean +/- standard deviation: NS1-GFP vs. control-GFP: 85.3 +/- 11.2 vs. 61.6 +/- 8.1; P<0.05) and induces endothelial expression of EMMPRIN/CD147 (CD147: mean +/- SEM: NS1-GFP vs. control-GFP: 114 +/- 15.3 vs. 80 +/- 0.91; P<0.05) compared to control-GFP transfected cells. Dynamic adhesion assays showed that adhesion of platelets is significantly enhanced on NS1 transfected ECs when compared to control-GFP (P<0.05). The transfection of ECs was verified simultaneously through flow cytometry, immunofluorescence microscopy and polymerase chain reaction (PCR) analysis. CONCLUSIONS: GFP color reporter gene shows transfection of ECs and may help to visualize NS1-PVB19 induced endothelial activation and platelet adhesion as well as an enhanced monocyte adhesion directly, providing in vitro evidence of possible microcirculatory dysfunction in PVB19-induced myocarditis and, thus, myocardial tissue damage. |Base Sequence[MESH] |Basigin/metabolism[MESH] |Cell Line[MESH] |DNA, Viral/genetics[MESH] |Endothelial Cells/metabolism/virology[MESH] |Genes, Reporter[MESH] |Green Fluorescent Proteins/*genetics[MESH] |Humans[MESH] |Intercellular Adhesion Molecule-1/metabolism[MESH] |Microvessels/virology[MESH] |Myocarditis/virology[MESH] |Parvoviridae Infections/virology[MESH] |Parvovirus B19, Human/*genetics/metabolism/pathogenicity[MESH] |Platelet Adhesiveness[MESH] |Recombinant Fusion Proteins/genetics/metabolism[MESH] |Transfection[MESH]Green fluorescent protein (GFP) color reporter gene visualizes parvovi(epmc).pdf DeepDyve Pubget Overpricing