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• W2747222786 abstract "Human umbilical cord blood constitutes a rich source of endothelial progenitor cells (EPC) capable of differentiation into functional mature endothelial cells. The endothelium is a key modulator in the pathogenesis of inflammatory vasculopathies such as atherosclerosis. Specifically, Interleukin-1 (IL-1) plays a major role in the proinflammatory activation of the endothelium and the propagation of the vascular inflammatory process. Aim of this study was to examine the potential of Interleukin-1 receptor antagonist (IL-1ra) - transduced cord blood-derived endothelial cells (CBEC) to inhibit the vascular inflammatory aspects involved in the pathogenesis of atherosclerosis.For this purpose, cord blood-derived CD34+ EPC were differentiated into CBEC and retrovirally transduced with the constructs pLXSN and pLXSN-icIL-1ra, respectively, and expanded up to 1020 cells during in vitro culture. Transgene expression of the intracellular isoform of IL-1ra was confirmed by RT-PCR and Western Blot and quantified by ELISA throughout the expansion period, demonstrating that transduced cells showed a stable expression of IL-1ra over at least 4 months of culture.In the following, the effect of transgenic icIL-1ra expression on distinct processes involved in inflammatory vasculopathies was investigated. CBEC gene expression analysis (real time RT-PCR) revealed that steady state levels of the adhesion receptor ICAM-1 and the proinflammatory mediators MCP-1 and thrombin receptor were significantly reduced in transgenic CBEC. Endothelial cell activation induced by monocytes, IL-1s and TNFα stimulation was assessed by flow cytometry and real time RT-PCR. We observed that monocyte-induced upregulation of ICAM-1, VCAM-1, and tissue factor was significantly inhibited in icIL-1ra-transduced CBEC. Furthermore, transgenic CBEC showed a decreased proinflammatory activation profile upon cytokine stimulation. Interestingly, the ectopically expressed icIL-1ra did not only reduce IL-1s-induced CBEC activation, but also influenced TNF-induced stimulation. In particular, TNF-induced ICAM-1 expression was reduced by icIL-1ra. Functionally, as assessed by flow chamber measurements and adhesion assays, expression of the icIL-1ra transgene reduced leukocyte rolling and adhesion to resting CBEC as well as IL-1s stimulated CBEC. Furthermore, transendothelial migration of monocytes through transgenic CBEC was markedly diminished as determined in an in vitro transmigration chamber.Our findings demonstrate that CBEC can reliably be isolated, transduced, and expanded into clinically relevant numbers. CBEC stably expressing icIL-1ra show an enforced protection against activation by proinflammatory stimuli and display a significantly reduced response in the decisive pathomechanisms of inflammatory vasculopathies - rolling, adhesion, and transmigration of leukocytes. Therefore, icIL-ra transgenic CBEC show potential to be used in the treatment of IL-1s- as well as TNFα-mediated inflammatory vasculopathies." @default.
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• W2747222786 date "2008-01-01" @default.
• W2747222786 modified "2023-09-23" @default.
• W2747222786 title "Inhibition of Inflammatory Vasculopathic Processes by Interleukin-1 Receptor Antagonist - transduced Endothelial Progenitor Cells" @default.
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