Abstract
Interleukin-10 (IL-10) expression was triggered by electroporation in the human monocytic cell line U937. IL-10 protein was detected in the cell supernatant 8 h after pulse application and reached maximal concentration after 24 h. The protein yield was depended on both pulse duration and electric field strength. Electroporation of the cells in calcium free medium decreased IL-10 yield by 80%. Moreover, the presence of EGTA in the electroporation medium was capable to decrease IL-10 protein production in a dose-dependent manner. In comparison to the housekeeping gene glyceraldehyde-3-phosphodehydrogenase (GAPDH), a similar effect was shown at the IL-10 mRNA level as demonstrated by quantitative reverse transcriptase polymerase-chain reaction (RT-PCR). Furthermore the calmodulin antagonist W7 inhibited IL-10 protein production triggered by electroporation. Additionally, a luciferase reporter gene study revealed that the IL-10 promoter was activated by electroporation. W7 was capable to decrease the IL-10 promoter activity. The data strongly suggest that elevation of intracellular calcium concentration and subsequent formation of a calcium/calmodulin complex is necessary for IL-10 expression in monocytes. In conclusion, the opportunity to induce the antiinflammatory cytokine IL-10 by electroporation could lead to the development of new therapeutic strategies.
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