posted on 2016-09-01, 04:55authored byBenjamin Krämer, Claudia Finnemann, Beatriz Sastre, Philipp Lutz, Andreas Glässner, Franziska Wolter, Felix Goeser, Pavlos Kokordelis, Dominik Kaczmarek, Hans-Dieter Nischalke, Christian P. Strassburg, Ulrich Spengler, Jacob Nattermann
<p>Purified monocytes from HCV(+) patients were stimulated with R848 and then tested for expression of IL-12-p70 (n = 12), IL-12-p40 (n = 19), and IL-18 (n = 15), respectively <b>(A).</b> NK cells from HCV(+) patients (n = 16) were co-cultured with R848 stimulated autologous monocytes and HuH7<sub>HCV</sub>Replicon cells in the presence or absence of anti-IL-12, anti-IL-18, or a combination of both mAbs. Then, IFN-γ production of CD56<sup>Bright</sup> NK cells was studied <b>(B)</b>. Purified monocytes from HCV(+) patients were stimulated with R848. After 16h the resulting supernatants were harvested and tested for concentrations of IL-12-p40 (C/C, n = 7; C/T, n = 8; T/T, n = 4), IL-12-p70 (C/C, n = 5; C/T, n = 3; T/T, n = 4), and IL-18 (C/C, n = 7; C/T, n = 4; T/T, n = 4), respectively, by ELISA <b>(C)</b>. Next, R848-stimulated monocytes from HCV(+) patients that were stratified according to the <i>IL-28B</i> genotype (C/C, n = 7; C/T, n = 3; T/T, n = 6) were cultured with NK cells. Then, IFN-γ production of CD56<sup>Bright</sup> NK cells was tested following co-incubation with HuH7<sub>HCV</sub>replicon in the presence of anti-IL-12 and/or anti-IL-18 <b>(D)</b>. As a further control, purified monocytes from healthy donors were stimulated with R848. Then, concentrations of IL-12-p40 (C/C, n = 6; C/T, n = 5; T/T, n = 3; left graph) and IL-18 (C/C, n = 6; C/T, n = 7; T/T, n = 4; right graph) were analysed by ELISA <b>(E)</b>.</p>