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Monocyte production of IL-12 is associated with the IL-28B genotype.

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posted on 2016-09-01, 04:55 authored by Benjamin 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

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 (A). NK cells from HCV(+) patients (n = 16) were co-cultured with R848 stimulated autologous monocytes and HuH7HCVReplicon cells in the presence or absence of anti-IL-12, anti-IL-18, or a combination of both mAbs. Then, IFN-γ production of CD56Bright NK cells was studied (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 (C). Next, R848-stimulated monocytes from HCV(+) patients that were stratified according to the IL-28B genotype (C/C, n = 7; C/T, n = 3; T/T, n = 6) were cultured with NK cells. Then, IFN-γ production of CD56Bright NK cells was tested following co-incubation with HuH7HCVreplicon in the presence of anti-IL-12 and/or anti-IL-18 (D). 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 (E).

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