Fig 1: TNF-a, IL-6, and IL-10 are upregulated in PBMC and monocytes after ß-glucan stimulation. (A) PBMC were cultured with or without ß-glucan (Macrogard or Curdlan at 10 µg/ml). After 4 h, primed PBMC were collected and used for qPCR analysis. (B–E) PBMC, CD14+, and CD14- populations were cultured with or without ß-glucan (Macrogard or Curdlan at 10 µg/ml) for 16 h. Culture supernatant was collected and analyzed by ELISA for presence of cytokines. *p < 0.05; **p < 0.01; ****p < 0.0001.
Fig 2: IL-10 plays a cardinal role in the indirect stimulation of NK cell cytotoxic capacity upon ß-glucan priming of monocytes. Freshly isolated NK cells were cultured with recombinant porcine TNF-a (A), IL-10 (D), and IL-6 (G) at different concentrations for 16 h. Afterwards, primed NK cells were co-cultured with K562 target cells for 4 h to assess cytotoxic activity. PBMC were cultured with or without ß-glucan (Macrogard or Curdlan at 10 µg/ml) for 16 h. Afterwards, supernatant was collected and used for immunoprecipitation (IP) with a specific anti-cytokine antibody for TNF-a, IL-10, or IL-6 or without antibody as negative control. Afterwards, efficiency of IP was controlled by ELISA (B,E,H). Supernatant obtained after IP was added to freshly isolated NK cells to define the role of TNF-a (C), IL-10 (F), and IL-6 (I) in the indirect stimulation of NK cells upon ß-glucan priming of monocytes. After 16 h of incubation, primed NK cells were co-cultured with K562 target cells for 4 h to assess cytotoxic activity. *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001; ns, not significant.
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