Figure 4.
Figure 4. Soluble levels of IL-12, TNF-α, IL-10, and IL-6 from purified lung DCs obtained on day 15 after sham or CLP surgery. (A) Mice were subjected to sham or CLP surgeries and killed on day 15, and whole lung samples were processed. Lung DCs were purified by positive selection with CD11c beads. Purified DCs were plated at a density of 2 × 105/200 μL/well overnight, and then stimulated with medium, LPS (1 μg/mL), Pam3cys (25 μg/mL), SEA (100 μg/mL), or Aspergillus antigen (50 μg/mL) for 48 hours. Cell-free supernatants were collected, and IL-12, TNF-α, and IL-10 levels were measured by ELISA. (B) DCs were isolated and purified as described in panel A, and then cultured lung DCs were stimulated with medium, LPS (1 μg/mL), the legend regarding SEA (100 μg/mL), Aspergillus antigen (50 μg/mL), and Aspergillus conidia (1:1 ratio) and poli(I:C) (100 μg/mL) for 48 hours. Cell-free supernatants were collected and IL-6 level was measured by ELISA. Data are expressed as mean ± SEM. *P < .05 compared with the medium-treated group; #P < .05 compared with cytokine levels detected in cell-free supernatants from sham surgery lung DCs.

Soluble levels of IL-12, TNF-α, IL-10, and IL-6 from purified lung DCs obtained on day 15 after sham or CLP surgery. (A) Mice were subjected to sham or CLP surgeries and killed on day 15, and whole lung samples were processed. Lung DCs were purified by positive selection with CD11c beads. Purified DCs were plated at a density of 2 × 105/200 μL/well overnight, and then stimulated with medium, LPS (1 μg/mL), Pam3cys (25 μg/mL), SEA (100 μg/mL), or Aspergillus antigen (50 μg/mL) for 48 hours. Cell-free supernatants were collected, and IL-12, TNF-α, and IL-10 levels were measured by ELISA. (B) DCs were isolated and purified as described in panel A, and then cultured lung DCs were stimulated with medium, LPS (1 μg/mL), the legend regarding SEA (100 μg/mL), Aspergillus antigen (50 μg/mL), and Aspergillus conidia (1:1 ratio) and poli(I:C) (100 μg/mL) for 48 hours. Cell-free supernatants were collected and IL-6 level was measured by ELISA. Data are expressed as mean ± SEM. *P < .05 compared with the medium-treated group; #P < .05 compared with cytokine levels detected in cell-free supernatants from sham surgery lung DCs.

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