Fig. 7.
Fig. 7. Effect of IL-4, IL-10, IL-13, IL-6, IL-2, and LPS. / (A) Effect of IL-4, IL-10, IL-13, IL-6, IL-2, and LPS on A23187-stimulated production of LTB4. PMNs were incubated with IL-4 (10 ng/mL), IL-10 (100 ng/mL), IL-13 (100 ng/mL), IL-6 (100 ng/mL), IL-2 (10 ng/mL), or LPS (100 ng/mL) for 6 hours. A23187-stimulated LTB4 production was measured by RIA as described in “Materials and methods.” Data represent mean ± SEM (n = 4). *P < .05 versus control. (B) Effect of IL-4, IL-10, IL-13, IL-6, IL-2, and LPS on mRNA expression for LTA4 hydrolase. After PMNs were incubated with IL-4 (10 ng/mL), IL-10 (100 ng/mL), IL-13 (100 ng/mL), IL-6 (100 ng/mL), IL-2 (10 ng/mL), or LPS (100 ng/mL) for 6 hours, total RNA was isolated and RT-PCR for LTA4 hydrolase was performed as described in “Materials and methods.” (C) Northern blot analysis for LTA4 hydrolase. After PMNs were incubated with IL-4 (10 ng/mL), IL-10 (100 ng/mL), IL-13 (100 ng/mL), or LPS (100 ng/mL) for 6 hours, total RNA was isolated and Northern blot analysis for LTA4 hydrolase was performed as described in “Materials and methods.” The same membrane was rehybridized with a human beta-actin probe as a control. The positions of marker RNA are indicated by arrows. (D) Effect of IL-4, IL-10, IL-13, IL-6, IL-2, and LPS on protein expression of LTA4 hydrolase. PMNs were incubated with IL-4 (10 ng/mL), IL-10 (100 ng/mL), IL-13 (100 ng/mL), IL-6 (100 ng/mL), IL-2 (10 ng/mL), or LPS (100 ng/mL) for 6 hours. The cell lysates were prepared. Equal amount of protein (20 μg) were applied on 8% SDS-PAGE and electro-transferred onto nitrocellulose membrane. Western blot analysis was performed with anti-cPLA2, anti-5-LO, or anti-LTA4 hydrolase antibody as described in “Materials and methods.” The positions of molecular marker proteins are indicated by arrows. (E) Effect of IL-4, IL-10, IL-13, IL-6, IL-2, and LPS on LTA4 hydrolase activity. After PMNs were incubated with IL-4 (10 ng/mL), IL-10 (100 ng/mL), IL-13 (100 ng/mL), IL-6 (100 ng/mL), IL-2 (10 ng/mL), or LPS (100 ng/mL) for 6 hours, the cell lysates were prepared and incubated with LTA4 at 37°C for 5 minutes. The supernatant was recovered, and the concentration of LTB4 was measured by HPLC as described in “Materials and methods.” Data represent mean ± SEM (n = 4). *P < .05 versus control.

Effect of IL-4, IL-10, IL-13, IL-6, IL-2, and LPS.

(A) Effect of IL-4, IL-10, IL-13, IL-6, IL-2, and LPS on A23187-stimulated production of LTB4. PMNs were incubated with IL-4 (10 ng/mL), IL-10 (100 ng/mL), IL-13 (100 ng/mL), IL-6 (100 ng/mL), IL-2 (10 ng/mL), or LPS (100 ng/mL) for 6 hours. A23187-stimulated LTB4 production was measured by RIA as described in “Materials and methods.” Data represent mean ± SEM (n = 4). *P < .05 versus control. (B) Effect of IL-4, IL-10, IL-13, IL-6, IL-2, and LPS on mRNA expression for LTA4 hydrolase. After PMNs were incubated with IL-4 (10 ng/mL), IL-10 (100 ng/mL), IL-13 (100 ng/mL), IL-6 (100 ng/mL), IL-2 (10 ng/mL), or LPS (100 ng/mL) for 6 hours, total RNA was isolated and RT-PCR for LTA4 hydrolase was performed as described in “Materials and methods.” (C) Northern blot analysis for LTA4 hydrolase. After PMNs were incubated with IL-4 (10 ng/mL), IL-10 (100 ng/mL), IL-13 (100 ng/mL), or LPS (100 ng/mL) for 6 hours, total RNA was isolated and Northern blot analysis for LTA4 hydrolase was performed as described in “Materials and methods.” The same membrane was rehybridized with a human beta-actin probe as a control. The positions of marker RNA are indicated by arrows. (D) Effect of IL-4, IL-10, IL-13, IL-6, IL-2, and LPS on protein expression of LTA4 hydrolase. PMNs were incubated with IL-4 (10 ng/mL), IL-10 (100 ng/mL), IL-13 (100 ng/mL), IL-6 (100 ng/mL), IL-2 (10 ng/mL), or LPS (100 ng/mL) for 6 hours. The cell lysates were prepared. Equal amount of protein (20 μg) were applied on 8% SDS-PAGE and electro-transferred onto nitrocellulose membrane. Western blot analysis was performed with anti-cPLA2, anti-5-LO, or anti-LTA4 hydrolase antibody as described in “Materials and methods.” The positions of molecular marker proteins are indicated by arrows. (E) Effect of IL-4, IL-10, IL-13, IL-6, IL-2, and LPS on LTA4 hydrolase activity. After PMNs were incubated with IL-4 (10 ng/mL), IL-10 (100 ng/mL), IL-13 (100 ng/mL), IL-6 (100 ng/mL), IL-2 (10 ng/mL), or LPS (100 ng/mL) for 6 hours, the cell lysates were prepared and incubated with LTA4 at 37°C for 5 minutes. The supernatant was recovered, and the concentration of LTB4 was measured by HPLC as described in “Materials and methods.” Data represent mean ± SEM (n = 4). *P < .05 versus control.

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