American Society of Hematology

Fig. 4.

$Fig. 4. Analysis of IL-8 in supernatant of PMNs or MNCs by ELISA. / Cells (4 × 105) were incubated in the presence of LPS, Gp96, or flanking fractions of the Gp96 purification scheme. Culture supernatants were harvested after 6 hours at 37°C with 7.5% CO2 and assayed for IL-8 by ELISA. To evaluate the contribution of cells other than PMNs to the release of IL-8, supernatants from PMNs (▪), PMNs with 5% MNCs added (■) (A) or MNCs only (B) were analyzed. All samples were taken in duplicates. The results shown are representative of 3 independent experiments. Statistical analysis of all 3 experiments indicated a significant increase in the release of IL-8 in the presence of Gp96 (at 100 μg/mL or 20 μg/mL for PMNs, P < .001; for PMNs+MNC or MNCs alone, P < .05) or LPS (at 10 ng/mL for PMNs,P = .0017; for PMNs+MNCs or MNCs, P < .05; flanking fraction+LPS, 10 ng/mL for PMNs, P < .001; for MNCs, P < .05), but not for flanking fraction alone (by Student t test).$

Analysis of IL-8 in supernatant of PMNs or MNCs by ELISA.

Cells (4 × 10⁵) were incubated in the presence of LPS, Gp96, or flanking fractions of the Gp96 purification scheme. Culture supernatants were harvested after 6 hours at 37°C with 7.5% CO₂ and assayed for IL-8 by ELISA. To evaluate the contribution of cells other than PMNs to the release of IL-8, supernatants from PMNs (▪), PMNs with 5% MNCs added (■) (A) or MNCs only (B) were analyzed. All samples were taken in duplicates. The results shown are representative of 3 independent experiments. Statistical analysis of all 3 experiments indicated a significant increase in the release of IL-8 in the presence of Gp96 (at 100 μg/mL or 20 μg/mL for PMNs, P < .001; for PMNs+MNC or MNCs alone, P < .05) or LPS (at 10 ng/mL for PMNs,P = .0017; for PMNs+MNCs or MNCs, P < .05; flanking fraction+LPS, 10 ng/mL for PMNs, P < .001; for MNCs, P < .05), but not for flanking fraction alone (by Student t test).

This Feature Is Available To Subscribers Only