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. 2022 Nov 10;13:994496. doi: 10.3389/fimmu.2022.994496

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Copyright © 2022 Pérez-Figueroa, Álvarez-Carrasco and Ortega

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Production of inflammatory cytokines by Neutrophils stimulated by CD13 crosslinking. Neutrophils were incubated with Fab fragments of anti-CD13 mAbs (452 or C), Fab fragments of anti-FcγRII (Fab IV.3), or with anti- FcγRIIIb mAb (mAb CD16b) with or without secondary F(ab’)2 fragments of goat anti-mouse Ig, as indicated. As control, cells were stimulated with LPS (positive control) or with F(ab’)2 fragments alone. The cells were incubated for 24 hours and the cell culture supernatants were recovered. Concentrations of IL-1β (A), TNF-α (B) and IL-8 (C) in the supernatants were quantitated by a multiplex array. Results are expressed as mean ± SEM of 3 independent experiments using cells from 3 different donors. Statistical significance was calculated using one-way ANOVA with Tukey post hoc test (**p< 0.01, ***p< 0.001). Asterisks shown above each bar indicate statistical difference as compared to cytokine production by cells incubated with F(ab’)2 fragments of anti-mouse Ig alone (third bar in each graph). In (C), asterisks (**) above the line () drawn above the bar for Fab452 alone (penultimate bar) refer to the comparison between FabC plus secondary F(ab’)2 fragments and the Fab452 plus secondary F(ab’)2 fragments, that show a statistically significant difference between them. *p<0.05.

Inline graphic — Production of inflammatory cytokines by Neutrophils stimulated by CD13 crosslinking. Neutrophils were incubated with Fab fragments of anti-CD13 mAbs (452 or C), Fab fragments of anti-FcγRII (Fab IV.3), or with anti- FcγRIIIb mAb (mAb CD16b) with or without secondary F(ab’)2 fragments of goat anti-mouse Ig, as indicated. As control, cells were stimulated with LPS (positive control) or with F(ab’)2 fragments alone. The cells were incubated for 24 hours and the cell culture supernatants were recovered. Concentrations of IL-1β (A), TNF-α (B) and IL-8 (C) in the supernatants were quantitated by a multiplex array. Results are expressed as mean ± SEM of 3 independent experiments using cells from 3 different donors. Statistical significance was calculated using one-way ANOVA with Tukey post hoc test (**p< 0.01, ***p< 0.001). Asterisks shown above each bar indicate statistical difference as compared to cytokine production by cells incubated with F(ab’)2 fragments of anti-mouse Ig alone (third bar in each graph). In (C), asterisks (**) above the line () drawn above the bar for Fab452 alone (penultimate bar) refer to the comparison between FabC plus secondary F(ab’)2 fragments and the Fab452 plus secondary F(ab’)2 fragments, that show a statistically significant difference between them. *p<0.05.