Figure 4.
Intestinal IL-33–ST2 signaling promotes neutrophil recruitment in acute peritonitis via platelet-derived 5-HT. (A-F) Intraperitoneal injection of 4% thioglycollate into Il33fl/fl and Vil1creIl33fl/fl (A-C) or Il1rl1fl/fl and ChgacreERIl1rl1fl/fl (D-F) mice. (A,D) Flow cytometric analysis of lymphocytes isolated from peritoneum 4 hours after thioglycollate administration. (B,E) Quantification of percentile of gated F4/80–Ly6G+ neutrophils in panels A and D. (D,F) Quantification of F4/80−Ly6G+ neutrophils isolated from peritoneum 4 hours after thioglycollate administration. (G-I) Tph1fl/fl and Vil1creTph1fl/fl mice were treated with PBS or IL-33 followed by a 10-minute wait. Intraperitoneal injection of 4% thioglycollate into Tph1fl/fl and Vil1creTph1fl/fl mice. (G) Flow cytometric analysis of lymphocytes isolated from peritoneum 4 hours after thioglycollate administration with or without IL-33 treatment. (H) Quantification of percentile of gated F4/80−Ly6G+ neutrophils in panel G. (I) Quantification of F4/80−Ly6G+ neutrophils isolated from peritoneum 4 hours after thioglycollate administration with or without IL-33 treatment. (J) Schematic illustration of platelet depletion in WT mice, followed by intraperitoneal injection of 4% thioglycolate with or without IL-33 injection. (K) Flow cytometric analysis of lymphocytes isolated from peritoneum as in panel J. (L) Quantification of percentile of gated F4/80−Ly6G+ neutrophils in panel K. (M) Quantification of F4/80−Ly6G+ neutrophils isolated from peritoneum 4 hours after peritonitis induction. (N) Schematic illustration of fluoxetine treatment in WT mice, followed by intraperitoneal injection of 4% thioglycolate. (O) Flow cytometric analysis of lymphocytes isolated from peritoneum as in panel N. (P) Quantification of percentile of gated F4/80−Ly6G+ neutrophils in panel O. (Q) Quantification of F4/80−Ly6G+ neutrophils isolated from peritoneum 4 hours after peritonitis induction with or without 10-minute IL-33 treatment. Data are representative of 2 independent experiments (A, D, G, K, O) or are pooled from 2 independent experiments (B, C, E-F, H-I, L-M, P-Q). *P < .05, **P < .01, ***P < .001 by Student t test. Error bars represent standard deviation. NS, not statistically significant.

Intestinal IL-33–ST2 signaling promotes neutrophil recruitment in acute peritonitis via platelet-derived 5-HT. (A-F) Intraperitoneal injection of 4% thioglycollate into Il33fl/fl and Vil1creIl33fl/fl (A-C) or Il1rl1fl/fl and ChgacreERIl1rl1fl/fl (D-F) mice. (A,D) Flow cytometric analysis of lymphocytes isolated from peritoneum 4 hours after thioglycollate administration. (B,E) Quantification of percentile of gated F4/80–Ly6G+ neutrophils in panels A and D. (D,F) Quantification of F4/80−Ly6G+ neutrophils isolated from peritoneum 4 hours after thioglycollate administration. (G-I) Tph1fl/fl and Vil1creTph1fl/fl mice were treated with PBS or IL-33 followed by a 10-minute wait. Intraperitoneal injection of 4% thioglycollate into Tph1fl/fl and Vil1creTph1fl/fl mice. (G) Flow cytometric analysis of lymphocytes isolated from peritoneum 4 hours after thioglycollate administration with or without IL-33 treatment. (H) Quantification of percentile of gated F4/80−Ly6G+ neutrophils in panel G. (I) Quantification of F4/80−Ly6G+ neutrophils isolated from peritoneum 4 hours after thioglycollate administration with or without IL-33 treatment. (J) Schematic illustration of platelet depletion in WT mice, followed by intraperitoneal injection of 4% thioglycolate with or without IL-33 injection. (K) Flow cytometric analysis of lymphocytes isolated from peritoneum as in panel J. (L) Quantification of percentile of gated F4/80−Ly6G+ neutrophils in panel K. (M) Quantification of F4/80−Ly6G+ neutrophils isolated from peritoneum 4 hours after peritonitis induction. (N) Schematic illustration of fluoxetine treatment in WT mice, followed by intraperitoneal injection of 4% thioglycolate. (O) Flow cytometric analysis of lymphocytes isolated from peritoneum as in panel N. (P) Quantification of percentile of gated F4/80−Ly6G+ neutrophils in panel O. (Q) Quantification of F4/80−Ly6G+ neutrophils isolated from peritoneum 4 hours after peritonitis induction with or without 10-minute IL-33 treatment. Data are representative of 2 independent experiments (A, D, G, K, O) or are pooled from 2 independent experiments (B, C, E-F, H-I, L-M, P-Q). *P < .05, **P < .01, ***P < .001 by Student t test. Error bars represent standard deviation. NS, not statistically significant.

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