MST1-dependent vesicle trafficking regulates neutrophil transmigration through the vascular basement membrane
Angela R.M. Kurz, … , Sergio D. Catz, Markus Sperandio
Angela R.M. Kurz, … , Sergio D. Catz, Markus Sperandio
Published November 1, 2016; First published October 4, 2016
Citation Information: J Clin Invest. 2016;126(11):4125-4139. https://doi.org/10.1172/JCI87043.
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Categories: Research Article Inflammation

MST1-dependent vesicle trafficking regulates neutrophil transmigration through the vascular basement membrane

Abstract

Neutrophils need to penetrate the perivascular basement membrane for successful extravasation into inflamed tissue, but this process is incompletely understood. Recent findings have associated mammalian sterile 20–like kinase 1 (MST1) loss of function with a human primary immunodeficiency disorder, suggesting that MST1 may be involved in immune cell migration. Here, we have shown that MST1 is a critical regulator of neutrophil extravasation during inflammation. Mst1-deficient (Mst1–/–) neutrophils were unable to migrate into inflamed murine cremaster muscle venules, instead persisting between the endothelium and the basement membrane. Mst1–/– neutrophils also failed to extravasate from gastric submucosal vessels in a murine model of Helicobacter pylori infection. Mechanistically, we observed defective translocation of VLA-3, VLA-6, and neutrophil elastase from intracellular vesicles to the surface of Mst1–/– neutrophils, indicating that MST1 is required for this crucial step in neutrophil transmigration. Furthermore, we found that MST1 associates with the Rab27 effector protein synaptotagmin-like protein 1 (JFC1, encoded by Sytl1 in mice), but not Munc13-4, thereby regulating the trafficking of Rab27-positive vesicles to the cellular membrane. Together, these findings highlight a role for MST1 in vesicle trafficking and extravasation in neutrophils, providing an additional mechanistic explanation for the severe immune defect observed in patients with MST1 deficiency.

Authors

Angela R.M. Kurz, Monika Pruenster, Ina Rohwedder, Mahalakshmi Ramadass, Kerstin Schäfer, Ute Harrison, Gabriel Gouveia, Claudia Nussbaum, Roland Immler, Johannes R. Wiessner, Andreas Margraf, Dae-Sik Lim, Barbara Walzog, Steffen Dietzel, Markus Moser, Christoph Klein, Dietmar Vestweber, Rainer Haas, Sergio D. Catz, Markus Sperandio

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Figure 1

MST1 is dispensable for neutrophil adhesion in humans and mice, but critical for neutrophil extravasation into inflamed peritoneum.

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MST1 is dispensable for neutrophil adhesion in humans and mice, but crit...
(A) MST1 and MST2 protein levels of WT and Mst1–/– neutrophils. GAPDH served as loading control (n = 3). (B) Differential blood counts of WT (n = 20) and Mst1–/– mice (n = 14) (scatter plot with mean, ***P < 0.001, 2-way ANOVA, Sidak’s multiple comparisons test). (C) Neutrophil adhesion efficiency (arbitrary units) in cremaster muscle venules of WT and Mst1–/– mice, 2 hours after intrascrotal injection of TNF-α (n = 5, mean ± SEM, NS, unpaired t test). (D) Number of adherent leukocytes per field of view (FOV) in microflow chambers coated with E-selectin, E-selectin/ICAM-1, or E-selectin/ICAM-1/CXCL1 (n = 3, mean ± SEM, NS, 2-way ANOVA, Sidak’s multiple comparisons test). (E) LFA-1 clustering of WT or Mst1–/– cells labeled with anti–LFA-1 antibody conjugated with Alexa 546 (fluorescence intensity scale). Whole blood was perfused through microflow chambers coated with E-selectin, ICAM-1, and CXCL1 (Supplemental Videos 1 and 2) (n = 3 mice, 1 representative picture is shown). Scale bar: 10 μm. (F) Number of adherent neutrophils per field of view from 2 patients with STK4 deficiency and their heterozygous parents was calculated over time in flow chambers coated with E-selectin/ICAM-1/CXCL8 (n = 2, chambers ≥2, mean ± SEM). (G) Total number of extravasated neutrophils in the peritoneal lavage 2 hours after i.p. injection of NaCl or TNF-α in WT and Mst1–/– mice (n = 5, scatter plot with mean, ***P < 0.001, 2-way ANOVA, Tukey’s multiple comparisons test).