Neutrophil FcγRIIA promotes IgG-mediated glomerular neutrophil capture via Abl/Src kinases
Hiroshi Nishi, … , George C. Tsokos, Tanya N. Mayadas
Hiroshi Nishi, … , George C. Tsokos, Tanya N. Mayadas
Published September 11, 2017
Citation Information: J Clin Invest. 2017;127(10):3810-3826. https://doi.org/10.1172/JCI94039.
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Research Article Inflammation Nephrology Article has an altmetric score of 5

Neutrophil FcγRIIA promotes IgG-mediated glomerular neutrophil capture via Abl/Src kinases

Abstract

The kidney glomerular capillaries are frequent sites of immune complex deposition and subsequent neutrophil accumulation in post-infectious and rapidly progressive glomerulonephritis. However, the mechanisms of neutrophil recruitment remain enigmatic, and there is no targeted therapeutic to avert this proximal event in glomerular inflammation. The uniquely human activating Fc receptor FcγRIIA promotes glomerular neutrophil accumulation and damage in anti–glomerular basement membrane–induced (anti-GBM–induced) glomerulonephritis when expressed on murine neutrophils. Here, we found that neutrophils are directly captured by immobilized IgG antibodies under physiological flow conditions in vitro through FcγRIIA-dependent, Abl/Src tyrosine kinase–mediated F-actin polymerization. Biophysical measurements showed that the lifetime of FcγRIIA-IgG bonds increased under mechanical force in an F-actin–dependent manner, which could enable the capture of neutrophils under physiological flow. Kidney intravital microscopy revealed that circulating neutrophils, which were similar in diameter to glomerular capillaries, abruptly arrested following anti-GBM antibody deposition via neutrophil FcγRIIA and Abl/Src kinases. Accordingly, inhibition of Abl/Src with bosutinib reduced FcγRIIA-mediated glomerular neutrophil accumulation and renal injury in experimental, crescentic anti-GBM nephritis. These data identify a pathway of neutrophil recruitment within glomerular capillaries following IgG deposition that may be targeted by bosutinib to avert glomerular injury.

Authors

Hiroshi Nishi, Kazuhiro Furuhashi, Xavier Cullere, Gurpanna Saggu, Mark J. Miller, Yunfeng Chen, Florencia Rosetti, Samantha L. Hamilton, Lihua Yang, Spencer P. Pittman, Jiexi Liao, Jan M. Herter, Jeffrey C. Berry, Daniel J. DeAngelo, Cheng Zhu, George C. Tsokos, Tanya N. Mayadas

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Figure 4

Actin polymerization is required for force-induced FcγRIIA-mediated interaction with IgG.

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Actin polymerization is required for force-induced FcγRIIA-mediated inte...
(A) Human peripheral blood neutrophils were treated with 2 different doses of cytochalasin D (cytD) or with DMSO (–), and adhesion of cells to an IC- or BSA-coated surface under flow (1.0 dyn/cm2) was assessed. Fold induction is relative to vehicle-treated (–) samples on ICs (n = 3). (B) Human peripheral blood neutrophils were pretreated with cytD and drawn across HDMECs treated with TNF and anti-endoglin as indicated and detailed in Figure 2E. Fold induction is relative to untreated HDMECs (n = 3). (C) Human neutrophils were treated with 2 different doses of cytD or vehicle (DMSO, –), and the number of cells adherent to IC- or BSA-coated surfaces under static conditions was assessed (n = 3). (D and E) Jurkat cells expressing FcγRIIA but mutagenized to lack CD18 integrins were evaluated for adhesion to ICs or BSA under flow (D) or static (E) conditions in the presence or absence of cytD (n = 3). Representative images for the static assay are shown to illustrate changes in actin distribution in cytD-treated versus vehicle-treated (–) neutrophils adherent to ICs. Scale bar: 25 μm. (F) Force-dependent lifetimes of single bonds (in seconds) between hIgG1-immobilized beads and FcγRIIA-transfected Jurkat cells in the presence (blue circles) or absence (–, red squares) of cytD. Each point represents more than 30 repeated measurements. Data are expressed as mean ± SEM. *P < 0.05, **P < 0.01, 1-way ANOVA followed by Dunnett’s multiple comparison test for A–E and 2-tailed unpaired t test for F.