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 7

Rapid neutrophil capture in glomerular capillaries is FcγRIIA and Abl/Src dependent.

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Rapid neutrophil capture in glomerular capillaries is FcγRIIA and Abl/Sr...
(A–F) FcγRIIA+/γ–/– recipient mice were given anti-GBM antibody (αGBM) (A–C) or left untreated (D–F), and 2-photon time-lapse imaging of the kidney was conducted 2 hours later following the simultaneous adoptive transfer of FcγRIIA+/γ–/– (IIAγ–/–) and γ–/– neutrophils differentially labeled with green or blue fluorescent dye and a red dextran vascular dye. Duration times were calculated as the time in seconds that neutrophils remained arrested in the capillary and the distribution of dwell times plotted as histograms. (A–C) In αGBM-treated recipients given IIAγ–/– and γ–/– neutrophils, the group and dwell time distributions were compared using a Mann-Whitney U test (A, P < 0.001), a 2-sample KS test comparison percentile plot (B, P < 0.001, D = 0.2485), and a 2-tailed unpaired t test (C, *P < 0.001) (n = 4 recipient mice with n = 2 for each neutrophil dye combination, 33 glomeruli). (D–F) Untreated recipients were analyzed following the adoptive transfer of labeled IIAγ–/– and γ–/– neutrophils, and statistics were conducted as in A–C (D, P = 0.138; E, P = 0.138, D = 0.1859; F, P = 0.138) (n = 3 recipient mice with 1 dye combination, 21 glomeruli). (G–I) FcγRIIA+/γ–/– recipient mice were given αGBM, and time-lapse imaging was conducted 2 hours after the adoptive transfer of differentially labeled FcγRIIA+/γ–/– neutrophils pretreated with 100 nM bosutinib (Bos) or vehicle (DMSO). Duration time (seconds) and group dwell time distributions were compared as in A–C. Statistics are as follows: G, P < 0.001; H, P < 0.001 and D = 0.2346; I, *P < 0.001 (n = 4 recipient mice with n = 2 for each neutrophil dye combination, 22 glomeruli). Bar graphs in C, F, and I are mean duration time ± SEM in seconds.