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

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 6

Analysis of short-duration neutrophil–vessel wall interactions in control and anti-GBM–treated LysM-GFP mice.

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Analysis of short-duration neutrophil–vessel wall interactions in contro...
In vivo 2-photon video-rate imaging of glomeruli in untreated LysM-GFP mice (Ctrl) (n = 3 mice, 5 glomeruli) and in LysM-GFP mice 1.5–2 hours after anti-GBM (αGBM) treatment (n = 3 mice, 4 glomeruli). (A) A video-rate image of steady-state neutrophil (green) trafficking behavior in glomerular capillaries (red, Q-dot 565) (white arrows) of a control mouse (left panel) and a mouse after αGBM treatment (right panel). Images are 20 seconds of video-rate frames overlaid to show neutrophil tracks (green) in glomerular capillaries (red, Q-dot 565). Scale bars: 25 μm. (B) Distributions of cell dwell times in a single plane from video-rate recordings of control and αGBM mice plotted in seconds. Duration times were significantly different between the groups using a 2-tailed Mann-Whitney U test, P < 0.001. (C) Distributions using a 2-sample KS test comparison percentile plot. Group dwell time distributions were significantly different, P < 0.001, D = 0.6970. (D and E) Neutrophil velocities were plotted over time and fit to a hidden Markov 2-state model (HMM 2-state) to identify cell track transitions from fast moving, state 1 (S1), to adherent, state 2 (S2), behavior. Two representative cell tracks (1 and 2) from the control group (D) with typical steady-state behavior showed fluctuations in cell velocity, but no S1 to S2 transition, while those from the αGBM group (E) made brief S2 transitions (left panel) and sustained S2 transitions (right panel). (F) Mean dwell time ± SEM in seconds from video-rate recordings in control and αGBM groups. (G) Average S1 to S2 transition frequency per track in control and αGBM groups. S2 transitions were more frequent in the αGBM group. (H) Average S2 durations were more prolonged in the αGBM compared with the control group. (I) Average neutrophil S1 velocities (μm/s) were higher in the control versus the αGBM group. *P < 0.01, **P < 0.001, by 2-tailed unpaired t test for F–I.