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 2

Bosutinib inhibits FcγRIIA functions in human neutrophils.

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Bosutinib inhibits FcγRIIA functions in human neutrophils. (A) ROS after...
(A) ROS after FcγRIIA cross-linking (XL) (left) or PMA (right) in bosutinib-pretreated (Bos) and PP2-pretreated neutrophils as in Figure 1, C and D (n = 3). (B) Representative profile of XL-induced ROS after treatment with vehicle or 100 nM Bos, imatinib (Ima), or nilotinib (Nil) (n = 3). (C) Neutrophils pretreated with 100 nM of the indicated compounds, 1 μM PP2, or vehicle (DMSO, –) on BSA- or IC-coated plates, stained with fluorescent phalloidin, and quantitated for adherent cells (left) and spread cell area (right) (n = 3). Representative images are shown. Original magnification, ×600. (D) The number of adherent cells pretreated as in C and drawn across BSA- or IC-coated plates at 1.0 dyn/cm2 was calculated and is presented as the fold induction compared with vehicle (–)/BSA control (n = 3, except for Nil, n = 2). (E) HDMECs were treated without (–) or with (+) TNF and/or anti-CD105 plus a secondary antibody (2°ab). Neutrophils pretreated with anti-FcγRIIIB (3G8), FcγRIIA (IV.3), or isotype (Iso) controls (left panel), or with 100 nM Bos or Ima (right panel), were drawn across HDMECs at 1.5 dyn/cm2. The number of rolling and adherent cells was assessed. Fold induction was relative to untreated HDMECs (n = 3). (F) XL- or PMA-induced ROS in blood leukocytes from 3 CML patients receiving Bos and matched normal volunteers (Ctrl). A representative ROS profile (left panel) and average fold induction relative to the XL (Ctrl) sample (right panel) are shown (n = 3). (G) CML and control leukocytes were analyzed as in D at 1.0 and 1.5 dyn/cm2. Fold induction is relative to the control sample on BSA (Ctrl/BSA) (n = 3). All data are expressed as the mean ± SEM. *P < 0.05, **P < 0.01, 1-way ANOVA followed by Dunnett’s multiple comparisons test for A to assess dose responsiveness of the drugs compared with vehicle and also for C–E and G, and by 2-tailed unpaired t test in F.