Deposition of antibody-antigen immune complexes (IC) in tissues underlies the pathogenesis of a range of human autoimmune diseases from glomerulonephritis, SLE and arthritis, to transplantation rejection. Infiltrating leukocytes are the cause of inflammatory tissue damage. The primary objective of this competing renewal remains an in depth study of the mechanisms of IC mediated neutrophil recruitment and injury. Receptors for IgG, collectively known as Fc?Rs, mediate the interaction of antibodies with effector cells. In the current grant period, we provide several lines of evidence that the low affinity activating neutrophil Fc?Rs in mice, which preferentially complex with IgG-ICs, directly mediate neutrophil accumulation. Neutrophil Fc?Rs and the leukocyte specific CD18 integrins cooperated to promote IC induced neutrophil slow rolling, adhesion and transmigration, and TNF priming of neutrophils enhanced these processes. There are notable structural differences between neutrophil human and mouse low affinity activating Fc?Rs. Mice express Fc?RIII and Fc?RIV which complex with the common ?-chain to signal while human neutrophils express Fc?RIIA with the ITAM signaling and ligand binding domain in a single polypeptide chain, and a unique GPI-linked Fc?RIIIB. To examine the physiological functions, and the relative roles of the human neutrophil Fc?Rs in vivo, we expressed human Fc?RIIIB and Fc?RIIA selectively in neutrophils of mice lacking the common ?-chain (?-/-) and thus endogenous activating Fc?Rs. ?-/- mice failed to develop anti-glomerular basement membrane induced progressive glomerulonephritis and soluble IC induced Reverse Passive Arthus reaction as expected. In contrast, mice expressing hFc?RIIA on neutrophils displayed significant tissue injury. Moreover, mice expressing either hFc?RIIIB or hFc?RIIA exhibited significant tissue neutrophil accumulation in both models. Using intravital microscopy, we demonstrate that neutrophil Fc?RIIA and Fc?RIIIB play specialized context-dependent roles in IC-mediated neutrophil slow rolling, adhesion and transmigration. From these, and related observations, we hypothesize that Fc?RIIA and Fc?RIIIB serve distinct nonredundant functions in response to ICs, which are differentially regulated. We propose that 1) Fc?RIIIB tethering to ICs, supported by CD18 integrins serves a homeostatic role in clearing intravascular ICs, while 2) Fc?RIIA mediated recruitment and associated injury is regulated by cytokines, resident mast cells and macrophages, and requires its ITAM motif. In this proposal we will rigorously test these hypothesis using our humanized Fc?R transgenics, mouse lines deficient in mast cells or macrophages, and cell reconstitution approaches coupled with well-characterized in vivo models of IC induced inflammation and intravital microscopy. We anticipate that the new information provided by this proposal will increase our understanding of mechanisms of IC-induced neutrophil recruitment and activation and provide therapeutic targets for antibody-mediated inflammation relevant to human disorders.

Public Health Relevance

The objective of this proposal is to understand the role of human neutrophil Fc?Rs, receptors for IgG, in autoimmune diseases, which afflict 5-8% of the population. In mice, Fc?Rs have emerged as critical players in IC induced injury of the lung, skin and joints. However, the relevance of this to human inflammation remains unclear, as there are significant structural differences between activating Fc?Rs in mice and humans. We have generated mice expressing the two uniquely human activating neutrophil Fc?Rs, Fc?RIIA and Fc?RIIIB selectively in neutrophils and shown a predominant role for these human receptors in the initiation of immune complex-mediated diseases. Here we will further determine the physiological role of these receptors in neutrophil recruitment and injury in vivo. We anticipate that this will lead to the identification of therapeutic strategies that target tissue injury in inflammatory and autoimmune disorders relevant to human disease.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL065095-14
Application #
8402846
Study Section
Special Emphasis Panel (ZRG1-RUS-C (03))
Program Officer
Shah, Monica R
Project Start
1999-09-30
Project End
2013-12-10
Budget Start
2012-12-01
Budget End
2013-12-10
Support Year
14
Fiscal Year
2013
Total Cost
$416,883
Indirect Cost
$181,263
Name
Brigham and Women's Hospital
Department
Type
DUNS #
030811269
City
Boston
State
MA
Country
United States
Zip Code
02115
Mayadas, Tanya N; Cullere, Xavier; Lowell, Clifford A (2014) The multifaceted functions of neutrophils. Annu Rev Pathol 9:181-218
Venkatesh, Deepak; Ernandez, Thomas; Rosetti, Florencia et al. (2013) Endothelial TNF receptor 2 induces IRF1 transcription factor-dependent interferon-? autocrine signaling to promote monocyte recruitment. Immunity 38:1025-37
Niewczas, Monika A; Gohda, Tomohito; Skupien, Jan et al. (2012) Circulating TNF receptors 1 and 2 predict ESRD in type 2 diabetes. J Am Soc Nephrol 23:507-15
Gohda, Tomohito; Niewczas, Monika A; Ficociello, Linda H et al. (2012) Circulating TNF receptors 1 and 2 predict stage 3 CKD in type 1 diabetes. J Am Soc Nephrol 23:516-24
Tsuboi, Naotake; Ernandez, Thomas; Li, Xun et al. (2011) Regulation of human neutrophil Fc? receptor IIa by C5a receptor promotes inflammatory arthritis in mice. Arthritis Rheum 63:467-78
Li, Xun; Utomo, Ahmad; Cullere, Xavier et al. (2011) The ?-glucan receptor Dectin-1 activates the integrin Mac-1 in neutrophils via Vav protein signaling to promote Candida albicans clearance. Cell Host Microbe 10:603-15
Mayadas, Tanya N; Rosetti, Florencia; Ernandez, Thomas et al. (2010) Neutrophils: game changers in glomerulonephritis? Trends Mol Med 16:368-78
Hirahashi, Junichi; Hishikawa, Keiichi; Kaname, Shinya et al. (2009) Mac-1 (CD11b/CD18) links inflammation and thrombosis after glomerular injury. Circulation 120:1255-65
Mayadas, Tanya N; Tsokos, George C; Tsuboi, Naotake (2009) Mechanisms of immune complex-mediated neutrophil recruitment and tissue injury. Circulation 120:2012-24
Cullere, Xavier; Lauterbach, Michael; Tsuboi, Naotake et al. (2008) Neutrophil-selective CD18 silencing using RNA interference in vivo. Blood 111:3591-8

Showing the most recent 10 out of 19 publications