Chronic granulomatous disease (CGD) is a rare, inherited disorder of various defects of the NADPH oxidase rendering it dysfunctional and predisposing to invasive and life-threatening bacterial and fungal infection. While the role of the defective oxidase in immunodeficiency is undisputed, disordered inflammation in the disease (e.g., poor wound healing, obstructing granulomata and autoimmunity), which may or may not be associated with infection, remains unexplained. It is hypothesized that disordered inflammation in CGD results from defective phosphatidylserine (PS) exposure and accumulation on apoptosing CGD neutrophils that leads to impaired recognition and engulfment by phagocytes. Preliminary data suggest that the functioning NADPH oxidase is required for appropriate PS exposure during neutrophil apoptosis, an event that requires coordinate activation of phospholipid (PL) scrambling to expose PS, and inactivation of the aminophospholipid translocase which returns PS to the plasma membrane inner leaflet. This proposal seeks to define the defect in PS exposure on CGD neutrophils by assessing both PL scrambling and aminophospholipid translocase activity. It is hypothesized that apoptosing CGD neutrophils will have diminished PL scrambling and sustained aminophospholipid translocase activity. Exposure of PS on apoptotic cells is required for recognition by phagocytes and is mediated through a receptor, the PSR, that recognizes PS in a stereospeciflc manner. Engagement of the PSR along with other tethering receptors mediates the ingestion of apoptotic neutrophils by an immunologically """"""""silent"""""""" mechanism that results in TGFbeta production and suppression of pro-inflammatory cytokine production from the phagocyte. Thus, we hypothesized that PS deficient apoptotic CGD neutrophils will show impaired phagocytic recognition and engulfment, and promote pro-inflammatory cytokine production. Furthermore, unengulfed CGD neutrophils will undergo post-apoptotic cytolysls releasing injurious proteins. Both in vitro phagocytic engulfment assays and in vivo murine models are proposed. It is predicted that apoptotic CGD neutrophils, relative to apoptotic normal neutrophils, instilled into the inflamed peritonea of wild type mice will show delayed clearance, suppression of TGFbeta production and enhanced pro-inflammatory cytokine production; restoration of PS on the surface of CGD neutrophils will reverse this. Finally, injection of CGD mice with sterile fungal hyphae in the presence of PS liposomes will lead to enhanced clearance of accumulating neutrophils, enhanced TGFbeta production and resolution of inflammation. It is expected that data from this investigation will identify novel potential therapeutic targets to address disordered inflammation of CGD.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
1R01AI058228-01
Application #
6720011
Study Section
Erythrocyte and Leukocyte Biology Study Section (ELB)
Program Officer
Plaut, Marshall
Project Start
2003-12-15
Project End
2007-11-30
Budget Start
2003-12-15
Budget End
2004-11-30
Support Year
1
Fiscal Year
2004
Total Cost
$265,300
Indirect Cost
Name
National Jewish Health
Department
Type
DUNS #
076443019
City
Denver
State
CO
Country
United States
Zip Code
80206
Fernandez-Boyanapalli, Ruby F; Falcone, Emilia Liana; Zerbe, Christa S et al. (2015) Impaired efferocytosis in human chronic granulomatous disease is reversed by pioglitazone treatment. J Allergy Clin Immunol 136:1399-1401.e3
Fernandez-Boyanapalli, Ruby F; Frasch, S Courtney; Thomas, Stacey M et al. (2015) Pioglitazone restores phagocyte mitochondrial oxidants and bactericidal capacity in chronic granulomatous disease. J Allergy Clin Immunol 135:517-527.e12
Redente, Elizabeth F; Keith, Rebecca C; Janssen, William et al. (2014) Tumor necrosis factor-? accelerates the resolution of established pulmonary fibrosis in mice by targeting profibrotic lung macrophages. Am J Respir Cell Mol Biol 50:825-37
Henson, Peter M; Bratton, Donna L (2013) Antiinflammatory effects of apoptotic cells. J Clin Invest 123:2773-4
Fernandez-Boyanapalli, Ruby; Goleva, Elena; Kolakowski, Christena et al. (2013) Obesity impairs apoptotic cell clearance in asthma. J Allergy Clin Immunol 131:1041-7, 1047.e1-3
Frasch, S Courtney; Fernandez-Boyanapalli, Ruby F; Berry, Karin A Zemski et al. (2013) Neutrophils regulate tissue Neutrophilia in inflammation via the oxidant-modified lipid lysophosphatidylserine. J Biol Chem 288:4583-93
Xiong, Weipeng; Frasch, S Courtney; Thomas, Stacey M et al. (2013) Induction of TGF-?1 synthesis by macrophages in response to apoptotic cells requires activation of the scavenger receptor CD36. PLoS One 8:e72772
Frasch, S Courtney; Bratton, Donna L (2012) Emerging roles for lysophosphatidylserine in resolution of inflammation. Prog Lipid Res 51:199-207
Frasch, S Courtney; Fernandez-Boyanapalli, Ruby F; Berry, Karin Zemski et al. (2011) Signaling via macrophage G2A enhances efferocytosis of dying neutrophils by augmentation of Rac activity. J Biol Chem 286:12108-22
Kenyon, Karla D; Cole, Caroline; Crawford, Fran et al. (2011) IgG autoantibodies against deposited C3 inhibit macrophage-mediated apoptotic cell engulfment in systemic autoimmunity. J Immunol 187:2101-11

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