1) To study the determinants of monocyte and macrophage host defenses against infection by the novel bacterial pathogen, Granulibacter bethesdensis Our recent publication (Zarember et al., Infection and Immunity, 2012) demonstrated that killing of G. bethesdensis by neutrophils from patients with chronic granulomatous disease (CGD) was defective and that this Gram-negative pathogen was remarkably resistant to complement and to cationic antimicrobial peptides. During FY13, Jessica Chu (Postdoctoral IRTA) completed a study demonstrating that CGD human monocytes and monocyte-derived macrophages were unable to kill G. bethesdensis normally and that this organism could persist in CGD macrophages. This paper also demonstrated a positive correlation between microbial killing by IFNgamma-treated monocytes and residual neutrophil NADPH oxidase activity. This finding has potential diagnostic implications in the use of IFN (Actimune) to treat CGD patients. 2) G. bethesdensis Lipopolysaccharide (LPS) During our studies of the interaction of Granulibacter bethesdensis with immune cells, we found that this organism is remarkably hypostimulatory of the human innate immune system, both in terms of weak activation of the NADPH oxidase and poor stimulation of cytokine secretion. We are collaborating with Yossi Shiloach (NIDDK) and Russell Carlson of the University of Georgia Complex Carbohydrate Research Center to complete the purification and structural characterization of the atypical lipopolysaccharide (LPS) of this organism and determine whether it acts as an anti-inflammatory inhibitory LPS. 3) G.bethesdensis Methanol Dehydrogenase In order to develop our serological testing for G.bethesdensis infection (see ZIA AI000155-36), we purified Methanol Dehydrogenase from this organism. Using ion exchange and gel filtration, highly enriched enzyme was prepared and biochemical testing is underway to identify substrate specificity, identify inhibitors, and s indicate a much broader substrate specificity that originally thought. In collaboration with Peter Steinbach (NIH Center for Molecular Modeling), we have modeled the structure of G. bethesdensis MDH.

Project Start
Project End
Budget Start
Budget End
Support Year
3
Fiscal Year
2013
Total Cost
$113,716
Indirect Cost
City
State
Country
Zip Code
Kuhns, Douglas B; Fink, Danielle L; Choi, Uimook et al. (2016) Cytoskeletal abnormalities and neutrophil dysfunction in WDR1 deficiency. Blood 128:2135-2143
De Ravin, Suk See; Wu, Xiaolin; Moir, Susan et al. (2016) Lentiviral hematopoietic stem cell gene therapy for X-linked severe combined immunodeficiency. Sci Transl Med 8:335ra57
De Ravin, Suk See; Reik, Andreas; Liu, Pei-Qi et al. (2016) Targeted gene addition in human CD34(+) hematopoietic cells for correction of X-linked chronic granulomatous disease. Nat Biotechnol 34:424-9
Feingold, Paul L; Quadri, Humair S; Steinberg, Seth M et al. (2016) Thoracic Surgery in Chronic Granulomatous Disease: a 25-Year Single-Institution Experience. J Clin Immunol 36:677-83
Kuhns, Douglas B; Long Priel, Debra A; Chu, Jessica et al. (2015) Isolation and Functional Analysis of Human Neutrophils. Curr Protoc Immunol 111:7.23.1-16
Greenberg, David E; Sturdevant, Daniel E; Marshall-Batty, Kimberly R et al. (2015) Simultaneous Host-Pathogen transcriptome analysis during Granulibacter bethesdensis infection of normal and chronic granulomatous disease neutrophils. Infect Immun :
Merling, Randall K; Sweeney, Colin L; Chu, Jessica et al. (2015) An AAVS1-targeted minigene platform for correction of iPSCs from all five types of chronic granulomatous disease. Mol Ther 23:147-57
Sibley, Christopher T; Estwick, Tyra; Zavodni, Anna et al. (2014) Assessment of atherosclerosis in chronic granulomatous disease. Circulation 130:2031-9
Chu, Jessica; Song, Helen H; Zarember, Kol A et al. (2013) Persistence of the bacterial pathogen Granulibacter bethesdensis in chronic granulomatous disease monocytes and macrophages lacking a functional NADPH oxidase. J Immunol 191:3297-307
Hsu, Amy P; Sowerwine, Kathryn J; Lawrence, Monica G et al. (2013) Intermediate phenotypes in patients with autosomal dominant hyper-IgE syndrome caused by somatic mosaicism. J Allergy Clin Immunol 131:1586-93

Showing the most recent 10 out of 14 publications