During FY16, we continued our studies of Granulibacter bethesdensis, an emerging pathogen in patients with chronic granulomatous disease (CGD). We completed our comparative analysis of the transcriptomes of Granulibacter exposed to normal or CGD PMN and, in parallel, the transcriptomes of normal and CGD PMN following exposure to Granulibacter (Greenberg et al., 2015). This study documented possible mechanisms by which Granulibacter delays neutrophil apoptosis, and identified ClpB, a stress response gene in Granulibacter, as a virulence factor required for survival in PMN. Alterations in Granulibacter metabolic gene expression during intracellular growth lead to the identification of pyruvate dehydrogenase as an essential gene and that a pharmacologic inhibitor could be used as an antibiotic against Granulibacter in vitro. During FY16, we have significantly advanced our study of the genomes and phenotypic attributes of individual patient Granulibacter isolates (9 genetically distinct isolates) provided by NIH and international collaborators in Spain and in Portugal. Total genome sequencing of these isolates was performed and is being analyzed in combination with established laboratory assays of immune cell function (e.g., phagocytosis and killing by neutrophils, monocytes, and macrophages). Given our previous finding that G. bethesdensis persists in macrophages in vitro, we examined the intracellular trafficking of this organism to help explain its intracellular growth. This work is currently in preparation for publication. In collaboration with Artur Muszyski of the University of Georgia Complex Carbohydrate Research Center, NMR studies of the lipid A Granulibacter bethesdensis are underway to determine whether the chemical nature of this material accounts for the relatively poor ability of Granulibacter to activate human immune cells such as PMN and monocytes.

Project Start
Project End
Budget Start
Budget End
Support Year
6
Fiscal Year
2016
Total Cost
Indirect Cost
Name
Niaid Extramural Activities
Department
Type
DUNS #
City
State
Country
Zip Code
Myles, Ian A; Anderson, Erik D; Earland, Noah J et al. (2018) TNF overproduction impairs epithelial staphylococcal response in hyper IgE syndrome. J Clin Invest 128:3595-3604
Chu, Jessica; Smelkinson, Margery G; Dorward, David W et al. (2017) Early Intracellular Trafficking of Granulibacter bethesdensis in Human Macrophages. Infect Immun 85:
De Ravin, Suk See; Li, Linhong; Wu, Xiaolin et al. (2017) CRISPR-Cas9 gene repair of hematopoietic stem cells from patients with X-linked chronic granulomatous disease. Sci Transl Med 9:
Kuhns, Douglas B; Fink, Danielle L; Choi, Uimook et al. (2016) Cytoskeletal abnormalities and neutrophil dysfunction in WDR1 deficiency. Blood 128:2135-2143
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
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
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
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 :
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

Showing the most recent 10 out of 18 publications