Simultaneous Host-Pathogen Transcriptome Analysis during Granulibacter bethesdensis Infection of Neutrophils from Healthy Subjects and Patients with Chronic Granulomatous Disease.
Polymorphonuclear leukocytes (PMN) from patients with chronic granulomatous disease (CGD) fail to produce microbicidal concentrations of reactive oxygen species (ROS) due to mutations in NOX2. Patients with CGD suffer from severe, life-threatening infections and inflammatory complications. Granulibacter bethesdensis is an emerging Gram-negative pathogen in CGD that resists killing by PMN of CGD patients (CGD PMN) and inhibits PMN apoptosis through unknown mechanisms. Microarray analysis was used to study mRNA expression in PMN from healthy subjects (normal PMN) and CGD PMN during incubation with G. bethesdensis and, simultaneously, in G. bethesdensis with normal and CGD PMN. We detected upregulation of antiapoptotic genes (e.g., XIAP and GADD45B) and downregulation of proapoptotic genes (e.g., CASP8 and APAF1) in infected PMN. Transcript and protein levels of inflammation- and immunity-related genes were also altered. Upon interaction with PMN, G. bethesdensis altered the expression of ROS resistance genes in the presence of normal but not CGD PMN. Levels of bacterial stress response genes, including the ClpB gene, increased during phagocytosis by both normal and CGD PMN demonstrating responses to oxygen-independent PMN antimicrobial systems. Antisense knockdown demonstrated that ClpB is dispensable for extracellular growth but is essential for bacterial resistance to both normal and CGD PMN. Metabolic adaptation of Granulibacter growth in PMN included the upregulation of pyruvate dehydrogenase. Pharmacological inhibition of pyruvate dehydrogenase by triphenylbismuthdichloride was lethal to Granulibacter. This study expands knowledge of microbial pathogenesis of Granulibacter in cells from permissive (CGD) and nonpermissive (normal) hosts and identifies potentially druggable microbial factors, such as pyruvate dehydrogenase and ClpB, to help combat this antibiotic-resistant pathogen.
Laboratory of Clinical Infectious Diseases, NIAID, NIH, Bethesda, Maryland, USA Departments of Medicine and Microbiology, University of Texas Southwestern Medical Center, Dallas, Texas, USA.
David E
DE
Greenberg
Genomics Unit, Research Technologies Branch, Rocky Mountain Laboratories, NIAID, NIH, Hamilton, Montana, USA.
Daniel E
DE
Sturdevant
Laboratory of Clinical Infectious Diseases, NIAID, NIH, Bethesda, Maryland, USA Departments of Medicine and Microbiology, University of Texas Southwestern Medical Center, Dallas, Texas, USA.
Kimberly R
KR
Marshall-Batty
Laboratory of Host Defenses, NIAID, NIH, Bethesda, Maryland, USA.
Jessica
J
Chu
Laboratory of Host Defenses, NIAID, NIH, Bethesda, Maryland, USA.
Anthony M
AM
Pettinato
Genomics Unit, Research Technologies Branch, Rocky Mountain Laboratories, NIAID, NIH, Hamilton, Montana, USA.
Kimmo
K
Virtaneva
Genomics Unit, Research Technologies Branch, Rocky Mountain Laboratories, NIAID, NIH, Hamilton, Montana, USA.
John
J
Lane
Department of Microbiology, Oregon State University, Corvallis, Oregon, USA.
Bruce L
BL
Geller
Genomics Unit, Research Technologies Branch, Rocky Mountain Laboratories, NIAID, NIH, Hamilton, Montana, USA.
Stephen F
SF
Porcella
Laboratory of Host Defenses, NIAID, NIH, Bethesda, Maryland, USA.
John I
JI
Gallin
Laboratory of Clinical Infectious Diseases, NIAID, NIH, Bethesda, Maryland, USA.
Steven M
SM
Holland
Laboratory of Host Defenses, NIAID, NIH, Bethesda, Maryland, USA kzarember@niaid.nih.gov.
Kol A
KA
Zarember
D016947:Acetobacteraceae / Q000235:genetics* / Q000378:metabolism; D000328:Adult; D000368:Aged; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism; D005260:Female; D020869:Gene Expression Profiling; D006105:Granulomatous Disease, Chronic / Q000235:genetics* / Q000276:immunology / Q000382:microbiology; D064368:Healthy Volunteers; D054884:Host-Pathogen Interactions; D006801:Humans; D008297:Male; D008875:Middle Aged; D009504:Neutrophils / Q000378:metabolism* / Q000382:microbiology; D010587:Phagocytosis; D055815:Young Adult
D001426:Bacterial Proteins