Neutropenia with impaired immune response to Streptococcus pneumoniae in ceramide kinase-deficient mice.
In mammals, ceramide kinase (CerK)-mediated phosphorylation of ceramide is the only known pathway to ceramide-1-phosphate (C1P), a recently identified signaling sphingolipid metabolite. To help delineate the roles of CerK and C1P, we knocked out the gene of CerK in BALB/c mice by homologous recombination. All in vitro as well as cell-based assays indicated that CerK activity is completely abolished in Cerk-/- mice. Labeling with radioactive orthophosphate showed a profound reduction in the levels of de novo C1P formed in Cerk-/- macrophages. Consistently, mass spectrometry analysis revealed a major contribution of CerK to the formation of C16-C1P. However, the significant residual C1P levels in Cerk-/- animals indicate that alternative routes to C1P exist. Furthermore, serum levels of proapoptotic ceramide in these animals were significantly increased while levels of dihydroceramide as the biosynthetic precursor were reduced. Previous literature pointed to a role of CerK or C1P in innate immune cell function. Using a variety of mechanistic and disease models, as well as primary cells, we found that macrophage- and mast cell-dependent readouts are barely affected in the absence of CerK. However, the number of neutrophils was strikingly reduced in blood and spleen of Cerk-/- animals. When tested in a model of fulminant pneumonia, Cerk-/- animals developed a more severe disease, lending support to a defect in neutrophil homeostasis following CerK ablation. These results identify ceramide kinase as a key regulator of C1P, dihydroceramide and ceramide levels, with important implications for neutrophil homeostasis and innate immunity regulation.
Graf
Christine
C
Novartis Institutes for BioMedical Research, Vienna, Austria.
Zemann
Barbara
B
Rovina
Philipp
P
Urtz
Nicole
N
Schanzer
Andrea
A
Reuschel
Roland
R
Mechtcheriakova
Diana
D
Müller
Matthias
M
Fischer
Evelin
E
Reichel
Claudia
C
Huber
Susanna
S
Dawson
Janet
J
Meingassner
Josef G
JG
Billich
Andreas
A
Niwa
Satoru
S
Badegruber
Rudolf
R
Van Veldhoven
Paul P
PP
Kinzel
Bernd
B
Baumruker
Thomas
T
Bornancin
Frédéric
F
D000818:Animals; D001169:Arthritis, Experimental / Q000201:enzymology / Q000276:immunology / Q000382:microbiology; D002478:Cells, Cultured; D002518:Ceramides / Q000378:metabolism; D005260:Female; D006967:Hypersensitivity / Q000201:enzymology / Q000235:genetics / Q000276:immunology; D007113:Immunity, Innate / Q000235:genetics; D017737:Macrophages, Peritoneal / Q000166:cytology / Q000201:enzymology / Q000276:immunology; D008297:Male; D008407:Mast Cells / Q000201:enzymology / Q000276:immunology / Q000382:microbiology; D051379:Mice; D008807:Mice, Inbred BALB C; D018345:Mice, Knockout; D009503:Neutropenia / Q000201:enzymology* / Q000235:genetics / Q000276:immunology* / Q000382:microbiology; D010323:Passive Cutaneous Anaphylaxis / Q000276:immunology; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000172:deficiency* / Q000235:genetics / Q000378:metabolism; D011018:Pneumonia, Pneumococcal / Q000201:enzymology* / Q000235:genetics / Q000276:immunology*; D013296:Streptococcus pneumoniae / Q000276:immunology*
D002518:Ceramides; C065576:ceramide 1-phosphate; C109343:dihydroceramide; D017853:Phosphotransferases (Alcohol Group Acceptor); C066324:ceramide kinase