Neutrophils deficient in PU.1 do not terminally differentiate or become functionally competent.
PU.1 is an ets family transcription factor that is expressed specifically in hematopoietic lineages. Through gene disruption studies in mice we have previously shown that the expression of PU.1 is not essential for early myeloid lineage or neutrophil commitment, but is essential for monocyte/macrophage development. We have also shown that PU.1-null (deficient) neutrophils have neutrophil morphology and express neutrophil-specific markers such as Gr-1 and chloroacetate esterase both in vivo and in vitro. We now demonstrate that although PU.1-null mice develop neutrophils, these cells fail to terminally differentiate as shown by the absence of messages for neutrophil secondary granule components and the absence or deficiency of cellular responses to stimuli that normally invoke neutrophil function. Specifically, PU.1-deficient neutrophils fail to respond to selected chemokines, do not generate superoxide ions, and are ineffective at bacterial uptake and killing. The failure to produce superoxide could, in part, be explained by the absence of the gp91 subunit of nicotinamide adenine dinucleotide phosphate oxidase, as shown by our inability to detect messages for the gp91(phox) gene. Incomplete maturation of PU.1-deficient neutrophils is cell autonomous and persists in cultured PU.1-deficient cells. Our results indicate that PU.1 is not necessary for neutrophil lineage commitment but is essential for normal development, maturation, and function of neutrophils.
The Burnham Institute and the Department of Immunology, The Scripps Research Institute, La Jolla, CA, USA.
K L
KL
Anderson
K A
KA
Smith
F
F
Pio
B E
BE
Torbett
R A
RA
Maki
D000818:Animals; D002454:Cell Differentiation; D002478:Cells, Cultured; D002634:Chemotaxis, Leukocyte; D003594:Cytoplasmic Granules / Q000235:genetics; D004268:DNA-Binding Proteins / Q000378:metabolism* / Q000502:physiology; D016179:Granulocyte Colony-Stimulating Factor / Q000494:pharmacology; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000494:pharmacology; D050835:Interferon Regulatory Factors; D016209:Interleukin-8 / Q000494:pharmacology; D016173:Macrophage Colony-Stimulating Factor / Q000494:pharmacology; D051379:Mice; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000494:pharmacology; D019255:NADPH Oxidases / Q000235:genetics; D018375:Neutrophil Activation; D009504:Neutrophils / Q000166:cytology* / Q000502:physiology*; D010587:Phagocytosis; D010641:Phenotype; D012333:RNA, Messenger / Q000032:analysis; D013481:Superoxides / Q000378:metabolism; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D015534:Trans-Activators / Q000172:deficiency* / Q000378:metabolism* / Q000502:physiology; D000099218:Interferon Regulatory Factor-4
D004268:DNA-Binding Proteins; D016179:Granulocyte Colony-Stimulating Factor; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D050835:Interferon Regulatory Factors; D016209:Interleukin-8; D016173:Macrophage Colony-Stimulating Factor; D009240:N-Formylmethionine Leucyl-Phenylalanine; D019255:NADPH Oxidases; D012333:RNA, Messenger; D013481:Superoxides; D013755:Tetradecanoylphorbol Acetate; D015534:Trans-Activators; D000099218:Interferon Regulatory Factor-4