Evidence for a factor in normal human serum that induces human neutrophilic granulocyte end-stage maturation in vitro.
The end-stage maturation of neutrophilic granulocyte precursor cells isolated from normal human bone marrow by Ficoll density centrifugation was studied in a liquid culture assay system used previously to study the maturation of guinea pig granulocyte precursors. Dialyzed normal human serum induced end-stage morphological maturation of human granulocyte precursors and this induction was proportional to a serum level of up to 5.0% in the assay medium. At serum concentrations greater than 5.0% a pronounced inhibition of maturation was observed. Passage of serum through a DEAE-Fractogel 650S column equilibrated with 0.01 M phosphate buffer (pH 7.0) resulted in the binding of the end-stage granulocyte maturation factor to the column. The activity eluted from the column in a fraction containing 17% of the starting serum protein that was inhibitor-free and was also capable of inducing the appearance of granulocyte alkaline phosphatase, a specific biochemical marker for granulocyte end-stage maturation. GMF is most likely a protein since it was destroyed by protease digestion. The data also indicate that neither purified human transferrin nor human recombinant granulocyte colony-stimulating factor can substitute for human serum GMF as a granulocyte end-stage maturation factor in this assay system. It was observed, however, that purified human transferrin greatly potentiated the effect of GMF suggesting that transferrin plays a supporting role in the end-stage maturation of human granulocytes in vitro. To our knowledge the evidence presented here indicates for the first time the existence of a neutrophilic granulocyte end-stage maturation factor in normal human serum.
Evans
W H
WH
Laboratory of Biochemistry, National Cancer Institute, Bethesda, MD.
Wilson
S M
SM
Bednarek
J M
JM
Peterson
E A
EA
Knight
R D
RD
Mage
M G
MG
McHugh
L
L
D000469:Alkaline Phosphatase / Q000032:analysis; D001685:Biological Factors / Q000097:blood* / Q000302:isolation & purification / Q000494:pharmacology; D015415:Biomarkers / Q000032:analysis; D001854:Bone Marrow Cells; D002469:Cell Separation; D002478:Cells, Cultured; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D003115:Colony-Stimulating Factors / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D016179:Granulocyte Colony-Stimulating Factor; D006098:Granulocytes / Q000166:cytology* / Q000187:drug effects; D006412:Hematopoietic Stem Cells / Q000166:cytology* / Q000187:drug effects; D006801:Humans; D009504:Neutrophils / Q000166:cytology* / Q000187:drug effects; D011994:Recombinant Proteins / Q000494:pharmacology; D012016:Reference Values; D014168:Transferrin / Q000494:pharmacology
D001685:Biological Factors; D015415:Biomarkers; D003115:Colony-Stimulating Factors; D011994:Recombinant Proteins; D014168:Transferrin; D016179:Granulocyte Colony-Stimulating Factor; D000469:Alkaline Phosphatase