Human neutrophilic granulocyte end-stage maturation factor...

Details Human neutrophilic granulocyte end-stage maturation factor... Human neutrophilic granulocyte end-stage maturation factor...

C - Chemistry, Metallurgy

07

K

530/15.06

C07K 14/475 (2006.01) C07K 14/52 (2006.01) A61K 38/00 (2006.01) G01N 30/02 (2006.01)

Patent

CA 2003350

HUMAN NEUTROPHILIC GRANULOCYTE END-STAGE MATURATION FACTOR AND ITS PREPARATION AND USE ABSTRACT OF THE DISCLOSURE 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 0.01M phosphate buffer (pH 7.0) resulted in the binding of the end-stage granulocyte maturation factor (GMF) 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 phosphates, 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. Taken together the evidence presented here strongly supports the existence of a previously undescribed neutrophilic granulocyte end-stage maturation factor in normal human serum,

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