TY - JOUR
T1 - Human colony-stimulating factor (CSF-1) radioimmunoassay
T2 - Resolution of three subclasses of human colony-stimulating factors
AU - Das, S. K.
AU - Stanley, E. R.
AU - Guilbert, L. J.
AU - Forman, L. W.
PY - 1981
Y1 - 1981
N2 - The partial purification of human urinary CSF-1 to approximately 4 x 106 U/mg protein from large volumes of human urine is described. Carrier-free iodination of the partially purified preparation and subsequent receptor binding and elution of the resulting 125I-CSF-1 yielded 125I-CSF-1, which, by several criteria, was free of contaminating 125I-proteins. The 125I-CSF-1 (approximately 4 x 107 U/mg of iodinated protein) was biologically active and suitable for use in target cell binding studies. It was used to develop a RIA that specifically detects the human CSF subclass that stimulates macrophage production (CSF-1). This RIA, the CSF-1 radioreceptor assay, and the murine and human CSF bioassays were used in comparative studies of a variety of CSF preparations. Results of these studies indicate that there are at least three human CSF subclasses: CSF-1 which stimulates the formation of colonies containing macrophages by murine and human hemopoietic cells; another subclass stimulating the formation of colonies (possibly containing neutrophils, eosinophils, and macrophages) by human but not murine hemopoietic cells; and a third subclass that stimulates the formation of colonies (possibly containing neutrophils and macrophages) by both human and murine hemopoietic cells. Anti-human urinary CSF-1 antibody selectively prevented the development of human bone marrow colonies with the gross morphology of those stimulated by purified human urinary CSF-1, in cultures stimulated by a variety of CSF preparations. The average level of CSF-1 in the serum of 14 normal adult humans, as determined by RIA, was 174 ± 76 U/ml (approximately 40 pM).
AB - The partial purification of human urinary CSF-1 to approximately 4 x 106 U/mg protein from large volumes of human urine is described. Carrier-free iodination of the partially purified preparation and subsequent receptor binding and elution of the resulting 125I-CSF-1 yielded 125I-CSF-1, which, by several criteria, was free of contaminating 125I-proteins. The 125I-CSF-1 (approximately 4 x 107 U/mg of iodinated protein) was biologically active and suitable for use in target cell binding studies. It was used to develop a RIA that specifically detects the human CSF subclass that stimulates macrophage production (CSF-1). This RIA, the CSF-1 radioreceptor assay, and the murine and human CSF bioassays were used in comparative studies of a variety of CSF preparations. Results of these studies indicate that there are at least three human CSF subclasses: CSF-1 which stimulates the formation of colonies containing macrophages by murine and human hemopoietic cells; another subclass stimulating the formation of colonies (possibly containing neutrophils, eosinophils, and macrophages) by human but not murine hemopoietic cells; and a third subclass that stimulates the formation of colonies (possibly containing neutrophils and macrophages) by both human and murine hemopoietic cells. Anti-human urinary CSF-1 antibody selectively prevented the development of human bone marrow colonies with the gross morphology of those stimulated by purified human urinary CSF-1, in cultures stimulated by a variety of CSF preparations. The average level of CSF-1 in the serum of 14 normal adult humans, as determined by RIA, was 174 ± 76 U/ml (approximately 40 pM).
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U2 - 10.1182/blood.v58.3.630.630
DO - 10.1182/blood.v58.3.630.630
M3 - Article
C2 - 6973347
AN - SCOPUS:0019373405
SN - 0006-4971
VL - 58
SP - 630
EP - 641
JO - Blood
JF - Blood
IS - 3
ER -