A human myeloid cell line producing stem cell growth factor, KPB-M15, secretes another growth factor active on murine hematopoietic progenitor cells.
Human stem cell growth factor (SCGF) produced by a myeloid cell line, KPB-M15, exhibits species-specific hematopoietic activities. However, KPB-M15-conditioned medium induced colony formation of mouse bone marrow cells. KPB-M15-derived colony-stimulating activity (CSA) was purified through Butyl-Toyopearl 650c and Cu2+ chelating-Sepharose 6B chromatography. TSK-G3000SW gel filtration of the purified preparation presented 3 distinct peaks around Vo, 150 kD and 85 kD. Gel fractions extracted from SDS-PAGE had macrophage colony-stimulating factor (M-CSF)-specific amino acid sequences. PCR, Northern hybridization and ELISA demonstrated that KPB-M15 cells secreted a significant amount of M-CSF and IL-6. Anti-M-CSF but not anti-IL-6 antibody abrogated CSA in KPB-M15-CM. IL-6 hardly synergized with M-CSF to enhance colony formation. Collectively, M-CSF is a sole CSA for murine hematopoietic progenitor cells in KPB-M15-CM. This is the first report of a human myeloid cell line, KPB-M15, constitutively producing M-CSF in addition to SCGF and IL-6. It can be useful in investigating the mechanism of production of M-CSF.
Hiraoka
A
A
Department of Internal Medicine, Osaka Dental University, Osaka, Japan. hestroboy@miyavision.ne.jp
Nagasawa
T
T
Ohta
N
N
Sugimura
A
A
Shimizu
N
N
Ooi
K
K
Shimizu
S
S
D000818:Animals; D000906:Antibodies / Q000494:pharmacology; D001854:Bone Marrow Cells / Q000187:drug effects / Q000378:metabolism*; D003115:Colony-Stimulating Factors / Q000096:biosynthesis* / Q000302:isolation & purification / Q000494:pharmacology*; D017077:Culture Media, Conditioned / Q000378:metabolism / Q000494:pharmacology; D016207:Cytokines / Q000096:biosynthesis; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D006412:Hematopoietic Stem Cells / Q000166:cytology* / Q000187:drug effects; D006801:Humans; D015850:Interleukin-6 / Q000276:immunology / Q000494:pharmacology; D016173:Macrophage Colony-Stimulating Factor / Q000276:immunology / Q000494:pharmacology; D051379:Mice; D008810:Mice, Inbred C57BL; D012967:Sodium Dodecyl Sulfate; D019089:Stem Cell Factor / Q000096:biosynthesis*; D014407:Tumor Cells, Cultured