Isolation and characterization of a T lymphocyte-derived differentiation inducing factor for the myeloid leukemic cell line HL-60.
Mitogen-stimulated mononuclear blood cells produce differentiation inducing factors (DIFs) for the promyelocytic cell line HL-60. We report that DIF is produced constitutively by a malignant T lymphocyte line HUT-102. DIF was purified 7,000-fold from HUT-102 conditioned media by utilizing ion-exchange chromatography with DEAE-Sepharose, gel chromatography, Blue-Sepharose chromatography, and preparative SDS-polyacrylamide gel electrophoresis (SDS-PAGE). The final preparation is susceptible to protease treatment, has a molecular weight of 46,000, as determined by SDS-PAGE and approximately 55,000 by gel filtration, has an isoelectric point of approximately 5.2, does not adhere to lectin-Sepharose and is resistant to periodate oxidation, and is free of colony-stimulating factor. DIF induced maturation of HL-60 into phagocytizing nitro blue tetrazolium reducing cells with the morphological characteristics of myelomonocytic or monocyte-like cells. An activity, co-chromatographing with DIF, acts synergistically with retinoic acid to induce maturation not only of HL-60, but also of the monoblast-like cell line U-937 (measured as percentage of cells reducing NBT).
D002460:Cell Line; D002471:Cell Transformation, Neoplastic / Q000187:drug effects / Q000378:metabolism / Q000473:pathology*; D002847:Chromatography, Agarose; D004355:Drug Stability; D004357:Drug Synergism; D006801:Humans; D015470:Leukemia, Myeloid, Acute / Q000378:metabolism* / Q000473:pathology; D008222:Lymphokines / Q000032:analysis / Q000302:isolation & purification* / Q000502:physiology; D009580:Nitroblue Tetrazolium; D013601:T-Lymphocytes / Q000378:metabolism*; D014212:Tretinoin / Q000494:pharmacology
D008222:Lymphokines; C050979:monocyte-macrophage differentiation factor; D009580:Nitroblue Tetrazolium; D014212:Tretinoin