ISOLATION AND CHARACTERIZATION OF A LYMPHOCYTE-T-DERIVED DIFFERENTIATION INDUCING FACTOR FOR THE MYELOID LEUKEMIC-CELL LINE HL-60

  • 1 January 1984
    • journal article
    • research article
    • Vol. 63  (3) , 510-517
Abstract
Mitogen-stimulated mononuclear blood cells produce differentiation inducing factors (DIF) for the promyelocytic cell line HL-60. DIF is produced constitutively by a malignant T lymphocyte line HUT-102. DIF was purified 7000-fold from HUT-102 conditioned media by utilizing ion-exchange chromatography with DEAE-Sepharose, gel chromatography, Blue-Sepharose chromatography, and preparative SDS[sodium-dodecyl sulfate]-polyacrylamide gel electrophoresis (SDS-PAGE). The final preparation is susceptible to protease treatment, has a MW 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 nitroblue tetrazolium [NBT] reducing cells with the morphological characteristics of myelomonocytic or monocyte-like cells. An activity, cochromatographing 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).

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