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Proliferation and Differentiation of Murine Myeloid Precursor 32D/G-CSF-R Cells
Published on: February 21, 2018
Two separate differentiation inducing proteins for human myeloid leukemia cells and their isolation from normal
M Abolhassani1, W M Riley, E Feldman
1Department of Medicine, New York Medical College, Valhalla 10595.
Insights
Normal lymphocytes contain a maturation inducer that differentiates leukemia cells. This inducer, a 54 kDa protein, is less abundant in acute myelogenous leukemia patients, suggesting a role in leukemia cell differentiation.
Area of Science:
- Immunology
- Cell Biology
- Hematology
Background:
- Peripheral blood lymphocytes (PBL) conditioned medium contains activity promoting myeloid leukemia cell differentiation.
- This activity induces differentiation of human myeloid leukemia cells into monocytes and macrophages.
Purpose of the Study:
- To isolate and characterize the maturation inducer activity from stimulated normal PBL.
- To investigate the role of this inducer in acute myelogenous leukemia (AML).
Main Methods:
- Isolation of maturation inducer activity using salt precipitation, chromatography (Sepharose CL-6B ion exchange and affinity), and electrophoresis.
- Characterization of molecular weight (MW) and identification using ELISA and differentiation blocking assays.
- Detection of the 54 kDa protein on PBL surface membranes via immunofluorescent antibody technique.
Main Results:
- Two distinct MW activities (52-56 kDa and 32-35 kDa) were identified, mediating HL-60 promyelocyte differentiation.
- The higher MW species was identified as a 54 kDa polypeptide, distinct from IL-3 and IL-6.
- The 32-35 kDa activity was neutralized by anti-IL-3 antibodies.
- Significantly lower proportions and numbers of maturation inducer-bearing lymphocytes were found in AML patients compared to healthy donors.
Conclusions:
- A 54 kDa protein on PBL surface membranes acts as a maturation inducer for myeloid leukemia cells.
- Reduced levels of this inducer in AML patients suggest its potential role in impaired leukemia cell differentiation.
- Further research into these physiological factors could offer insights into leukemia treatment strategies.
Abstract:
Conditioned medium from mitogen stimulated normal peripheral blood lymphocytes (PBL) has been demonstrated to contain a maturation inducer activity mediating the differentiation of human myeloid leukemia cells to monocytes and macrophages. The maturation inducer activity was isolated by salt precipitation, Sepharose CL-6B ion exchange and affinity chromatographies and electrophoresis. Two separate activities with M.W. ranges of 52-56 and 32-35 kDa capable of mediating the terminal differentiation of leukemic HL-60 promyelocytes to monocytes and macrophages were detected. The higher molecular weight species was determined to be a 54 kDa single polypeptide and was found to be distinct from IL-3 and IL-6 by ELISA and differentiation blocking assay. The inducing activity of the 32-35 kDa material was largely neutralized after treatment with anti-IL-3, but not with other antibodies. Employing the immunofluorescent antibody technique, the 54 kDa protein was detected on the surface membranes of PBL. The proportions and number of maturation inducer bearing lymphocytes in patients with acute myelogenous leukemia (0.4% and 35/mm3, respectively) were significantly lower than that of healthy donors (7.9% and 178/mm3) The role of these physiological factors in leukemia cell differentiation is discussed.
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