Monocyte-derived recruiting activity: Kinetics of production and effects of endotoxin

E. McCall, G. C. Bagby

    Research output: Contribution to journalArticle

    6 Citations (Scopus)

    Abstract

    Cultured monocytes release a factor, monocyte-derived recruiting activity (MRA), which stimulates fibroblasts, endothelial cells, and T lymphocytes to produce colony-stimulating activity (CSA). We studied the kinetics of MRA production using a technique in which MRA levels were measured in a two stage bioassay. We used umbilical vein endothelial cells as the MRA-responsive granulocyte-macrophage (CFU-GM)-enriched bone marrow cells (T lymphocyte- and monocyte-depleted, low density bone marrow cells) as the CSA-responsive cells. MRA stimulated a 30-fold increase in CSA production by endothelial cells. MRA production was detected in supernatants from as few as 103 monocytes per milliliter, required the presence of fetal calf serum, and was inhibited by cycloheximide (10 to 100 μg/mL) and puromycin (10 to 50 μg/mL). Production was detectable after 24 hours of monocyte incubation, was maintained for 3 days, and fell to undetectable levels by 7 days. With the addition of bacterial endotoxin (lipopolysaccharide [LPS]) (50 μg per 106 cells), MRA was detectable after only 3 hours of incubation, and levels peaked at 24 hours. Further, maximum MRA levels in the supernatants of LPS-stimulated monocytes were up to 10 times greater than peak levels in the supernatants of unstimulated monocytes. Endotoxin augmented monocyte production of MRA to a greater extent than it did CSA production, indicating that the stimulation of CSA production by endotoxin may be at least partly indirect. The responsiveness of MRA production to endotoxin in vitro is consistent with the notion that MRA may be a biologically relevant regulator of CSA production by cells of the hematopoietic microenvironment.

    Original languageEnglish (US)
    Pages (from-to)689-695
    Number of pages7
    JournalBlood
    Volume65
    Issue number3
    StatePublished - 1985

    Fingerprint

    Endotoxins
    Monocytes
    Kinetics
    Endothelial cells
    T-cells
    Lipopolysaccharides
    Bone
    Cells
    Puromycin
    Endothelial Cells
    Bioassay
    Macrophages
    Fibroblasts
    Cycloheximide
    Bone Marrow Cells
    T-Lymphocytes
    Cellular Microenvironment
    Granulocyte-Macrophage Progenitor Cells
    Umbilical Veins
    Granulocytes

    ASJC Scopus subject areas

    • Hematology

    Cite this

    Monocyte-derived recruiting activity : Kinetics of production and effects of endotoxin. / McCall, E.; Bagby, G. C.

    In: Blood, Vol. 65, No. 3, 1985, p. 689-695.

    Research output: Contribution to journalArticle

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    abstract = "Cultured monocytes release a factor, monocyte-derived recruiting activity (MRA), which stimulates fibroblasts, endothelial cells, and T lymphocytes to produce colony-stimulating activity (CSA). We studied the kinetics of MRA production using a technique in which MRA levels were measured in a two stage bioassay. We used umbilical vein endothelial cells as the MRA-responsive granulocyte-macrophage (CFU-GM)-enriched bone marrow cells (T lymphocyte- and monocyte-depleted, low density bone marrow cells) as the CSA-responsive cells. MRA stimulated a 30-fold increase in CSA production by endothelial cells. MRA production was detected in supernatants from as few as 103 monocytes per milliliter, required the presence of fetal calf serum, and was inhibited by cycloheximide (10 to 100 μg/mL) and puromycin (10 to 50 μg/mL). Production was detectable after 24 hours of monocyte incubation, was maintained for 3 days, and fell to undetectable levels by 7 days. With the addition of bacterial endotoxin (lipopolysaccharide [LPS]) (50 μg per 106 cells), MRA was detectable after only 3 hours of incubation, and levels peaked at 24 hours. Further, maximum MRA levels in the supernatants of LPS-stimulated monocytes were up to 10 times greater than peak levels in the supernatants of unstimulated monocytes. Endotoxin augmented monocyte production of MRA to a greater extent than it did CSA production, indicating that the stimulation of CSA production by endotoxin may be at least partly indirect. The responsiveness of MRA production to endotoxin in vitro is consistent with the notion that MRA may be a biologically relevant regulator of CSA production by cells of the hematopoietic microenvironment.",
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