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Concanavalin A-induced macrophage aggregating factor

P Badenoch-Jones

    The Australian Journal of Experimental Biology and Medical Science
    |October 1, 1981
    PubMed
    Summary
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    Researchers quantified macrophage aggregating factor (MAgF) production using spectrophotometry, not platelet aggregometry. This lymphokine activity was characterized by molecular weight and confirmed to be MAgF, not Concanavalin A.

    Area of Science:

    • Immunology
    • Cell Biology
    • Biochemistry

    Background:

    • Lymphokines are signaling molecules crucial for immune responses.
    • Macrophage aggregating factor (MAgF) plays a role in cellular aggregation.
    • Previous assays for MAgF relied on less precise methods like platelet aggregometry.

    Purpose of the Study:

    • To investigate the production of macrophage aggregating factor (MAgF) by Concanavalin A (Con A)-pulsed guinea-pig spleen cells.
    • To establish a quantitative assay for MAgF.
    • To characterize the properties of MAgF, including its molecular weight and specificity.

    Main Methods:

    • Quantitative assay of MAgF using spectrophotometric measurement of peritoneal exudate cell (PEC) light absorbance.
    • Gel filtration chromatography (Sephadex G-200) to determine molecular weight.

    Related Experiment Videos

  • Inhibitor studies using alpha-methyl-D-mannoside to differentiate MAgF activity from Con A.

    • Main Results:

    • A quantitative spectrophotometric assay for MAgF was developed, replacing the Born platelet aggregometer.
    • Both Con A- and antigen-induced MAgF were found to elute from Sephadex G-200 in the 30,000-70,000 m wt range.
    • Inhibitor studies confirmed that the observed PEC aggregation was mediated by MAgF, not residual Con A.

    Conclusions:

    • A robust and quantitative assay for MAgF has been established.
    • MAgF is a lymphokine with a molecular weight between 30,000-70,000 m wt.
    • The study successfully differentiated MAgF-induced cell aggregation from Con A-induced aggregation.