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H-2D antigens released by thymocytes and cell adhesion

A S Curtis, M D Davies

    Journal of Immunogenetics
    |October 1, 1981
    PubMed
    Summary

    Researchers purified Thymocyte Interaction Modulation Factor (TIMF), identifying it as a soluble H-2D antigen fragment. This finding advances understanding of immune cell interactions and antigen presentation.

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    Area of Science:

    • Immunology
    • Molecular Biology
    • Biochemistry

    Background:

    • Previous purification methods for Thymocyte Interaction Modulation Factor (TIMF) resulted in preparations with multiple protein bands.
    • The precise identity and molecular characteristics of TIMF were not fully elucidated.

    Purpose of the Study:

    • To achieve the complete purification and definitive identification of mouse Thymocyte Interaction Modulation Factor (TIMF).
    • To characterize TIMF's biochemical properties and its relationship to known H-2 antigens.

    Main Methods:

    • Utilized silver-stained Polyacrylamide Gel Electrophoresis (PAGE) for protein analysis.
    • Employed monoclonal antibody affinity chromatography using anti-H-2Dk (15.5.5) and anti-H-2Kk (11.4.1) columns.
    • Determined molecular weight via PAGE analysis.
    • Assessed TIMF activity using an improved assay and investigated its inactivation by enzymes and inhibitors.

    Main Results:

    • Purified TIMF resolved as a single band on PAGE at approximately 34,000 Daltons.
    • TIMF activity specifically bound to anti-H-2Dk antibody columns, but not anti-H-2Kk columns.
    • Established evidence linking TIMF activity to the H-2D locus.
    • Demonstrated TIMF inactivation by specific enzymes and serine-esterase inhibitors.
    • Observed TIMF's effects on syngeneic leukocytes and macrophages without significant toxicity.

    Conclusions:

    • Thymocyte Interaction Modulation Factor (TIMF) is identified as a soluble fragment of the H-2D antigen.
    • TIMF is biochemically equivalent to a papainized H-2 fragment.
    • The study provides a refined method for TIMF activity assay and characterization.

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