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Related Experiment Videos

Lymphocyte binding of fluid phase mouse C3b.

M Papamichail, M B Pepys

    Immunology
    |March 1, 1979
    PubMed
    Summary

    Mouse complement protein C3b binds to lymphocytes. Aggregation of C3b is crucial for detection on mouse cells, revealing new insights into complement receptor interactions and immunohistochemistry.

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    Area of Science:

    • Immunology
    • Complement System Biology

    Background:

    • Mouse complement protein C3b can bind to both human and mouse lymphocytes.
    • Standard immunofluorescence techniques struggle to detect C3b bound to mouse cells.

    Purpose of the Study:

    • To investigate the binding characteristics of mouse C3b to lymphocytes.
    • To optimize methods for detecting C3b-lymphocyte interactions.
    • To understand the implications for immunohistochemical studies.

    Main Methods:

    • Utilized fluorescein-conjugated mouse C3b (FITC-mouse C3b) for direct staining.
    • Investigated the effect of C3b aggregation on binding detection.
    • Assessed C3b-induced lymphocyte responses like patching, capping, and endocytosis.
    • Examined the inhibitory effects of various agents on these cellular responses.

    Main Results:

    • Mouse C3b binds to both human and mouse lymphocytes, but detection on mouse cells requires C3b aggregation.
    • Aggregated FITC-mouse C3b effectively stains lymphocytes and induces receptor-mediated endocytosis.
    • The failure to detect non-aggregated C3b on mouse cells is likely due to receptor inaccessibility to anti-C3 antibodies.
    • Cellular responses to aggregated C3b-FITC are sensitive to temperature, metabolic inhibitors, and cytoskeletal disruptors.

    Conclusions:

    • C3b aggregation is essential for its immunofluorescent detection on mouse lymphocytes.
    • The binding and cellular response mechanisms of C3b to lymphocyte complement receptors are elucidated.
    • Findings suggest limitations in current immunohistochemical methods for detecting in vivo C3 binding to lymphoid cells.

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