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Related Experiment Videos

K562 killing by K, IL 2-responsive NK, and T cells involves different effector cell post-binding trigger mechanisms.

F Shanahan, M D Brogan, W Newman

    Journal of Immunology (Baltimore, Md. : 1950)
    |July 15, 1986
    PubMed
    Summary

    Monoclonal antibody 13.3 reveals distinct trigger mechanisms in Natural Killer (NK) cell-mediated K562 lysis. This antibody highlights the heterogeneity in cytotoxic pathways, even after interleukin-2 activation of lymphocytes.

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    Area of Science:

    • Immunology
    • Cellular Biology
    • Cytotoxicity Mechanisms

    Background:

    • Natural Killer (NK) cells and other lymphocyte populations mediate cytotoxicity against K562 target cells.
    • The precise trigger mechanisms involved in NK-K562 lysis and other cytotoxic pathways remain incompletely understood.
    • Monoclonal antibody 13.3 has been identified as a specific blocker of a post-binding effector cell trigger in the NK-K562 cytolytic sequence.

    Purpose of the Study:

    • To utilize monoclonal antibody 13.3 to delineate differences in the lytic trigger processes of NK cells and other mechanisms of K562 lysis.
    • To investigate the effect of antibody 13.3 on various lymphocyte-mediated cytotoxic activities against K562 cells.
    • To examine the impact of interleukin-2 (IL-2) activation on the sensitivity of cytotoxic pathways to antibody 13.3.

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    Main Methods:

    • Employing monoclonal antibody 13.3 to block specific cytotoxic pathways.
    • Assessing the inhibition of K562 cell lysis by freshly isolated peripheral blood lymphocytes (PBL) and purified large granular lymphocytes (LGL).
    • Evaluating the effect of antibody 13.3 on antibody-dependent cell-mediated cytotoxicity (ADCC) and lectin-dependent cellular cytotoxicity (LDCC).
    • Culturing PBL with interleukin-2 (IL-2) and subsequently testing their sensitivity to antibody 13.3.

    Main Results:

    • Monoclonal antibody 13.3 specifically inhibited NK-mediated lysis of K562 cells at a post-binding effector cell level.
    • Antibody 13.3 did not inhibit antibody-dependent cell-mediated cytotoxicity (ADCC) or lectin-dependent cellular cytotoxicity (LDCC) against K562 cells.
    • The inhibitory effect of antibody 13.3 on NK-K562 lysis persisted in IL-2 activated PBL for up to 48 hours.
    • A diminishing inhibition was observed in fetal calf serum-containing media after 48 hours, but not in autologous serum.
    • 13.3-unresponsive lytic activity in cultured PBL was found in both LGL and T cell fractions, indicating heterogeneity.

    Conclusions:

    • K562 lysis by freshly isolated human lymphocytes involves heterogeneous post-binding effector cell trigger mechanisms for NK, K, and LDCC.
    • Lymphocyte-mediated K562 lysis following IL-2 culture also exhibits heterogeneity in its trigger mechanisms.
    • Monoclonal antibody 13.3 serves as a valuable tool for dissecting distinct cytotoxic pathways and their regulation.