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  • Title: Monocyte cytotoxicity: evidence for multiple mechanisms of in vitro erythrocyte target killing. Trypan blue can both inhibit and enhance target lysis.
    Author: Hall RE, Muchmore AV, Blaese RM.
    Journal: J Reticuloendothel Soc; 1982 Sep; 32(3):233-46. PubMed ID: 7153988.
    Abstract:
    Four systems of human monocyte-mediated cytotoxicity against chicken erythrocyte targets were studied using a panel of cellular inhibitors. Monocyte synergistic cytotoxicity (MSC), spontaneous monocyte-mediated cytotoxicity (SMMC), antibody-dependent cellular cytotoxicity (ADCC), and mitogen-induced cellular cytotoxicity (MICC) each had a different pattern of inhibition with these agents. Monocyte synergistic cytotoxicity, a system in which monocytes and certain complement components cooperate in the lysis of target cells, was inhibited only by trypan blue, a known inhibitor of lysosomal enzymes and macrophage cytolysis of tumor cells. In marked contrast, spontaneous monocyte-mediated cytotoxicity, a system in which monocytes become spontaneously cytotoxic after a 5- to 10-day incubation in culture, was enhanced by trypan blue, whereas MICC and ADCC were not affected. Agents which deplete cellular hydrogen peroxide and superoxide were most inhibitory for SMMC and had no effect on ADCC, MICC, and MSC. Inhibitors of protein synthesis only blocked MICC, having no effect on the other systems. These data support the hypothesis that monocytes possess multiple mechanisms by which they may inflict damage on erythroid targets, which may depend on oxidative intermediates (SMMC), protein synthesis (MICC), microfilament function (SMMC, MICC, ADCC), in addition to other mechanisms which have not yet been clearly defined (MSC). Trypan blue can thus both inhibit and enhance different forms of monocyte-mediated cytotoxicity. Further studies with this agent revealed that preincubation of cells with trypan blue depleted the ability of the cells to kill by SMMC but not by ADCC or MICC. Such studies suggest that trypan blue, like certain other compounds, has more than one mechanism of action on cellular processes important for the mediation of cytotoxicity.
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