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The induction of specific macrophage cytotoxicity by allo-sensitized T cells in vitro is shown to be independent of the presence of lipopolysaccharide (LPS). This contrasts with the induction of macrophage cytotoxicity by Macrophage Activating Factor (MAF). The specific macrophage cytotoxicity could be induced in LPS-free medium (less than 1 ng/ml). Addition of LPS to the macrophages did not increase the cytotoxicity. Addition of LPS-binding polymyxin B to the macrophages before and during the induction of cytotoxicity did not reduce the specific macrophage cytotoxicity. Macrophages obtained from the LPS-unresponsive mouse strain C3H/Hej were rendered cytotoxic by the allo-sensitized lymphocytes to the same extent as the macrophages from LPSresponsive C3HeB/Fe and C57BL mice. This indicates that the induction of macrophage cytotoxicity by MAF is different from the induction of specific macrophage cytotoxicity by Specific Macrophage Arming Factor (SMAF).

Specific T cell factors, such as specific macrophage arming factor (SMAF), are involved in the initiation of the immune response. Induction of SMAF-producing T lymphocytes in vivo and of SMAF production by T lymphocytes in vitro is dependent on the presence of intact tumor cells, and is independent of antigen presentation by macrophages. SMAF renders peritoneal macrophages cytotoxic for tumor cells. The armed peritoneal macrophages expressed a specific cytotoxicity. However, antigen-presenting cells can trigger lymphokine-producing T lymphocytes. These T lymphocytes produce lymphokines (e.g. macrophage activating factor (MAF] that activate macrophages. The MAF-activated macrophages express a non-specific tumoricidal activity. In the present study, we investigated the difference in the induction of macrophage cytotoxicity by SMAF and MAF. The following differences were found: 1) SMAF renders peritoneal resident macrophages cytotoxic, whereas MAF could only render peritoneal exudate macrophages cytotoxic. 2) SMAF requires only a 4-h incubation with macrophages, whereas MAF activates macrophages optimally after 12 h. 3) SMAF-armed macrophages recognize only the specific target cell(s), and thus, the cytotoxicity is specific in its expression. MAF activated macrophages were non-specifically cytotoxic. 4) Lipopolysaccharide (LPS) in the culture medium did not enhance the cytotoxicity of SMAF-armed macrophages. In contrast, MAF induced tumoricidal activity was enhanced by adding LPS to the culture medium. 5) After adsorption chromatography with anti-murine interferon-gamma (IFN-gamma), the arming capacity of SMAF supernatant was not reduced, whereas the activating capacity of the MAF supernatant was significantly reduced or abrogated. After immunization of mice with allogeneic tumor cells, SMAF-producing lymphocytes were detected in the draining lymph nodes already 4 days after immunization and up to 12 days. Lymphocytes with the capacity to produce MAF were present in the draining lymph nodes 14-24 days after immunization. Our data indicate that the T cell factors SMAF and MAF can both render macrophages cytotoxic, but act in a different way and during different stages of the cellular immune response against allogeneic tumor cells.

Spleen cells sensitized to tumour cells have an anti-tumour effect on injected syngeneic lymphosarcoma cells in mice. This study shows that this anti-tumour effect can be enhanced by induced peritoneal macrophages and by macrophage-like tumour cells (macrophages). Addition of macrophages to the intraperitoneally injected sensitized spleen cells stimulated the anti-tumour effect. This was observed both with intraperitoneally injected tumour cells and with subcutaneously transplanted tumour cells. The anti-tumour effect is the result of a cooperation between T cells and macrophages. In vitro incubation of immune T-cells with macrophages or macrophage-like cells enhanced the in vivo anti-tumour activity of the sensitized T-lymphocytes. Neither the presence of antigen nor the proliferation of the immune T-cells were a prerequisite to enhance this anti-tumour effect. Our experiments suggest that a macrophage factor is responsible for the enhancement of the anti-tumour effect. Based on the results of this paper and other studies we propose the following sequence of events to explain the anti-tumour effect of injected sensitized T-lymphocytes and macrophages: injected macrophages enhance the anti-tumour effect of sensitized lymphocytes. These stimulated lymphocytes migrate to the tumour located elsewhere and recognize the tumour antigens. Subsequently, the lymphocytes render (host) macrophages in the tumour cytotoxic to tumour cells.