Tumor Growth Inhibitory and Natural Suppressor Activities of Murine Bone Marrow Cells: A Comparative Study

Abstract

The murine bone marrow (BM) cells having a certain phenotypic similarity to null natural suppressor (NS) cells have been previously established to be able to inhibitin vitroleukemic cell growth in a genetically unrestricted manner. In this study we found that the treatment of normal (C57BL/6 × DBA)F1 BM cells with a lysosomotropic agent,l-leucine methyl ester (LME), largely abrogated their ability to reduce both P815 mastocytoma and L1210 lymphoma cell proliferation, as well as their NS activity tested for suppression of mitogen (Con A or LPS)-driven spleen cell proliferation. However, after being depleted of the cells binding wheat germ agglutinin (WGA), the BM cells maintained tumor growth-inhibitory activity, while demonstrating no significant NS activity. Moreover, in contrast to T-cell blastogenesis-inhibitory NS activity of BM cells, that was greatly reduced by the addition into the culture of either neutralizing anti-interferon (IFN)-γ antibody (Ab) orNG-monomethyl-l-arginine, a competitive inhibitor of NO synthase, natural antitumor cytostatic activity of BM cells was not found to be dependent on the presence in medium of IFN-γ and to be associated with NO production. When incubated at suboptimal numbers with tumor cells on conic, round, and flat well bottoms for 7 h, BM cells provided the most, middle, and least (or no) tumor growth inhibition, respectively, suggesting, thereby, a significance of cell density in cytostatic process. It was also found that the BM cells cultured for 20 h with the medium conditioned by mitogen-preactivated T or B lymphocytes were significantly more suppressive to tumor cell proliferation than the BM cells cultured in medium alone. The potentiation of BM-cell cytostatic activity by T-cell-conditioned medium (CM), but not that by B-cell-CM, was found to be partially reversed by anti-IFN-γ Ab. Finally, a noticeable tumor growth-inhibitory activity, which could be significantly enhanced upon T-cell-CM, was shown to be also attributable to BM cells from athymic BALB/c mice. Taken together, the results suggest that (1) the tumor growth inhibitory BM cells and the NS BM cells are not identical in their cell compositions, but also differ in their mechanisms of antiproliferative action; (2) a contact cell-to-cell interaction may play a significant role in BM-cell-mediated tumor growth inhibition; (3) the activated lymphocytes, through both IFN-γ-mediated and IFN-γ-independent pathways, are able to operatively up-regulate the cytostatic activity of BM cells; and (4) the tumor growth-inhibitory activity exhibited by the normal unmanipulated BM cells, at least in its significant part, may not be a consequence of thymus-dependent immune processes occurring normally in the body.