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Cloned human T-cell lines were produced by limiting dilution of lymphocytes alloactivated in mixed leukocyte cultures, followed by expansion of clonal progeny with interleukin 2. Selected clones were analyzed for cell-mediated cytotoxicity (CTX) against a variety of targets susceptible or resistant to lytic attack by natural killer (NK) cells. Clones with classical alloantigen-restricted lytic capacity for normal lymphoid targets were found to be distinct from those mediating natural killer-like CTX against NK-susceptible cell line targets. This was established by direct CTX assays and by cold target cross-competition experiments. Moreover, clones with NK-like activity displayed heterogeneous patterns of lysis on different target cell lines in direct CTX, suggesting a clonal distribution of receptors for NK-like target antigens amongst these cultured human NK-active T-cell clones.

Monoclonal antibodies were induced against leukemic T cells from a patient with chronic lymphocytic leukemia exhibiting natural killer (NK) activity. Two antibodies, termed T811 and M522, reacted by indirect immunofluorescence with distinct sub-populations of normal human mononuclear blood cells. The antibody T811 defines a surface antigen which is restricted to a subset of the T cell lineage. The antigen recognized by the second antibody, M522, is expressed on monocytes and polymorphonuclear leukocytes and, in addition, on 9-17% of nonadherent peripheral blood leukocytes (NAL). It is shown that the total NK activity of NAL is confined to the subset of cells expressing the M522-defined antigen. Moreover, the portion of NK cytotoxicity associated with T lymphocytes is mediated by a subpopulation which is characterized by the simultaneous expression of the T811- and the M522-defined antigens. This population comprises about 4% of NAL and could be isolated to a purity of greater than 85%.

Adherence of human lymphocytes to allogeneic tumour cell monolayers was found to depend on the presence of monocytes. Adherent lymphocytes could be separated from tumour cells by treatment with lidocaine followed by nylon wool passage. Tumour-adherent cells (70% E-RFC, 45% Fc gamma-R, 23% Fc mu-R, 5% monocytes) exhibited enriched natural killer (NK) activity not only against the tumour cell line used for isolation but also against seven other targets. When T cells were isolated subsequently as E-rosettes by density gradient centrifugation through Percoll, the enrichment in cytotoxicity was even more pronounced. Tumour-adherent T cells were severalfold enriched in both NK and antibody-dependent cell-mediated cytotoxicity (ADCC) activity. However, this enrichment was not paralleled by a concomitant increase in the number of T gamma cells (tumour-adherent T cells: 17% T gamma, 40% T mu ; tumour-nonadherent T cells: 12% T gamma, 60% T mu). Marked differences could be observed by staining with a monoclonal antibody that was raised against human leukaemic T gamma cells of high NK and ADCC activity. This antibody (T 8-11) stained 60% of tumour-adherent T cells, 20% of nonadherent T cells and 29% of T-cell controls. These results indicate that the spontaneous cytotoxic activity of human T cells resides within a small population, most of which are characterized by a specific surface antigen but not by conventional Fc gamma receptors.

Purified macrophage interleukin 1 (IL 1) induced a concentration-dependent inhibition of the proliferation of two commonly used tumor cell target lines, the human myeloid K562 and the murine T lymphoma Eb. In contrast, mastocytoma-derived P815 cells were not inhibited. The cytostatic action of IL 1 was not associated with direct cytotoxicity and was only partially reversible. PGE or interferon did not appear to mediate these effects. IL 1 treatment of the multipotential K562 cells revealed no morphologic evidence for the induction of specific differentiation. FACS analysis of IL 1-treated K562 cells showed a rapid decrease in transferrin receptor density, and a more delayed, but highly significant, increase in HLA-A,B,C antigen density. These findings provide one explanation for the frequently reported macrophage cytostatic actions against tumor cells, and indicate as well that IL 1, like interferon, may enhance the expression of Class I MHC antigens. These observations further extend the range of IL 1 actions and underscore the fundamental and direct role of this monokine in macrophage antitumor activity.