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Regulation of Large Granular Lymphocytes and Human T Cell Growth and Function by Recombinant Interleukin 2. II. Acquisition of Potent Cytotoxic Capabilities

Human large granular lymphocytes (LGL) and T cells were separated from peripheral blood on discontinuous density gradients of Percoll. On a per cell basis, cultured LGL demonstrated higher levels of cytotoxicity against K562 cells than fresh LGL. Cultured T cells acquired cytotoxicity against K562 cells, although they were less cytotoxic than fresh or cultured LGL. During culture, these LGL retained the 3G8, NKH1, OKM1, and OKT10 antigens. Cultured T cells retained the T101 antigen and acquired the OKM1 and OKT10 antigens, but remained negative for the 3G8 or NKH1 antigens. A series of pharmacologic agents known to inhibit cytotoxicity of fresh LGL were also tested for their effects on cultured LGL and T cells. Ethylenediaminetetraacetic acid, known to inhibit the binding of effectors to target cells, inhibited the cytotoxicity of cultured LGL and T cells to the same degree that it inhibited the cytotoxicity of fresh LGL. In contrast, agents which primarily effect post‐binding events in cytotoxicity (trypsin, D‐mannose 6‐P, dexamethasone, prostaglandin E2, anti‐LGL granule antibody, and 9.1C3 monoclonal antibody) inhibited the cytotoxicity of cultured LGL and T cells to a much lesser extent than fresh LGL. In addition, on the basis of Michaelis‐Menten's kinetic analysis, cultured LGL and T cells developed a higher binding affinity to K562 cells than fresh LGL.