B-chronic lymphocytic leukemia cells exert an in vitro cytotoxicity mediated by tumor necrosis factor α

Tumor necrosis factor a (TNF alpha) is constitutively produced by B-chronic lymphocytic leukemia (B-CLL) cells and may act as an autocrine factor for their growth and survival. However, very few data are available on the possible cytotoxic effect of TNF alpha produced by B-CLL cells. This study investigated whether B-CLL cells exert in vitro cytotoxicity by TNF alpha and if so, whether this cytotoxicity can be modulated by cytokines. In 8 of 12 patients (66.6%), B-CLL cells in vitro constitutively produced TNF alpha and exerted a TNF alpha-mediated cytotoxicity, evaluated in an 18-h(51)r release assay, against the TNF alpha-sensitive Jurkat, U937 and K562 cell lines but not against the TNF alpha-resistant Raji cell line. Involvement of TNF alpha in B-CLL cell cytotoxicity is demonstrated by the fact that anti-TNF alpha antibodies strongly inhibited it and supernatants of cytotoxic cultures contained TNF alpha and mediated a completely TNF alpha-dependent cytotoxicity. When the cytotoxic B-CLL cells were stimulated with interleukin (IL)-2 plus IL-12, there was increased TNF alpha mRNA expression, TNFa production and TNF alpha-mediated cytotoxicity. All eight patients with cytotoxic leukemic cells had progressive disease and six of these also expressed high levels of ZAP-70 protein. In the other four patients (33.3%), B-CLL cells did not produce TNF alpha in vitro and were not cytotoxic, either spontaneously or after IL-2 plus IL-12 stimulation. Of these four patients, three had stable disease and one had progressive disease. The patient with progressive disease and one of the three with stable disease expressed low levels of ZAP-70 protein. We conclude that a group of B-CLL patients with progressive disease have leukemic B cells able to exert in vitro a TNFa-mediated cytotoxicity, which is modulated by cytokines. (c) 2005 Elsevier Ltd. All rights reserved.