Interleukin-2-induced nitric oxide synthase and nuclear factor-kappaB activity in activated natural killer cells and the production of interferon-gamma.

We have previously shown that inducible nitric oxide synthase (iNOS) was up-regulated in natural killer (NK) cells when AK-5 tumour cells were transplanted subcutaneously into syngeneic Wistar rats. This study was designed to investigate the role of interleukin (IL)-2 during the induction of iNOS and to understand the subsequent events involved in NK cell activation. There was up-regulation of iNOS expression when naïve NK cells were cultured in the presence of recombinant IL-2. These NK cells produced a higher nitrite content and possessed cytotoxic activity against YAC-1 and AK-5 tumour cells. Induction of iNOS enhanced nuclear factor (NF)-kappaB binding activity in IL-2 activated NK cells, which was confirmed using L-NAME, an NO synthesis inhibitor. Addition of L-NAME along with rIL-2 significantly blocked NF-kappaB activity and also down-regulated the production of NO and the cytotoxic activity of NK cells. Furthermore, injection of anti-IL-2 antibody in subcutaneous tumour transplanted animals abrogated significantly the expression of iNOS and NF-kappaB activity, leading to reduced NO production and cytotoxic activity of NK cells against YAC-1 and AK-5 cells. In addition, the expression of interferon (IFN)-gamma by NK cells was also inhibited in anti-IL-2 antibody injected animals compared with the control animals. Finally, there was enhanced tumour growth and delayed regression in anti-IL-2 injected animals compared with control animals.