Nitric oxide production by mouse bone marrow-derived dendritic cells: implications for the regulation of allogeneic T cell responses.

Abstract

Dendritic cells (DC) are the most potent known antigen presenting cells, and play important roles both in immunity and tolerance induction. Nitric oxide (NO) is an important effector molecule that is involved in numerous aspects of the immune response. There have been no accounts to date of efforts to determine NO generation by well-characterized DC. In this report we describe the production of NO by highly purified DEC 205+ DC propagated from mouse bone marrow in response to granulocyte/macrophage-colony stimulating factor (GM-CSF) + interleukin-4 (IL-4). NO synthesis was induced in DC by interferon-gamma (IFN-gamma) and lipopolysaccharide (LPS), and was blocked by the inhibitor of nitric oxide synthase (NOS), NG-monomethyl-L-arginine (NMMA). Both "mature" B7-2+ (CD86+) DC and B7-2- (CD86-) DC progenitors could be induced to release NO. NO was also recovered from the supernatants of primary mixed leukocyte cultures containing comparatively high concentrations of B7-2+ DC in relation to purified allogeneic T cells. Furthermore, inhibition of NO release in these cultures by NMMA resulted in an increase in T cell proliferation. These observations suggest that NO may be an important soluble mediator of the interaction between DC and activated T cells. In addition to its ability to inhibit T cell proliferation, NO was also shown to induce programmed cell death in DC. This was visualized by the detection of DNA strand breaks with in situ nick translation. The percentage of DC apoptosis correlated with the level of NO in the cultures. Apoptosis was inhibited by the addition of NMMA. These results indicate that DC have the capacity both to stimulate and potentially limit the same allogeneic T cell response, in accordance with their production of NO.