Nitric oxide mediation of active immunosuppression associated with graft-versus-host reaction

In the immunosuppression accompanying the lethal systemic graft-versus-host reaction (GVHR) directed against minor histocompatibility antigens in irra diated adult mice, we previously determined that non-T, non-B, L-leucine me thyl ester (LME)-sensitive cells were implicated via two different mechanis ms: one, which is interferon-gamma (IFN-gamma)-dependent and affects both T -cell proliferative responses and thymus-independent antibody production by CD5(+) B cells; and a second, which is IFN-gamma-independent and affects B -cell proliferative responses. Because IFN-gamma induces the production of nitric oxide (NO), a potent immunosuppressive molecule, we investigated the involvement of NO in the suppression mediated by the LME-sensitive cells. Inducible NO synthase (iNOS) mRNA, iNOS protein, and the stable end product s of iNOS pathway, L-citrulline and nitrite, were detected early in GVHR in LME-sensitive spleen cells taken ex vivo and could be amplified in vitro b y T and B mitogens. Inhibition of NO production with arginine analogs (amin oguanidine, N-G-monomethyl-L-arginine [LMMA]), like anti-IFN-gamma antibodi es, reversed suppression of both T-cell responses to concanavalin A and CD5 (+) B-cell responses, but not of B-cell response to lipopolysaccharides (LP S). The GVHR-associated, IFN-gamma-dependent immunosuppression of T-cell pr oliferation and of antibody synthesis by CD5(+) B cells is the consequence of NO production by LME-sensitive cells. Immunohistochemical analyses indic ate that these cells belong to the macrophage lineage. (C) 1999 by The Amer ican Society of Hematology.