Augmentation of anti-cytokine immunotherapy by combining neutralizing monoclonal antibodies to interferon-γ and the interferon-γ receptor: protection in endotoxin shock

Abstract

We have employed neutralizing monoclonal antibodies to mouse interferon-γ (IFNγ) and to the receptor for mouse IFNy in studies designed to assess the protective efficacy of each of these monoclonals, administered either separately or in combination, in endotoxin-induced lethality. While pretreatment with either antibody alone, at doses of 200 μg per mouse, provided limited protection (70-50% lethality) in comparison to non-neutralizing antibody controls (100% lethality), the two monoclonal antibodies administered together provided a substantially greater level of protection (17% lethality). Although administration of 100 μg per mouse of either monoclonal alone was not protective (more than 65% lethality), a combination of both antibodies at this dose provided significant protection (19% lethality). In addition, administration of both antibodies 30 min post-endotoxin challenge also demonstrated significant protection in comparison to single antibody immunotherapy. In vitro studies using mouse peritoneal macrophages stimulated with LPS and IFNy have established confirmatory data for a synergistic effect of neutralizing antibody to IFNy and the IFNγ receptor in inhibiting macrophage activation as assessed by production of nitric oxide. These results provide a strong rationale for dual targeting of ligand and receptor in single cytokine immunotherapy.