IFN-γ Production Depends on IL-12 and IL-18 Combined Action and Mediates Host Resistance to Dengue Virus Infection in a Nitric Oxide-Dependent Manner

Abstract

Dengue is a mosquito-borne disease caused by one of four serotypes of Dengue virus (DENV-1–4). Severe dengue infection in humans is characterized by thrombocytopenia, increased vascular permeability, hemorrhage and shock. However, there is little information about host response to DENV infection. Here, mechanisms accounting for IFN-γ production and effector function during dengue disease were investigated in a murine model of DENV-2 infection. IFN-γ expression was greatly increased after infection of mice and its production was preceded by increase in IL-12 and IL-18 levels. In IFN-γ^−/− mice, DENV-2-associated lethality, viral loads, thrombocytopenia, hemoconcentration, and liver injury were enhanced, when compared with wild type-infected mice. IL-12p40^−/− and IL-18^−/− infected-mice showed decreased IFN-γ production, which was accompanied by increased disease severity, higher viral loads and enhanced lethality. Blockade of IL-18 in infected IL-12p40^−/− mice resulted in complete inhibition of IFN-γ production, greater DENV-2 replication, and enhanced disease manifestation, resembling the response seen in DENV-2-infected IFN-γ^−/− mice. Reduced IFN-γ production was associated with diminished Nitric Oxide-synthase 2 (NOS2) expression and NOS2^−/− mice had elevated lethality, more severe disease evolution and increased viral load after DENV-2 infection. Therefore, IL-12/IL-18-induced IFN-γ production and consequent NOS2 induction are of major importance to host resistance against DENV infection. Dengue fever and its severe forms, dengue hemorrhagic fever and dengue shock syndrome, are the most prevalent mosquito-borne diseases on Earth. It is caused by one of four serotypes of Dengue virus (DENV-1–4). At present, there are no vaccines or specific therapies for dengue and treatment is supportive. Host response to infection is also poorly understood. Here, using a DENV-2 strain that causes a disease that resembles the severe manifestations of Dengue in humans, we demonstrate that IFN-γ production is essential for the host to deal with infection. We have also shown that IFN-γ production during DENV infection is controlled by the cytokines IL-12 and IL-18. Finally, we show that one of the mechanisms triggered by IFN-γ during host response to DENV infection is the production of Nitric Oxide, an important virustatic metabolite. Mice deficient for each of these molecules present marked increase in DENV replication after infection and more severe disease. Altogether, this study demonstrates that the IL-12/IL-18-IFN-γ-NO axis plays a major role in host ability to deal with primary DENV infection. These data bear relevance to the understanding of antiviral immune responses during Dengue disease and may aid in the rational design of vaccines against DENV infection.