Tolerogenic Plasmacytoid Dendritic Cells Control Paracoccidioides brasiliensis Infection by Inducting Regulatory T Cells in an IDO-Dependent Manner

Abstract

Plasmacytoid dendritic cells (pDCs), considered critical for immunity against viruses, were recently associated with defense mechanisms against fungal infections. However, the immunomodulatory function of pDCs in pulmonary paracoccidiodomycosis (PCM), an endemic fungal infection of Latin America, has been poorly defined. Here, we investigated the role of pDCs in the pathogenesis of PCM caused by the infection of 129Sv mice with 1 x 10 ⁶ P. brasiliensis-yeasts. In vitro experiments showed that P. brasiliensis infection induces the maturation of pDCs and elevated synthesis of TNF-α and IFN-β. The in vivo infection caused a significant influx of pDCs to the lungs and increased levels of pulmonary type I IFN. Depletion of pDCs by a specific monoclonal antibody resulted in a less severe infection, reduced tissue pathology and increased survival time of infected mice. An increased influx of macrophages and neutrophils and elevated presence of CD4 ⁺ and CD8 ⁺ T lymphocytes expressing IFN-γ and IL-17 in the lungs of pDC-depleted mice were also observed. These findings were concomitant with decreased frequency of Treg cells and reduced levels of immunoregulatory cytokines such as IL-10, TGF-β, IL-27 and IL-35. Importantly, P. brasilienis infection increased the numbers of pulmonary pDCs expressing indoleamine 2,3-dioxygenase-1 (IDO), an enzyme with immunoregulatory properties, that were reduced following pDC depletion. In agreement, an increased immunogenic activity of infected pDCs was observed when IDO-deficient or IDO-inhibited pDCs were employed in co-cultures with lymphocytes Altogether, our results suggest that in pulmonary PCM pDCs exert a tolerogenic function by an IDO-mediated mechanism that increases Treg activity.

Author Summary

The fungus Paracoccidioides brasiliensis causes paracoccidioidomycosis (PCM), the most relevant deep mycosis in Latin America. The plasmacytoid dendritic cells (pDCs) are important immune cells involved in protection against viral infections, but their role in fungal infections remains unclear. Here, we investigated the role of pDCs in the pathogenesis of pulmonary PCM using a monoclonal antibody to deplete this DC subset. pDCs depletion leads to a less severe PCM associated with increased T cell response mainly mediated by Th1 and Th17 cells. The lung homogenates of depleted mice showed diminished levels of type I IFN and anti-inflammatory cytokines. In addition, a reduced number of regulatory T cells (Treg) paralleled a diminished number pDCs expressing IDO, a potent immunoregulatory enzyme. In agreement, pDCs of IDO ^-/- mice or IDO-inhibited pDCs stimulated by P. brasiliensis yeasts expanded elevated numbers of T cells concomitant with a reduced expansion of Treg cells. Taken together, our results demonstrate a tolerogenic activity of pDCs that enhances the severity of a pulmonary mycosis mediated by the concerted action of IDO and Treg cells. These results reveal a new function for pDCs in primary fungal infections and open new perspectives for immunotherapeutic procedures of PCM involving the control of IDO and Treg activity.