Abstract
Paracoccidioidomycosis (PCM) is one of the most frequent systemic mycoses in Latin America. It affects mainly male rural workers in impoverished regions, and the therapy can last up to two years or use drugs that are very toxic. Given the need for novel safe and effective approaches to treat PCM, we have been developing monoclonal antibodies (mAbs) that could be used not only to block specific fungal targets, but also modulate the host’s antifungal immunity. In this work we show the generation of and promising results with an mAb against Heat Shock Protein (HSP)90, a molecular chaperone that is an important virulence factor in fungi. Using recombinant Paracoccidioides lutzii (Pb01) and P. brasiliensis (Pb18) HSP90 proteins produced in E. coli, we immunized mice and generated polyclonal antibodies and an IgG1 hybridoma mAb. The proteins were very immunogenic and both the polyclonal serum and mAb were used in immunofluorescence experiments, which showed binding of antibodies to the yeast cell surface. The mAb successfully opsonized P. lutzii and P. brasiliensis cells in co-incubations with J774.16 macrophage-like cells. Our results suggest that this mAb could serve as the basis for new immunotherapy regimens for PCM.
Highlights
Paracoccidioidomycosis (PCM) is an invasive fungal disease endemic to Latin America
The extra bands observed in stained SDS-PAGE gels were not recognized by the anti-His antibody, indicating they are either HSP90 degradation products that do not contain the his-tag or unrelated proteins (Figure 2)
Sera from mice that were immunized with P. brasiliensis or P. lutzii recombinant HSP90 were able to recognize and bind HSP90, as were sera from animals that were immunized with a mixture of both proteins
Summary
Paracoccidioidomycosis (PCM) is an invasive fungal disease endemic to Latin America. It affects mostly poor rural workers and has a high medical and social impact due to its chronicity and frequent fibrotic sequelae, which lead to inability to work and poor quality of life [1]. J. Fungi 2020, 6, 193; doi:10.3390/jof6040193 www.mdpi.com/journal/jof. J. Fungi 2020, 6, 193 treated and cured, the available therapeutic tools are far from optimal. The drug of choice for severe cases, Amphotericin B (AmB), is very nephrotoxic [2] and requires hospitalization for intravenous use
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