Detrimental Effect of Fungal 60-kDa Heat Shock Protein on Experimental Paracoccidioides brasiliensis Infection

Fabrício Freitas Fernandes; Leandro Licursi de Oliveira; Taise Natali Landgraf; Gabriela Peron; Marcelo Vieira Costa; Arlete A M Coelho-Castelo; Vânia L D Bonato; Maria-Cristina Roque-Barreira; Ademilson Panunto-Castelo

doi:10.1371/journal.pone.0162486

Detrimental Effect of Fungal 60-kDa Heat Shock Protein on Experimental Paracoccidioides brasiliensis Infection

PLoS One. 2016 Sep 6;11(9):e0162486. doi: 10.1371/journal.pone.0162486. eCollection 2016.

Authors

Fabrício Freitas Fernandes¹, Leandro Licursi de Oliveira², Taise Natali Landgraf³, Gabriela Peron⁴, Marcelo Vieira Costa⁵, Arlete A M Coelho-Castelo³, Vânia L D Bonato³, Maria-Cristina Roque-Barreira¹, Ademilson Panunto-Castelo⁵

Affiliations

¹ Department of Cellular and Molecular Biology, Ribeirão Preto School of Medicine, University of São Paulo, Ribeirão Preto, SP, Brazil.
² Department of General Biology, Federal University of Viçosa, Viçosa, MG, Brazil.
³ Department of Biochemistry and Immunology, School of Medicine of Ribeirão Preto, University of São Paulo, Ribeirão Preto, SP, Brazil.
⁴ Department of Structural and Functional Biology, Institute of Biology, State University of Campinas, Campinas, SP, Brazil.
⁵ Department of Biology, Ribeirão Preto Faculty of Philosophy, Sciences and Letters, University of São Paulo, Ribeirão Preto, SP, Brazil.

Abstract

The genus Paracoccidioides comprises species of dimorphic fungi that cause paracoccidioidomycosis (PCM), a systemic disease prevalent in Latin America. Here, we investigated whether administration of native 60-kDa heat shock protein of P. brasiliensis (nPbHsp60) or its recombinant counterpart (rPbHsp60) affected the course of experimental PCM. Mice were subcutaneously injected with nPbHsp60 or rPbHsp60 emulsified in complete's Freund Adjuvant (CFA) at three weeks after intravenous injection of P. brasiliensis yeasts. Infected control mice were injected with CFA or isotonic saline solution alone. Thirty days after the nPbHsp60 or rPbHsp60 administration, mice showed remarkably increased fungal load, tissue inflammation, and granulomas in the lungs, liver, and spleen compared with control mice. Further, rPbHsp60 treatment (i) decreased the known protective effect of CFA against PCM and (ii) increased the concentrations of IL-17, TNF-α, IL-12, IFN-γ, IL-4, IL-10, and TGF-β in the lungs. Together, our results indicated that PbHsp60 induced a harmful immune response, exacerbated inflammation, and promoted fungal dissemination. Therefore, we propose that PbHsp60 contributes to the fungal pathogenesis.

MeSH terms

Animals
Antigens, Fungal / administration & dosage*
Chaperonin 60 / administration & dosage*
Disease Progression
Freund's Adjuvant / administration & dosage
Fungal Proteins / administration & dosage*
Gene Expression
Interferon-gamma / genetics
Interferon-gamma / immunology
Interleukins / genetics
Interleukins / immunology
Liver / drug effects
Liver / immunology
Liver / microbiology
Liver / pathology
Lung / drug effects
Lung / immunology
Lung / microbiology
Lung / pathology
Male
Mice
Mice, Inbred BALB C
Paracoccidioides / growth & development
Paracoccidioides / pathogenicity*
Paracoccidioidomycosis / immunology
Paracoccidioidomycosis / microbiology
Paracoccidioidomycosis / pathology*
Recombinant Proteins / administration & dosage
Spleen / drug effects
Spleen / immunology
Spleen / microbiology
Spleen / pathology
Tumor Necrosis Factor-alpha / genetics
Tumor Necrosis Factor-alpha / immunology

Substances

Antigens, Fungal
Chaperonin 60
Fungal Proteins
Interleukins
Recombinant Proteins
Tumor Necrosis Factor-alpha
Interferon-gamma
Freund's Adjuvant

Grants and funding

APC (Grant #2009/14777-1, and #2013/12278-3), FFF (# 2009/03235-3), and TNL (# 2012/08552-0) were supported by grants from Fundação de Amparo à Pesquisa do Estado de São Paulo. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.