title: Additive effect of rPb27 immunization and chemotherapy in experimental paracoccidioidomycosis.
abstract: Paracoccidioidomycosis, PCM, the major systemic mycosis in Latin America, is caused by the termally dimorphic fungus Paracoccidioides brasiliensis and requires extended periods of chemotherapy with a significant frequency of relapsing disease. The search for new alternatives of treatment is necessary. rPb27 is an antigenic protein from P. brasiliensis that already showed a significant protective activity as a vaccine for PCM in experimental models. The cDNA of rPb27 was subcloned into a pET-DEST 42 plasmid, expressed in E. coli with a his-tag and purified by affinity chromatography. Immunization with this recombinant protein and chemotherapy were used together in an attempt to improve treatment of PCM. For this, BALB/c mice were challenged with pathogenic P. brasiliensis strain and after immunized with rPb27, in the presence of Corynebacterium parvum and Al(OH)(3), some groups were also treated with fluconazole. After 40 days of treatment, the combined drug/rPb27 administration controlled PCM in the liver and spleen, with long lasting protection, and largely preserved tissues structures of these organs. Additionally, in the lungs after 40 days of treatment there was a significant reduction in the fungal load and size of lesions. At the same time, the levels of TNF-α were higher than infected-only mice. Moreover, significant levels of anti-rPb27 specific IgG1, IgG2a and IgG2b isotypes were detected in the sera of mice immunized with rPb27 fluconazole treated or not. These results showed an additive protective effect of rPb27 immunization and chemotherapy, suggesting that an rPb27-based vaccine can be used to enhance PCM antifungal treatment.
authors.affiliation: Departamento de Bioquímica e Imunologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte, Brazil. cfernandes.viviane@gmail.com
authors.forename: Viviane C
authors.initials: VC
authors.lastname: Fernandes
authors.forename: Estefânia M N
authors.initials: EM
authors.lastname: Martins
authors.forename: Jankerle N
authors.initials: JN
authors.lastname: Boeloni
authors.forename: Juliana B
authors.initials: JB
authors.lastname: Coitinho
authors.forename: Rogéria
authors.initials: R
authors.lastname: Serakides
authors.forename: Alfredo M
authors.initials: AM
authors.lastname: Goes
mesh_terms: D000818:Animals; D000935:Antifungal Agents / Q000494:pharmacology / Q000627:therapeutic use*; D000946:Antigens, Fungal / Q000276:immunology*; D003001:Cloning, Molecular; D003114:Colony-Forming Units Assay; D015725:Fluconazole / Q000494:pharmacology / Q000627:therapeutic use; D005657:Fungal Vaccines / Q000276:immunology*; D007114:Immunization; D007074:Immunoglobulin G / Q000096:biosynthesis / Q000276:immunology; D008099:Liver / Q000187:drug effects / Q000382:microbiology / Q000473:pathology; D008168:Lung / Q000187:drug effects / Q000382:microbiology / Q000473:pathology; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D009928:Organ Specificity / Q000187:drug effects; D010229:Paracoccidioidomycosis / Q000188:drug therapy* / Q000276:immunology* / Q000382:microbiology; D011994:Recombinant Proteins / Q000276:immunology / Q000302:isolation & purification; D013154:Spleen / Q000187:drug effects / Q000382:microbiology / Q000473:pathology; D014132:Trachea / Q000187:drug effects / Q000382:microbiology / Q000473:pathology
chemical_list: D000935:Antifungal Agents; D000946:Antigens, Fungal; D005657:Fungal Vaccines; D007074:Immunoglobulin G; D011994:Recombinant Proteins; D015725:Fluconazole