Genetic characterization of morphologically variant strains of Paracoccidioides brasiliensis

Molecular characterization of Paracoccidioides brasiliensis variant strains that had been preserved under mineral oil for decades was carried out by random amplified polymorphic DNA analysis (RAPD). On P. brasiliensis variants in the transitional phase and strains with typical morphology, RAPD produced reproducible polymorphic amplification products that differentiated them. A dendrogram based on the generated RAPD patterns placed the 14 P. brasiliensis strains into five groups with similarity coefficients of 72%. A high correlation between the genotypic and phenotypic characteristics of the strains was observed. A 750 bp-RAPD fragment found only in the wild-type phenotype strains was cloned and sequenced. Genetic similarity analysis using BLASTx suggested that this RAPD marker represents a putative domain of a hypothetical flavin-binding monooxygenase (FMO)-like protein of Neurospora crassa.FiocruzBritish Council Progra

Gelatinase activity of exoantigens from virulent and non-virulent isolates of Paracoccidioides brasiliensis

Differences in the occurrence of components with gelatinase activity were detected among four isolates of Paracoccidioides brasiliensis: Pb339 and Pb18 (highly virulent), and Pb265 and Pb18AV (very low virulence). Culture filtrates from these isolates were electrophoresed in substrate gels and tested for gelatinase activity. Pb339 showed three enzyme bands of apparent molecular masses: 43, 53 and 78 kDa; Pb18 had two bands, one at 59 kDa and another with molecular mass higher than 78 kDa. Isolate Pb18AV showed only one band at 78 kDa and Pb265 exhibited a component of molecular mass which failed to enter the separating gel.CENT PUBL HLTH LAB,MYCOL REFERENCE LAB,LONDON,ENGLANDESCOLA PAULISTA MED,DEPT MOLEC BIOL,SAO PAULO,SP,BRAZILESCOLA PAULISTA MED,DEPT MOLEC BIOL,SAO PAULO,SP,BRAZILWeb of Scienc

Interleukin-12 protects mice against disseminated infection caused by Paracoccidioides brasiliensis but enhances pulmonary inflammation

Paracoccidioides brasiliensis is a facultative, intracellular pathogen causing the most important deep mycosis in Latin America. As the production of IFN-gamma and induction of cell-mediated immunity to P. brasiliensis is of critical importance in host defense, the immunotherapeutic effect of exogenous IL-12 administration was studied in a murine model of susceptibility to pulmonary infection. rIL-12 treatment led to a less disseminated disease, as confirmed by decreased fungal loads in liver and spleen. Administration of rIL-12 did not affect fungal growth in the lungs, although it did induce an augmented pulmonary mononuclear cell inflammation. IL-12 treatment induced an early (week 1) increase in pulmonary IFN-gamma, but decreased cytokine and specific antibody (IgG1 and IgG3) production at week 8 after infection. These results show that IL-12 administration induces a less severe infection, but the high inflammatory response detected in the lungs precludes its possible use as a new therapeutic tool for severe paracoccidioidomycosis. (C) 2002 Elsevier Science (USA).Univ São Paulo, Dept Imunol, Inst Ciencias Biomed, BR-05508900 São Paulo, BrazilUniversidade Federal de São Paulo, Dept Patol, Fac Med, São Paulo, BrazilInsat Adolpho Lutz, São Paulo, BrazilUniversidade Federal de São Paulo, Dept Patol, Fac Med, São Paulo, BrazilWeb of Scienc