RESUMO
Paracoccidioidomycosis (PCM) is a systemic mycosis caused by the fungus Paracoccidioides brasiliensis (Pb). The cyclosporin A (CsA) is an immunosuppressant drug that inhibits calcineurin and has been described as a potential antifungal drug. The present study investigated the effect of CsA on the immune response, fungal load/antigenemia in experimental murine PCM. It was used four groups of BALB/c mice: (a) infected with 1 x 105 Pb18 yeast cells (Pb), (b) infected and treated with CsA every other day 10 mg/kg of CsA (s.c.) during 30 days (Pb/CsA), (c) treated with CsA (CsA) and (d) no infected/treated (PBS). The immune response was evaluated by lymphocyte proliferation, DTH assays to exoAgs, ELISA for IgG anti-gp43 (specific immune responses) and cytokine serum levels (IFN-γ, TNF-α, IL-4 and IL-10). Fungal load was determined by lung colony-forming units (CFU) counts, lung and liver histopathology analysis and antigenemia determined by inhibition-ELISA. As expected, CsA was able to inhibit the specific cellular and humoral immune response (P < 0.05), with decrease in serum IFN-γ, TNF-α and IL-4 levels (P < 0.05). Cyclosporin A treatment also resulted in significantly decreased lung Pb CFU (P < 0.05) as well as a lower number of yeasts in the lung and liver (P < 0.05) by histopathology. In concordance, the decreased antigenemia was observed in Pb/CsA group (P < 0.05). In conclusion, even with immunosuppressive action, treatment with CsA results in decreased lung fungal load/antigenemia in experimental PCM in BALB/c mice. Further study is required to determine whether this represents less severe disease or protection by CsA.
Assuntos
Ciclosporina/uso terapêutico , Pulmão/microbiologia , Paracoccidioides , Paracoccidioidomicose/tratamento farmacológico , Animais , Anticorpos Antifúngicos/sangue , Antígenos de Fungos/sangue , Antígenos de Fungos/imunologia , Contagem de Colônia Microbiana , Ciclosporina/imunologia , Ensaio de Imunoadsorção Enzimática , Proteínas Fúngicas/sangue , Proteínas Fúngicas/imunologia , Glicoproteínas/sangue , Glicoproteínas/imunologia , Hipersensibilidade Tardia/imunologia , Imunoglobulina G/sangue , Interferon gama/sangue , Interleucina-10/sangue , Interleucina-4/sangue , Fígado/microbiologia , Fígado/patologia , Pulmão/patologia , Ativação Linfocitária , Linfócitos/imunologia , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Paracoccidioides/efeitos dos fármacos , Paracoccidioides/crescimento & desenvolvimento , Paracoccidioides/imunologia , Paracoccidioidomicose/imunologia , Paracoccidioidomicose/microbiologia , Paracoccidioidomicose/patologia , Fator de Necrose Tumoral alfa/sangueRESUMO
The fungus Paracoccidioides brasiliensis is the pathogen of paracoccidioidomycosis (PCM), a systemic mycosis prevalent in Latin America. The loop-mediated isothermal amplification method (LAMP) was used in this study to detect the presence of P. brasiliensis in sputa samples from patients with chronic PCM, suspected PCM, and a negative control. The target P. brasiliensis gp43 gene was amplified in less than 4 hr in 11 of 18 sputa samples tested. The LAMP method had the advantage of speed and simplicity compared with the classic diagnostic methods such as the histopathological test or biological material culture and did not require sophisticated technical apparatus. It would be an important aid in cases where immediate treatment would mean patient survival, especially in immune-suppressed patients.
Assuntos
Antígenos de Fungos/genética , Proteínas Fúngicas/genética , Glicoproteínas/genética , Técnicas de Amplificação de Ácido Nucleico/métodos , Paracoccidioides/isolamento & purificação , Paracoccidioidomicose/diagnóstico , Escarro/microbiologia , Adulto , Idoso , Erros de Diagnóstico , Humanos , Masculino , Pessoa de Meia-Idade , Paracoccidioides/genética , Sensibilidade e EspecificidadeRESUMO
Infection is a major cause of morbidity and mortality in bone marrow transplant recipients and in patients with hematological malignancies. The source of infection is almost always endogenous flora or the hospital environment. The present study evaluated bone marrow transplant recipients and patients with hematological malignancies colonized and/or infected with filamentous fungi. During 1 year, environmental air samples were also taken from the bone marrow transplant unit by a modification of gravity air-setting plate (GASP) methodology. Fusarium spp. were the most prevalent genus in the fall and Cladosporium spp. in the winter. Clinically isolated strains grew better at 37 degrees C than environmental strains. According to NCCLS M-38P methods, environmental Aspergillus strains showed higher MICs to miconazol and itraconazol, and clinical Fusarium strains were less susceptible to fluconazole.
Assuntos
Microbiologia do Ar , Antifúngicos/farmacologia , Hospedeiro Imunocomprometido , Fungos Mitospóricos/efeitos dos fármacos , Fungos Mitospóricos/patogenicidade , Micoses/microbiologia , Aspergillus/efeitos dos fármacos , Aspergillus/isolamento & purificação , Aspergillus/patogenicidade , Transplante de Medula Óssea/efeitos adversos , Cladosporium/efeitos dos fármacos , Cladosporium/isolamento & purificação , Cladosporium/patogenicidade , Fusarium/efeitos dos fármacos , Fusarium/isolamento & purificação , Fusarium/patogenicidade , Neoplasias Hematológicas/complicações , Hospitais de Ensino , Humanos , Testes de Sensibilidade Microbiana , Fungos Mitospóricos/classificação , Fungos Mitospóricos/isolamento & purificaçãoRESUMO
Cryptococcus neoformans is an important fungal pathogen in immunocompromised hosts. Capsulation, urease and melanin synthesis activity of the fungus are well known virulence factors. Although artificial melanin-deficient mutants of Cr. neoformans have been investigated, the clinical mutant is rare. We found a Cr. neoformans isolate in the cerebrospinal fluid of an AIDS patient which produced a light tan colony on a caffeic acid cornmeal agar (CACA) plate. The mycological feature of the isolate was as follows; normal capsulation, defective inositol assimilation ability, serotype A; urease-positive; mating type alfa; haploid; extremely slow growth in RPMI 1640 medium, Sabouraud dextrose broth, brain heart infusion broth and yeast nitrogen base; lower production of melanin with L-DOPA substrate; and low virulence to ddY mice. We also investigated the partial DNA sequence of CNLAC1 gene between the 3085th to 3623rd base. There were many substitutions, 3 insertions and 3 deletions in the isolate compared with GenBank accession number L22866. The result indicated some functional disorder in the gene. Although the CACA plate is an excellent selective medium for Cr. neoformans, other identification methods should also be used.
Assuntos
Síndrome da Imunodeficiência Adquirida/microbiologia , Cryptococcus neoformans/isolamento & purificação , Adulto , Cryptococcus neoformans/genética , Feminino , Humanos , Hospedeiro Imunocomprometido , Técnicas MicrobiológicasRESUMO
Candida dubliniensis is a newly-recognized Candida species and an important infectious pathogen, particularly for HIV-positive patients. >From oral smear samples from the radix linguae of 173 HIV-positive children, we obtained four yeast isolates which took a blue-green color on CHROMagar Candida plate at 37 degrees C for 48 hours from one HIV-positive 3-year-old boy in Brazil. The isolates were difficult to grow on potato dextrose agar plate at 42 degrees C, produced abundant chlamydospores on a cornmeal agar plate with Tween 80, and sprouted germ tubes in saline with horse serum, and the antigenic profile by CANDIDA CHECK test was useless. Carbohydrate assimilation tests by ID32C showed no reference code number in the reference book. The isolates were subjected to molecular biological assay of the DNA sequence of the large-subunit ribosomal DNA region (D1/D2) and randomly amplified polymorphic DNA (RAPD). The DNA sequence agreed with those of standard C. dubliniensis strains, and therefore, the isolates were identified as C. dubliniensis. RAPD band pattern analysis indicated that the clinical isolates might summarize one genotype. Although the child did not present oral lesions, the fungus might be latent for opportunistic infection.