Patogenicidade e imunogenicidade de isolados clínicos do complexo Paracoccidioides brasiliensis
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2019-02-21
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Universidade Estadual Paulista (Unesp)
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Introdução. A correlação entre gravidade da paracoccidioidomicose e patogenicidade e imunogenicidade dos fungos causadores tem sido pouco investigada e foi o objetivo deste estudo. Metodologia. As cincos cepas Pb192, Pb234, Pb326, Pb417 e Pb531 foram identificadas pelo seqüenciamento da região Exon 2 da gp43. A patogenicidade foi determinada pelo cálculo da dose letal 50% (DL50%) e pela contagem do número de unidades formadoras de colônias, realizada na sexta semana pós-infecção de camundongos BALB/c. A imunogenicidade foi determinada pela avaliação da resposta imune humoral específica, utilizando-se a reação de imunodifusão dupla em gel de ágar e da imunidade celular, determinada pela concentração das citocinas interleucina -2, interleucina-10, interferon-γ, fator de necrose tumoral – α e do fator de crescimento do endotélio vascular, em tecido pulmonar. Quatro amostras clínicas foram recém-isoladas de pacientes com paracoccidioidomicose, provenientes da Região de Botucatu - os isolados Pb234 e Pb417, de pacientes com a forma crônica moderada; o Pb326 de um caso com a forma aguda grave; e o Pb531, de um caso com a forma crônica grave. As demais cepas Pb192, Pb01 e 8334 foram cedidas pelo laboratório de Moléstias infecciosas. Resultados. As cepas Pb417 e Pb326 agruparam-se às cepas identificadas como P. brasiliensis S1a, a Pb531 às P. brasiliensis S1b e as cepas Pb234 e Pb192 às cepas depositadas como P. restrepiensis (PS3). Os resultados demonstraram correlação direta entre patogenicidade do isolado e gravidade da doença. Os menores valores de DL50% foram observados em isolados dos casos graves da doença, Pb531 e Pb326. A cepa Pb531 foi considerada de alta virulência, Pb326 de virulência intermediaria e os outros isolados Pb192, Pb234, Pb417, Pb01, 8334 de baixa virulência. Os níveis séricos de anticorpos só foram detectadas nos animais infectados com a cepa Pb326 na sexta semana pós-infecção. Nos dois primeiros estágios, 2ª e 4ª semana de infecção, observou-se uma distribuição razoavelmente uniforme da concentração pulmonar de citocinas e do fator de crescimento do endotélio vascular; porem, a 6ª semana observou-se evidente predomínio da interleucina-10, com pequenas variações entre isolados. Conclusões. Observou-se correlação direta entre patogenicidade do isolado e gravidade do paciente e grande semelhança de imunogenicidade dos isolados; P. restrepiensis também se encontra na Região de Botucatu (Brasil) e não apenas na Colômbia.
Introduction. The investigation of paracoccidioidomycosis and pathogenicity and immunogenicity of the provoking fungi has been little investigated and was the objective of this study. Methodology. As strains, Pb192, Pb234, Pb326, Pb417 and Pb531 were included by sequencing the Exon 2 region of gp43. The pathogenicity was determined by calculating the 50% lethal dose (LD50%) and by counting the number of colony forming units performed in the sixth week post-infection of BALB / c mice. Immunogenicity was determined by the humoral immune response, using the agar gel immunodiffusion reaction and the cellular immunity, determined the concentration of cytokines interleukin-2, interleukin-10, interferonγ, tumor necrosis factor - and factor of vascular endothelial growth in lung tissue. Surgical has been associated with patients with paracoccidioidomycosis, derived from the Botucatu - the Pb234 and Pb417; the Pb326 of a case with a severe severe form; and Pb531, of a case with severe chronic form. The other strains Pb192, Pb01 and 8334 were transferred by the laboratory of Infectious Diseases. Results. Pb417 and Pb326 strains were grouped into the associated strains P. brasiliensis S1a, Pb531 to P. brasiliensis S1b and strains Pb234 and Pb192 to strains deposited as P. restrepiensis (PS3). The results demonstrate the pathogenicity of disease error and severity. The small LD 50 values were measured in the following cases: severe disease, Pb531 and Pb326. Pb531 strain was considered to be highly virulent, intermediate virulence Pb326 and other low virulence Pb192, Pb234, Pb417, Pb01, 8334. Serum antibodies were detected in animals infected with the Pb326 strain at the sixth postinfection week. In the first two stages, 2nd and 4th week of infection, we observed the presence of a coronary lymph node and a vascular endocrine growth factor; However, the 6th week observed a predominance of interleukin-10, with small averages between the results. Observing the pathogenicity of the weight and the severity of the immunogenicity of the isolates; P. restrepiensis is also found in the Botucatu Region (Brazil) and not only in Colombia.
Introduction. The investigation of paracoccidioidomycosis and pathogenicity and immunogenicity of the provoking fungi has been little investigated and was the objective of this study. Methodology. As strains, Pb192, Pb234, Pb326, Pb417 and Pb531 were included by sequencing the Exon 2 region of gp43. The pathogenicity was determined by calculating the 50% lethal dose (LD50%) and by counting the number of colony forming units performed in the sixth week post-infection of BALB / c mice. Immunogenicity was determined by the humoral immune response, using the agar gel immunodiffusion reaction and the cellular immunity, determined the concentration of cytokines interleukin-2, interleukin-10, interferonγ, tumor necrosis factor - and factor of vascular endothelial growth in lung tissue. Surgical has been associated with patients with paracoccidioidomycosis, derived from the Botucatu - the Pb234 and Pb417; the Pb326 of a case with a severe severe form; and Pb531, of a case with severe chronic form. The other strains Pb192, Pb01 and 8334 were transferred by the laboratory of Infectious Diseases. Results. Pb417 and Pb326 strains were grouped into the associated strains P. brasiliensis S1a, Pb531 to P. brasiliensis S1b and strains Pb234 and Pb192 to strains deposited as P. restrepiensis (PS3). The results demonstrate the pathogenicity of disease error and severity. The small LD 50 values were measured in the following cases: severe disease, Pb531 and Pb326. Pb531 strain was considered to be highly virulent, intermediate virulence Pb326 and other low virulence Pb192, Pb234, Pb417, Pb01, 8334. Serum antibodies were detected in animals infected with the Pb326 strain at the sixth postinfection week. In the first two stages, 2nd and 4th week of infection, we observed the presence of a coronary lymph node and a vascular endocrine growth factor; However, the 6th week observed a predominance of interleukin-10, with small averages between the results. Observing the pathogenicity of the weight and the severity of the immunogenicity of the isolates; P. restrepiensis is also found in the Botucatu Region (Brazil) and not only in Colombia.
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Paracoccidioidomycosis, Systemic mycoses, Virulence, Severity, Murine model and sequencing, Paracoccidioidomicose, Micoses sistêmicas, Virulência, Gravidade, Modelo murino, Sequenciamento