Vinte e sete pacientes portadores de paracoccidioidomicose (PCM) foram tratados com itraconazole (100-200 mg/dia no primeiro mês e 100 mg/dia até 6-8 meses) e avaliados sob o ponto de vista clínico e sorológico, até 3 e meio anos após o início do tratamento, utilizando-se os testes de Dot-blot e ELISA para medir os títulos de anticorpos IgG, IgA e IgM anti-P. brasiliensis, e Western-blot para determinar os anticorpos IgG, IgA e IgM contra os componentes antigênicos do fungo. Antes do tratamento, 81,5% (Dot-blot) e 84% (ELISA) dos pacientes apresentaram títulos elevados de anticorpos IgG anti-P. brasiliensis, que decresceram levemente com o tratamento. Por outro lado, as porcentagens de soros pré-tratamento com títulos elevados para anticorpos IgA e IgM foram menores (51,9% e 51,8%: Dot-blot; 16% e 36 %: ELISA, respectivamente); com o tratamento, entretanto, estes títulos tenderam mais frequentemente a se negativar. Antes do tratamento, as porcentagens de positividade para anticorpos IgG, IgA e IgM, avaliados por Western-blot, foram 96%, 20,8% e 41,6%, respectivamente. Componentes antigênicos de massas moleculares variando entre 16-78 kDa, 21-76 kDa e 27-78 kDa reagiram com anticorpos das classes IgG, IgA e IgM, respectivamente, As frações antigênicas com massas moleculares de 27, 33 e 43 kDa foram as mais frequentemente reativas com anticorpos da classe IgG, e a de 70 kDa para anticorpos IgA e IgM. Todos os pacientes apresentaram remissão da sintomatologia com o tratamento, durante o período de estudo. Os dados do presente trabalho confirmam a diversidade e a complexidade da resposta humoral dos pacientes com PCM e reforçam a importância de se utilizar diferentes testes sorológicos para se detectar anticorpos IgG, IgA e IgM anti- P. brasiliensis.

Twenty-seven mycologically proven cases of paracoccidioidomycosis (PCM) were treated with itraconazole (100-200 mg/day in month 1 and 100 mg/day until month 6-8) and evaluated clinically and serologically, up to 3.5 years post-therapy, using Dot-blot and ELISA for measuring the titers of IgG, IgA and IgM anti-P. brasiliensis antibodies and Western-blot for determining IgG, IgA and IgM antibodies against the antigen components of the fungus. Before treatment, 81.5% (Dot-blot) and 84% (ELISA) of the patients presented elevated IgG anti-P. brasiliensis antibody titers which dropped slightly with treatment. On the other hand, the percentages of pre-treatment high-titered sera for IgA and IgM anti-P.brasiliensis were lower (5l.9% and 5l.8%: Dot-blot; 16.5 and 36%: ELISA, respectively) but the titers tended to become negative more frequently with treatment. Prior to treatment, the percentages of positivity for IgG, IgA and IgM anti-P.brasiliensis antibodies in Western-blot were 96%, 20.8% and 41.6%, respectively. Antigens with molecular weights varying from 16-78 kDa, from 21-76 kDa and from 27-78 kDa were reactive for IgG, IgA and IgM antibodies, respectively. The most frequently reactive antigenic components had molecular weights of 27, 33 and 43 kDa for IgG, and 70 for IgA and IgM antibodies. During the period of study, the patients responded well to treatment. The present data confirm the diversity and complexity of the humoral response in PCM, and the importance of utilizing different serological tests to detect IgG, IgA and IgM anti-P. brasiliensis antibodies

The objectives of the present study were to optimize the protocol of mouse immunization with Paracoccidioides brasiliensis antigens (Rifkind's protocol) and to test the modulation effect of cyclophosphamide (Cy) on the delayed hypersensitivity response (DHR) of immunized animals. Experiments were carried out using one to four immunizing doses of either crude particulate P. brasiliensis antigen or yeast-cell antigen, followed by DHR test four or seven days after the last immunizing dose. The data demonstrated that an immunizing dose already elicited response; higher DHR indices were obtained with two or three immunizing doses; there were no differences between DHR indices of animals challenged four or seven days after the last dose. Overall the inoculation of two or three doses of the yeast-cell antigen, which is easier to prepare, and DHR test at day 4 simplify the original Rifkind's immunization protocol and shorten the duration of the experiments. The modulation effect of Cy on DHR was assayed with administration of 2.5, 20 and 100 mg/kg weight at seven day intervals starting from day 4 prior to the first immunizing dose. Only the treatment with 2.5 mg Cy increased the DHR indices. Treatment with 100 mg Cy inhibited the DHR, whereas 20 mg Cy did not affect the DHR indices. Results suggest an immunostimulating effect of low dose of Cy on the DHR of mice immunized with P. brasiliensis antigens.

Com o objetivo de padronizar modelo experimental de imunossupressão foram selecionados dois lotes de 50 camundongos impúberes, cada um dividido em grupos de 10, e submetidos a ação de dexametasona. O estudo foi dividido em dois experimentos. No primeiro experimento foi utilizado o primeiro lote de animais e foram estabelecidas as condições para obtenção de modelo de imunossupressão em camundongos mediante administração de dexametasona diluída na água de consumo e ingerida ad libitum pelos animais. Entre os critérios utilizados para avaliar a vigência de imunossupressão são realçados: a ocorrência de eriçamento de pelos, leucopenia por diminuição do número de linfócitos e hipotrofia do baço comparativamente aos controles, com hipoplasia intensa de todas as estruturas linfóides. No segundo experimento foi empregado o segundo lote de animais e foi estudado o comportamento do modelo experimental estabelecido. Os animais receberam, por via intraperitonial, número aproximado delO6 leveduras com os caracteres morfológicos do Cryptococcus neoformans, provenientes de culturas de cepas patogênicas ao homem e isoladas do líquido cefalorraqueano de pacientes com neurocriptococose. Apesar de haverem sido identificadas leveduras em cortes histológicos de pulmão, fígado e baço em todos os camundongos inoculados, somente ocorreu o óbito naqueles em que havia acometimento do sistema nervoso central pela levedura (aproximadamente 50% dos animais). A discussão dos resultados obtidos e seu significado inclui considerações sobre o possível papel desempenhado pela função barreira hêmato-encefálica no evento constatado.

In order to accomplish a model for immunossupression in experimental conditions two lots of 50 young mice were divided into groups of 10, and submitted to dexamethasone ingestion. Two experiments were considered in the study. In the first experiment conditions for immunossupression were established in 50 mice, by per os supply of several concentrations of dexamethasone in the water given for normal daily intake ad libitum. Criteria of immunossupression considered include: hair standing on end; leukopenia by decrease of lymphocyte population; spleen atrophy in relation to controls, with severe hypoplasy of all lymphoid structures. In the second experiment another lot of 50 mice in similar conditions established for the first experiment and concerning the immunossupression were submitted to intraperitonial innoculation of near 10e Cryptococcus neoformans. Culture suspensions of cryptococci isolated from cerebrospinal fluid samples of human neurocryptococcosis cases were used for this purpose. Although fungi could be recognized in lungs, liver and spleen from every mice innoculated, only those with central nervous system involvement (near by an half of all) died. The discussion includes considerations about the role of the blood-brain barrier function in the findings registered.