Trypanosoma cruzi: circulating antigens

Circulating antigens were detected in sera of mice experimentally infected with a high close of Trypanosoma cruzi by reaction with sera from chronically infected mice. The immunodiffusion reaction between homologous acute and chronic sera produced four precipitation lines. By reaction with chronic mouse serum, circulating antingens were detected in sera from heavily infected hamsters, dogs, rabbits and in sera from chagasic patients. A reaction was also found in urine from acutely infected mice and dogs. Trypanosoma cruzi exoantigen was detected in trypanosome culture medium and in the supernatant of infected cell cultures. Attempts to isolate the antigens are described.

Antigenos "particulares" a cepas do Trypanosoma cruzi: demonstracao por imunoeletroforese bidimensional

Os extratos solúveis das cepas Y, São Felipe e Colombiana do Trypanosoma cruzi foram analisados contra seus antissoros homólogos e heterólogos por imunoeletroforese bidimensional e imunoeletroforese bidimensional com gel intermediário. Os resultados revelaram a existência de pelo menos 35 linhas de precipitação na cepa Y (32 de migração anódica e três de migração catódica), 24 linhas de precipitação anódica na cepa São Felipe e 22 na cepa Colombiana. Estas duas últimas cepas não apresentaram antígenos de migração catódica. Estes antígenos de migração catódica foram considerados "particulares" a cepa Y uma vez que quando testados contra antissoros heterólogos não observamos linhas de precipitação. Através do uso da imunoeletroforese bidimensional com gel intermediário foram evidenciados cinco antígenos particulares à cepa Y quando comparada à São Felipe e oito quando comparada à cepa Colombiana. A cepa São Felipe mostrou um único antígeno particular quando comparada à cepa Colombiana porém, não se evidenciou nenhum antígeno particular quando a cepa São Felipe foi analisada contra o antissoro da cepa Y. A cepa Colombiana não demonstrou nenhum antígeno particular nem quando comparada à cepa Y nem à São Felipe. Nossos resultados revelam que cepas pertencentes a diferentes tipos quanto ao comportamento morfobiológico e histopatológico também demonstram uma acentuada diferença quanto a seus componentes antigênicos.

Ensayos metodologicos para la investigacion de reservorios de Leishmania spp en los Andes venezolanos

Se describen dos técnicas, presuntiva y confirmativa, para la investigación de mamíferos que pudieran ser reservorios de Leishmania que parasitan al hombre. Se investigan los cambios en los títulos de inmovilización y aglutinación de promastigotos de cultivo por los sueros de animales normales y expuestos una o varias veces a la inoculación intradérmica de pequeñas dosis de promastigotos vivos. Se registra una caída de los títulos de aglutinación en los sueros de hamsteres, de Holochilus venezuelae y de Didelphis marsupialis después de la inoculación con L. mexicana mexicana de Panamá y de L. gamhami de la región de los Andes venezolanos. Se discute la natureza de estos fenómenos. Se han hecho xenodiagnósticos con Lutzomyia townsendi en Holochilus venezuelae y Sigmodon hispidus infectados experimentalmente com L. mexicana mexicana, L. mexicana amazonensis, L. braziliensis y L. garnhami. Las pruebas fueron leidas mediante el examen microscópico de las gotitas de heces excretadas entre las 108 y 132 horas después de la ingesta infectante, tras colorearlas con Giemsa. Se obtuvieron resultados positivos en 23% de los experimentos usando mamíferos con lesiones localizadas, dejando a los flebótomos ingurgitarse libremente sobre animales anestesiados que poseian una hasta varias lesiones localizadas.

Produção e padronização dos antígenos de Paracoccidioides brasiliensis (Pb), Histoplasma capsulatum (Hc) e Aspergillus fumigatus (Af) para uso no imunodiagnóstico: comparação entre as técnicas de imunodifusão e imunoeletroosmoforese

Foram produzidos antígenos solúveis de P. brasiliensis, H. capsulatum e A. fumigatus e padronizados nas técnicas de imunodifusão dupla (IDD) e imunoeletroosmoforese (IEOF). A especificidade dos antígenos foi testada utilizando-se 96 soros de pacientes com paracoccidioidomicose, histoplasmose, aspergilose, candidíase sistêmica, esporotricose, tuberculose, neoplasia pulmonar, leishmaniose tegumentar e visceral e em 18 indivíduos sadios. Na IDD, a especificidade foi de 100% usando-se como critério de positividade linhas de precipitação com identidade total com soro de referência. Entretanto na IEOF, a especificidade variou de acordo com o antígeno testado, sendo observadas reações cruzadas com antígeno de P. brasiliensis frente a soros de pacientes com histoplasmose (16,7%) e leishmaniose tegumentar (10%) e com antígeno de H. capsulatum frente a soros de pacientes com paracoccidioidomicose (31,8%) e leishmaniose tegumentar (10%). Ambas as técnicas mostraram a mesma sensibilidade para o sorodiagnóstico de paracoccidioidomicose, histoplasmose e aspergilose, respectivamente 100%, 83,3% e 100%. A grande sensibilidade e especificidade da IDD observadas nos soros desses pacientes, aliadas à fácil reprodutibilidade e baixo custo, fazem esta técnica muito apropriada como procedimento de rotina, para a triagem de pacientes sintomáticos respiratórios.

Patterns of serologic response to human immunodeficiency virus type 1 (HIV-1) in brazilians with different clinical forms of HIV infection

In order to investigate the IgG HIV-1 antibodies rectivity to structural components of the virus, 85 sera from infected Brazilians, comprising the total spectrum of HIV infection, were analysed by Western blot assay. The sera were confirmed as being positive to HIV with enzyme linked immuno assay (ELISA) and indirect immunofluorescence (IIF). Although the sera from patients reacted less intensively to the gag polypeptide of 55KDa, no distinctive antigen reaction patterns were observed between sera patients with different clinical forms. Because of the higher frequency of reactivity to the gag p24 in AIDS patients, the patterns of anti-HIV IgG responses are similar to those observed in their African counterparts.

Experimental chagas' disease in rhesus monkeys. I. Clinical parasitological, hematological and anatomo-pathological studies in the acute and indeterminate phase of the disease

Rhesus monkeys (macaca mulatta) were infected subcutaneously with 1.0 x 10**4 to 1.5 x 10**4 metacyclic trypomastigotes of Trypanosoma cruzi (Colombian strain). Parasitological and immunological parameters were evaluated in these animals for periods of 1 month to over 3 years. a chagona was observed between the 3 rd and the 13th day after infection (a.i) and patent parasitaemia between the 13th and 59th day a.i.. Thereafter, parasites were demonstrated only by haemoculture and/or xenodiagnosis. Circulating specifc IgM and IgC antibodies were observed as early as in the 2nd week a. i. IgG levels persisted until the end of the expriment, but IgM antibodies were detectable nine months a. i. Haematological alterations comprised leucocytosis and lymphocytosis. Eletrocardiographic alterations were minor and transient, similar to those observe in non-lethal human acute Chagas' myocarditis. Myocarditis and myositis, characterized by multiple foci of lympho-histiocyte inflammatory infiltrate, were present in monkeys sacrificed on the 41 th, 70th and 76 th day but not in the animal sacrificed 3 years and 3 months a. i.. The results suggest that Chagas' disease in rhesus monkeys reproduces the acute and indeterminate phases of human Chagas' disease.

Trypanosoma cruzi: experimental Chagas' disease in Rhesus monkeys. II. Ultraestructural and cytochemical studies of peroxidase and acid phosphatase activities

Ultrastructural and cytochemical studies of peroxidase and acid phosphatase were performed in skin, lymph node and heart muscle tissue of thesus monkeys with experimental Chagas's disease. At the site of inoculation ther was a proliferative reaction with the presence of immature macrophages revealed by peroxidase technique. At the lymph node a difuse inflammatory exudate with mononuclear cells, fibroblasts and immature activated macrophages reproduces the human patrtern of acute Chagas' disease inflamatory lesions. The hearth muscle cells present different degrees of degenerative alterations and a striking increase in the number of lysosomal profiles that exhibit acid hydrolase reaction product. A strong inflammatory reaction was present due to lymphocytic infiltrate or due to eosinophil granulocytes associated to ruptured cells. The present study provides some experimental evidences that the monkey model could be used as a reliable model to characterize histopathological alterations of the human disease.

Molecular and biological diversity of HIV-1 in Brazil

To determine the genomic polymorphism and biological properties present in HIV-1 Brazilian isolates, were analyzed five viral isolates obtained from patients residing in Rio de Janeiro (P1 and P5), São Paulo (P3) and Bahia (P2 and P4) states. For each viral isolate in vitro characteristics such as replication rate, syncytium-inducing capacity and cell death were observed in lymphoblastoid (H9, CEM and peripheral blood mononuclear cells) as well as monocytoid (U937) cells. In addition, the evaluation of the restriction fragment lenght polymorphism of these isolates was also performed using a panel of endonucleases such as Hind III, Bgl II, Sac I, Pst I, Kpn I and Eco RI. One of the isolates (P1), showed the highest phenotypic and genotypic divergence, when compared to others. The results found suggest a HIV heterogeneity in Brazil similar to that already described in other regions of the world.