Revista do Instituto de Medicina Tropical de São Paulo, Volume: 29, Issue: 2, Published: 1987
  • Immunoperoxidase technique in experimental chronic chagasic myocarditis Original Articles

    Maria Celina Morales, M.d.; José Milei, M.D.

    Abstract in Spanish:

    La enfermedad de Chagas ha sido considerada como una de las causas más frecuentes de miocarditis crônica. Siendo descriptas Ias alteraciones inmunológicas, como patogenia para este tipo de enfermedad. Por tal motivo, se empleó la técnica de inmunoperoxidasa para la detección de depósitos de inmunoglobulinas en la miocardiopatía chagásica crônica experimental. Se utilizaron 41 ratones Swiss de 3 meses de vida, los mismos fueron inoculados intraperitonealmente con dosis entre 10 y 10(5); tripomastigotas de la cepa Tulahuen. La reinoculación se realizo 1 mes después con dosis entre 10² y 10(5); siendo sacrificados a los 6 (n=21) y 9 meses (n=9) de la primera inoculación, prévios estúdios elect roçar diográficos. Posteriormente se estudiaron los miocardios y músculos esqueléticos con técnicas histológicas de rutina, inmunoperoxidasa (método peroxidasa anti-peroxidasa) e inmunofluorescencia (directa e indirecta). Los métodos más sensibles para la detección de la enfermedad de Chagas crônica resultaron ser los estúdios histológicos (73%) y la electrocar-diografia (83%) a los 6 meses pi.y (89%) a los 9 meses p.i. (pos-infección). Se observaron diferentes alteraciones miocárdicas, desde infiltrados linfocitarios leves y focales en intersticio hasta reemplazo de miocitos por tejido conectivo. Los miocardio auriculares (21/23,91%) fueron más afectados que los ventrículos (9/23, 39%); mientras que las típicos quistes chagásicos resultaron excepcionales Los músculos esqueléticos (11/18 Y 7/9) presentaron distintos grados de lesión histológica, desdes leves a extensos infiltrados linfoplasmocitarios con presencia de fibras necróticas. Mientras que con la técnioa de inmunoperoxidasa, los antígenos se revelaron como depósitos granulares intracitoplasmáticos difusamente distribuidos, tanto para IgG como para Ig totales. La coincidência entre este método y Ias lesiones musculares histológicas fueron 11/18 (61%) a los 6 meses p.i.y 6/8 (75%) a los 9 meses p.i. Por otra parte, los depósitos de lg totales en corazón se observaron dispuestos difusamente, en forma finamente granular dentro de los miocitos ventriculares La coincidência entre ambas técnicas (inmunoperoxidasa e histología) resultó ser dei 36% y 66% para los animales sacrificados a los 6 y 9 meses p.i. respectivamente. Este fenômeno inmunológico se incremento notablemente con el curso crônico de la enfermedad. Con respecto a la sensibilidad, tanto la inmunoperoxidasa como la inmunofluorescencia directa fueron altamente sensibles en músculo esquelético (100%, p 0,01). Por otra parte, la técnica de inmunofluorescencia directa en corazón, evidencio pobres resultados, mientras que el método peroxidasa anti-peroxidasa incrementó su sensibilidad de 21,4% a los 6 meses pi. al 66.6% a los 9 meses p.i. (p 0.001). Este modelo experimental, en el cual se observan reacciones inmunológicas reveladas por la técnica de inmunoperoxidasa, podría resultar de utilidad, considerando la necesidad de obtener una vacuna adecuada para prevenir la miocardiopatía chagásica.

    Abstract in English:

    Chagas'disease has been described as the commonest form of chronic myocarditis. An immunologic pathogenesis has been discribed for this form of the disease. So far, no immunoperoxidase technique has been used for the detection of immunological deposits in chronic experimental Chagas'myocardiopathy. Forty-one Swiss mice, three months old were inoculated intraperitoneally with doses between 10 and 10(5) Tulahuen trypomastigotes. Mice were reinoculated one month after with doses between 10² and 10(5) and sacrificed at 6 (n=21) and 9 months (n=9) after the first inoculation. ECGs were recorded before sacrifice. Immunoperoxidase technique (peroxidase-antiperoxidase method), immunofluorescence (direct and indirect) as well as histological studies were performed in myocardiums and skeletal muscles of the surviving animals. The most sensitive methods for detecting chronic chagasic infection were the routine histologic studies (73%) and the ECGs 83% and 89% on 6 and 9 mo. post-infected mice, respectively. Myocardial involvement varied from interstitial mild focal lymphocyte infiltrates up to replacement of myocytes by loose connective tissue. Atrial myocardiums (21/23, 91%) were more affected than ventricles (9/23, 39%). Typical chagasic nests were rarely found. Skeletal muscle involvement (11/18 and 7/9) varied from mild to extensive lymphocyte and plasmacell infiltrates, and necrotic fibers. The involved antigen were shown in skeletal muscles by the immunoperoxidase technique as diffusely arranged granular intracytoplasmatic deposit for both IgC and total immunoglobulins. The coincidence between this technique and histologic muscle lesions was 11/18 (61(%) in 6 mo. and 6/8 (75%) at 9 mo. post-infection. In heart, delicate granular deposits of total immunoglobulins were seen diffusely arranged within the ventricular myocytes; coincidence between immunoperoxidase technique anl histologic involvement increased from 36 to 66% in animals sacrifeced 6 and 9 mo. post-infection. This strongly stressed the increase of immunologic phenomena with the chronification of infection. Concerning sensitivity, immunoperoxidase and direct immunofluorescence were highly sensitive in skeletal muscle (100%, p < 0.01). Conversely, direct immunofluorescence technique showed poor results in heart while immunoperoxidase increased its sensitivity from 21.4% (at 6 mo.) to 66.6% (at 9 mo.) post-infection (p < 0.001). Considering the necessity of obtaining an adequate vaccine in order to prevent this disease an experimental model like this, rendering immunological reactions as revealed by the immunoperoxidase technique, would be useful.
  • Periphyton used as food for Biomphalaria tenagophila (Gastropoda, Planorbidae) Original Articles

    Santos, M. B. L.; Freitas, J. R.

    Abstract in Portuguese:

    Moluscos criados em gaiolas colonizadas por perifíton durante vinte dias cresceram em diâmetro de concha de 5,0 mm a 8,6 mm, apresentaram uma sobrevivência de 75% e uma média de 0,5 desova molusco/dia.

    Abstract in English:

    Snails reared in cages colonized by periphyton grew from 5.0mm to 8.8mm shell diameter, each laid 0.5 batch of eggs per day and the overall survival during the period was 75%
  • Trypanosoma cruzi: expression of antigenic component 5 among 35 laboratory clones obtained from 18 different isozymic variants Original Articles

    Breniere, Simone F.; Revollo, Susana; Caillard, Thierry; Velatte, Eric; Legrand, Dominique; Afchain, Daniel; Desjeux, Philippe

    Abstract in Portuguese:

    Dois anticorpos monoclonais anticomponente 5 de Trypanosoma cruzi (I-35/115 e II-190/30) foram testados respectivamente em IFA e ELISA sobre 35 clones de T. cruzi isolados no laboratório. Entre estes 35 clones testados, 18 perfis isoenzímicos diferentes puderam ser detectados. Todos os clones foram reconhecidos exceto um clone que não reagiu com o anticor po monoclonal II-190/30. Estes resultados são a favor da expressão constante do componente 5 no seio do taxón T. cruzi.

    Abstract in English:

    Two monoclonal antibodies anti-component 5 of Trypanosoma cruzi (I-35/115 and II-190/30) were tested in IFA and ELISA respectively against 35 T. cruzi laboratory clones. Among the 35 clones tested, 18 different isozyme patterns were detected. All clones were recognized by both monoclonal antibodies except one clone which did not react with II-190/30. These results support the universal expression of specific component 5 within the taxon T. cruzi.
  • Molluscum contagiosum: serology and electron microscopy findings in twenty one patients Original Articles

    Fonseca, M. E. F.; Machado, R. D.; Liberto, M. I. M.; Marcolino, G.

    Abstract in Portuguese:

    Vinte e um casos de molusco contagioso foram escolhidos para serem analisados por testes sorológicos e de microscopia eletrônica. O vírus do molusco foi detectado nas crostas e observados também nos vacúolos formados dentro dos queratinócitos. Os pacientes desenvolveram anticorpos fixadores do complemento para o vírus.

    Abstract in English:

    Twenty one cases of molluscum contagiosum virus disease were collected for electron microscopical and serological tests. Molluscum virus was detected in the crust, inside the vacuoles formed in the keratinocytes cells. The patients developed specific antibodies to the virus detected by complement fixation test.
  • Histopathology of cutaneous and mucosal lesions in human paracoccidioidomycosis Original Articles

    Uribe, Fabio; Zuluaga, Angela I.; León, Walter; Restrepo, Angela

    Abstract in Portuguese:

    Biopsias de lesões cutânea e mucosa de 40 pacientes com paracoccidioidomicose ativa foram estudadas em seu aspecto histopatológico. Todos os casos mostravam inflamação granulomatosa crônica e 38 também apresentavam supuração; este quadro correspondia ao granuloma micótico misto (MMG). A eliminação do parasita ocorre através da formação de edema progressivo, acompanhado por exocitose. O edema evolui a espongiose, microvesículas e microabscesses que não só contém o fungo, mas também vários elementos celulares. As células encarregadas do processo fagocitário eram essencialmente células gigantes de Langhams; polimorfonucleares, células epitelióides e células gigantes de corpos estranhos eram fagócitos pobres. Um achado adicional foi a presença de fibrose na maioria das biópsias.

    Abstract in English:

    Biopsies from cutaneous and mucosal lesions from 40 patients with active paracoccidioidomycosis, were studied histopathologically. All cases exhibited chronic granulomatous inflammation and 38 also presented suppuration; this picture corresponded to the mixed mycotic granuloma (MMG). Pseudoepitheliomatous hyperplasia and the transepidermic (or epithelial) elimination of the parasite, were observed in all cases. In paracoccidioidomycosis elimination takes place through formation of progressive edema, accompained by exocytosis. The edema gives rise to spongiosis, microvesicles and microabscesses which not only contain the fungus but also, various cellular elements. Cells in charge of the phagocytic process were essentialy Langhans giant cells; PMN's, epithelioid and foreign body giant cells were poor phagocytes. An additional finding was the presence of fibrosis in most biopsies.
  • Immunoglobulins and C3 in the P. brasiliensis granuloma Original Articles

    Biagioni, Lilian M. V.; Orsi, Sonia; Chamma, Luiz G.; Sadatsune, Terue; Frango, Marcello

    Abstract in Portuguese:

    Utilizou-se o modelo experimental de paracoccidioidomicose, em camundongos, induzida pela inoculação endovenosa de suspensão de formas cerebriformes do P. brasiliensis (cepa Bt2; 1x10(6) formas viáveis/animal), para avaliar, após 2, 4, 8, 16 e 20 semanas: 1. A presença de imunoglobulinas e C3 nos granulomas pulmonares, por imunofluorescência direta; 2. A resposta imune humoral (imunodifusão) e celular (teste do coxim plantar), e 3. A histopatologia das lesões. Os camundongos apresentaram resposta imunocelular positiva desde a 2a. semana, com depressão transitória na 16a. semana, e anticorpos desde a 4a. semana, com pico na 16a. semana. Os granulomas pulmonares foram epitelióides, com numerosos fungos e microabscesses; a extensão das lesões foi progressiva até a 16a. semana, com regressão discreta na 20a. semana. Desde a 2a. semana, houve deposição de IgG e C3 na parede dos fungos no interior dos granulomas e a presença de células IgG positivas no halo linfomononuclear periférico; estes achados foram máximos entre a 4a. e 16a. semanas. Não se detectou depósito de IgG e C3 no interstício dos granulomas. IgG e C3 parecem exercer papel precoce e importante na defesa do hospedeiro contra o P. brasiliensis, contribuindo possivelmente para a destruição dos fungos e bloqueando a difusão de antígenos para fora dos granulomas.

    Abstract in English:

    The experimental model of paracoccidioidomycosis induced in mice by the intravenous injection of yeast-forms of P. brasiliensis (Bt2 strain; 1 x 10(6) viable fungi/animal) was used to evaluate sequentially 2, 4, 8, 16 and 20 weeks after inoculation: 1. The presence of immunoglobulins and C3 in the pulmonary granuloma-ta, by direct immunofluorescence; 2. The humoral (immunodiffusion test) and the cellular (footpad sweeling test) immune response; 3. The histopathology of lesions. The cell-immune response was positive since week 2, showing a transitory depression at week 16. Specific antibodies were first detected at week 4 and peaked at week 16. At histology, epithelioid granulomas with numerous fungi and polymorphonuclear agreggates were seen. The lungs showed progressive involvement up to week 16, with little decrease at week 20. From week 2 on, there were deposits of IgG and C3 around fungal walls within the granulomas and IgG stained cells among the mononuclear cell peripheral halo. Interstitital immunoglobulins and C3 deposits in the granulomas were not letected. IgG and C3 seen to play an early an important role in. the host defenses against P. brasiliensis by possibly cooperating in the killing of parasites and blocking the antigenic diffusion.
  • Cell mediated immune response in human antirabies revaccination Original Articles

    Veiga, Débora Regina; Zanetti, Carlos Roberto; Mendes, Nelson Figueiredo; Pereira, Octávio Augusto de Carvalho

    Abstract in Portuguese:

    Foi estudada a ocorrência de resposta imune celular na re imunização anti-rábica humana. Usou-se a vacina Fuenzalida & Palácios a qual é rotineiramente usada no Brasil. A resposta imune celular foi avaliada pelo índice de transformação blástica obtido em cultura de sangue total, na presença de antígeno rábico e antígeno controle (sistema nervoso) . O grupo em estudo constitui se de 11 voluntários submetido a re vacinação, enquanto três outros, submetidos a primo-vacinação foram utilizados como grupo controle. Detectou-se uma clara resposta imune celular secundária contra antígeno rábico em todos os voluntários revacinados, cujas respostas foram mais rápidas e mais intensas do que as do grupo controle. Observou-se também, resposta ao antígeno controle em todos os indivíduos do grupo de revacinados. No grupo de primovacinados, resposta ao antígeno controle foi fracamente observada em somente um indivíduo.

    Abstract in English:

    The occurrence of secondary cell mediated immune response (CMI) in human antirabies immunization was studied. The Puenzalida & Palácios vaccine was used because it is routinely used in Brazil. CMI was evaluated by lymphoblastic transformation indices obtained in whole blood culture in the presence of rabies and control (nervous tissue) antigens. Eleven volunteers submitted to revaccination constituted the group under study, while three other volunteers submitted primo vaccination were utilized as control group. A clear secondary CMI to rabies antigen was detected in all the revaccinated volunteers who showed earlier and more intense response than the control group. Response to the control antigen, however, present in all the components of the first group was not detectable in two out of the three primovaccinated and very low in the third one.
  • The presence of antibodies for hepatitis a virus in amazonia Didelphis marsupialis (Vertebrata, Marsupialia) Original Articles

    Soares, Manoel do Carmo P.; Bensabath, Gilberta; Rosa, Amélia P. A. Travassos da

    Abstract in Portuguese:

    Foi detectada a presença de anticorpos para o vírus da Hepatite A (anti-VHA), utilizando-se a técnica de enzimaimunoensaio, no soro de 6 (18,75%) entre 32 Didelphis marsupialis capturados na Região Amazônica. Não foi detectado anti-VHA em soros de outros 136 animais silvestres incluindo pequenos roedores, répteis e outros marsupiais.

    Abstract in English:

    Anti-HAV was detected by enzyme - immunoassay in sera collected from 6 (18,75%) of 32 Didelphis marsupialis trapped in the Amazon region. No anti-HAV were found in the sera from 136 other wild animals, including small rodents, reptiles and other marsupials.
  • Human disease in ribeira valley, brazil caused by caraparu, a group c arbovirus - Report of a case Case Report

    Iversson, Lygia Busch; Rosa, Amélia P. A. Travassos da; Coimbra, Terezinha Lisieux M.; Ferreira, Ivani Bisordi; Nassar, Elza da Silva

    Abstract in Portuguese:

    Relatam-se os dados clínicos e laboratoriais de um caso de moléstia humana causada por vírus idêntico ou antigenicamente muito relacionado ao arbovirus do Grupo C Caraparu, em um morador da região do Vale do Ribeira, Estado de São Paulo, sudeste do Brasil. O fato apresenta interesse médico sanitário pois embora existam evidências da presença de inúmeros arbovirus na área, os únicos casos comprovados de doença por esses agentes foram os de encefalite pelo vírus Rocio durante a epidemia ocorrida em 1975-1977. Os resultados dos testes sorológicos sugerem diferença antigênica entre as cepas de vírus Caraparu isoladas nos Estados de São Paulo e Pará e proximidade antigênica entre a cepa de Caraparu de São Paulo e o vírus Bruconha.

    Abstract in English:

    The clinical and laboratory data of a disease in a resident of Ribeira Valley, São Paulo State, southeastern Brazil, caused by an agent close or identical to Caraparu, a Group C arbovirus, was described. Although there is evidence of an intensive circulation of several arboviruses in the area, no diagnosis of human disease by these agents has been made, except the encephalitis cases caused by Rocio virus during an epidemic in 1975-1977. An antigenic difference between Caraparu strains isolated in São Paulo and in Pará States and a close antigenic relationship between Caraparu strain from São Paulo and Bruconha virus were suggested by the serological tests.
  • Untitled document Case Report

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