Técnica de inmunoperoxidasa en miocardiopatia chagásica crônica experimental

Detalhes bibliográficos
Autor(a) principal: Maria Celina Morales, M.d.
Data de Publicação: 1987
Outros Autores: José Milei, M.D.
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Revista do Instituto de Medicina Tropical de São Paulo
Texto Completo: https://www.revistas.usp.br/rimtsp/article/view/28494
Resumo: 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.
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spelling Técnica de inmunoperoxidasa en miocardiopatia chagásica crônica experimental Chagas' diseaseImmunoperoxidase techniqueChronic myocarditis 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. 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. Universidade de São Paulo. Instituto de Medicina Tropical de São Paulo1987-04-01info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionapplication/pdfhttps://www.revistas.usp.br/rimtsp/article/view/28494Revista do Instituto de Medicina Tropical de São Paulo; Vol. 29 No. 2 (1987); 67-75 Revista do Instituto de Medicina Tropical de São Paulo; Vol. 29 Núm. 2 (1987); 67-75 Revista do Instituto de Medicina Tropical de São Paulo; v. 29 n. 2 (1987); 67-75 1678-99460036-4665reponame:Revista do Instituto de Medicina Tropical de São Pauloinstname:Instituto de Medicina Tropical (IMT)instacron:IMTenghttps://www.revistas.usp.br/rimtsp/article/view/28494/30347Copyright (c) 2018 Revista do Instituto de Medicina Tropical de São Pauloinfo:eu-repo/semantics/openAccessMaria Celina Morales, M.d. José Milei, M.D. 2012-07-02T00:56:56Zoai:revistas.usp.br:article/28494Revistahttp://www.revistas.usp.br/rimtsp/indexPUBhttps://www.revistas.usp.br/rimtsp/oai||revimtsp@usp.br1678-99460036-4665opendoar:2022-12-13T16:50:13.373933Revista do Instituto de Medicina Tropical de São Paulo - Instituto de Medicina Tropical (IMT)true
dc.title.none.fl_str_mv Técnica de inmunoperoxidasa en miocardiopatia chagásica crônica experimental
title Técnica de inmunoperoxidasa en miocardiopatia chagásica crônica experimental
spellingShingle Técnica de inmunoperoxidasa en miocardiopatia chagásica crônica experimental
Maria Celina Morales, M.d.
Chagas' disease
Immunoperoxidase technique
Chronic myocarditis
title_short Técnica de inmunoperoxidasa en miocardiopatia chagásica crônica experimental
title_full Técnica de inmunoperoxidasa en miocardiopatia chagásica crônica experimental
title_fullStr Técnica de inmunoperoxidasa en miocardiopatia chagásica crônica experimental
title_full_unstemmed Técnica de inmunoperoxidasa en miocardiopatia chagásica crônica experimental
title_sort Técnica de inmunoperoxidasa en miocardiopatia chagásica crônica experimental
author Maria Celina Morales, M.d.
author_facet Maria Celina Morales, M.d.
José Milei, M.D.
author_role author
author2 José Milei, M.D.
author2_role author
dc.contributor.author.fl_str_mv Maria Celina Morales, M.d.
José Milei, M.D.
dc.subject.por.fl_str_mv Chagas' disease
Immunoperoxidase technique
Chronic myocarditis
topic Chagas' disease
Immunoperoxidase technique
Chronic myocarditis
description 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.
publishDate 1987
dc.date.none.fl_str_mv 1987-04-01
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dc.identifier.uri.fl_str_mv https://www.revistas.usp.br/rimtsp/article/view/28494
url https://www.revistas.usp.br/rimtsp/article/view/28494
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv https://www.revistas.usp.br/rimtsp/article/view/28494/30347
dc.rights.driver.fl_str_mv Copyright (c) 2018 Revista do Instituto de Medicina Tropical de São Paulo
info:eu-repo/semantics/openAccess
rights_invalid_str_mv Copyright (c) 2018 Revista do Instituto de Medicina Tropical de São Paulo
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Universidade de São Paulo. Instituto de Medicina Tropical de São Paulo
publisher.none.fl_str_mv Universidade de São Paulo. Instituto de Medicina Tropical de São Paulo
dc.source.none.fl_str_mv Revista do Instituto de Medicina Tropical de São Paulo; Vol. 29 No. 2 (1987); 67-75
Revista do Instituto de Medicina Tropical de São Paulo; Vol. 29 Núm. 2 (1987); 67-75
Revista do Instituto de Medicina Tropical de São Paulo; v. 29 n. 2 (1987); 67-75
1678-9946
0036-4665
reponame:Revista do Instituto de Medicina Tropical de São Paulo
instname:Instituto de Medicina Tropical (IMT)
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