Técnica alternativa para fechamento de placas de microtécnica em exames de histocompatibilidade HLA
Autor(a) principal: | |
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Data de Publicação: | 2022 |
Outros Autores: | , , |
Tipo de documento: | Artigo |
Idioma: | por |
Título da fonte: | Revista Brasileira de Cancerologia (Online) |
Texto Completo: | https://rbc.inca.gov.br/index.php/revista/article/view/2958 |
Resumo: | The HLA system consists in a group of cell membrane proteins: the HLA-A, B, C, DR, DQ and DP molecules. Arnong the main uses of this system we can find the farnily and random studies for bone marrow trans plantation, as well as the cross-matches before kidney transpiantations. An irnportant rnethodological advance in the study of the HLA system arised very soon, with the development of a rnicrolyrnphocytotoxiciiy technique, used until nowadays, in which the mononuclear blood cells bearing the HLA molecules are used as target for cytotoxicity reactions developed in specialized rnicrotrays. The final step of the serological rnethod consists to cover the tray with a glass cover slip and to read the reactions at an inverted light microscope. We present an alternative technique to dose the tray, which perrníts the utilization of a common light microscope. In this proposed method, a gel (Kaiser's rnodífied gelatin) is placed gently over the tray at the end of the usual technique, following eosin staining and formaline fixation. After that, the tray is placed, in inverted position, at a common light microscope and read in the same way as the usual rnethod. Twenty preliminary tests were perforrned unti/ now, with a very good reprodutibilily in terrns of reaction patterns; no mor phological altera tions were seen in these tests, when cornpared with the conventional technique. The use of this gelatin proved to be econornical, easy to perforrn and very reproducible. The trays covered with the gelatin were read again each two weeks for six rnonths, and we did not find any difference in the reaction patterns as well as in the cell morphology. |
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Técnica alternativa para fechamento de placas de microtécnica em exames de histocompatibilidade HLASistema HLASorologia HLAMicrolinfocitotoxicidadeTipagem HLASystemHLA SerologyMicroiymphocytotoxicityHLA TypingThe HLA system consists in a group of cell membrane proteins: the HLA-A, B, C, DR, DQ and DP molecules. Arnong the main uses of this system we can find the farnily and random studies for bone marrow trans plantation, as well as the cross-matches before kidney transpiantations. An irnportant rnethodological advance in the study of the HLA system arised very soon, with the development of a rnicrolyrnphocytotoxiciiy technique, used until nowadays, in which the mononuclear blood cells bearing the HLA molecules are used as target for cytotoxicity reactions developed in specialized rnicrotrays. The final step of the serological rnethod consists to cover the tray with a glass cover slip and to read the reactions at an inverted light microscope. We present an alternative technique to dose the tray, which perrníts the utilization of a common light microscope. In this proposed method, a gel (Kaiser's rnodífied gelatin) is placed gently over the tray at the end of the usual technique, following eosin staining and formaline fixation. After that, the tray is placed, in inverted position, at a common light microscope and read in the same way as the usual rnethod. Twenty preliminary tests were perforrned unti/ now, with a very good reprodutibilily in terrns of reaction patterns; no mor phological altera tions were seen in these tests, when cornpared with the conventional technique. The use of this gelatin proved to be econornical, easy to perforrn and very reproducible. The trays covered with the gelatin were read again each two weeks for six rnonths, and we did not find any difference in the reaction patterns as well as in the cell morphology.O Sistema HLA é composto por glicoproteínas integrais de membrana: as moléculas HLA A, B, C, DR, DO e DP. Entre as principais aplicações desse sistema encontram-se os estudos para os transplantes de medula óssea. Um importante avanço metodológico no estudo do sistema HLA surgiu com a descrição de um teste de microlinfocitotoxicidade: uma microtécnica empregada até os dias de hoje, na qual células portadoras dos HLA são utilizadas como alvo em reações de citotoxicidade desenvolvidas em microplacas específicas (placas Terasaki). A etapa final do método consiste na cobertura da placa com lamínula de vidro e leitura ao microscópio óptico de luz invertida. O presente trabalho apresenta uma técnica alternativa de fechamento da placa Terasaki que permite a leitura em microscópio óptico comum. Resume-se na colocação de um preparado tipo "gel" (Gelatina de Kaiser modificada) sobre a placa Terasaki ao término da metodologia usual, permitindo a inversão da mesma para leitura em microscópio óptico comum. O procedimento técnico de sorologia HLA é desenvolvido normalmente, e após coloração com eosina e fixação com formol neutro, a gelatina é dispensada na placa vagarosamente, devendo cobri-la homogeneamente. A preparação é deixada então à temperatura ambiente até que atinja o estado "gel". A leitura é feita em microscópio óptico comum, colocando-se a placa tampada com o fundo voltado para cima. Foram realizados 20 testes preliminares utilizando-se em paralelo a técnica convencional de fechamento e a técnica alternativa. A técnica mostrou-se econômica e de fácil execução, e a reprodutibilidade das leituras quando comparamos os dois métodos mostrou-se satisfatória. Nas placas em que se utilizou a Gelatina, novas leituras foram realizadas quinzenalmente durante 6 meses, e não se verificaram quaisquer alterações na morfologia celular nem no padrão de leitura.INCA2022-10-04info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionapplication/pdfhttps://rbc.inca.gov.br/index.php/revista/article/view/295810.32635/2176-9745.RBC.1995v41n4.2958Revista Brasileira de Cancerologia; Vol. 41 No. 4 (1995): Oct./Nov./Dec.; 271-274Revista Brasileira de Cancerologia; Vol. 41 Núm. 4 (1995): oct./nov./dic.; 271-274Revista Brasileira de Cancerologia; v. 41 n. 4 (1995): out./nov./dez.; 271-2742176-9745reponame:Revista Brasileira de Cancerologia (Online)instname:Instituto Nacional de Câncer José Alencar Gomes da Silva (INCA)instacron:INCAporhttps://rbc.inca.gov.br/index.php/revista/article/view/2958/1831https://creativecommons.org/licenses/by/4.0info:eu-repo/semantics/openAccessPaiva, Sérgio Ré dePaiva, Tádia Maria Cordeiro dePontes, Luciane Faria de SouzaMorínigo, Fábio Cupertino2023-06-19T13:19:31Zoai:rbc.inca.gov.br:article/2958Revistahttps://rbc.inca.gov.br/index.php/revistaPUBhttps://rbc.inca.gov.br/index.php/revista/oairbc@inca.gov.br0034-71162176-9745opendoar:2023-06-19T13:19:31Revista Brasileira de Cancerologia (Online) - Instituto Nacional de Câncer José Alencar Gomes da Silva (INCA)false |
dc.title.none.fl_str_mv |
Técnica alternativa para fechamento de placas de microtécnica em exames de histocompatibilidade HLA |
title |
Técnica alternativa para fechamento de placas de microtécnica em exames de histocompatibilidade HLA |
spellingShingle |
Técnica alternativa para fechamento de placas de microtécnica em exames de histocompatibilidade HLA Paiva, Sérgio Ré de Sistema HLA Sorologia HLA Microlinfocitotoxicidade Tipagem HLA System HLA Serology Microiymphocytotoxicity HLA Typing |
title_short |
Técnica alternativa para fechamento de placas de microtécnica em exames de histocompatibilidade HLA |
title_full |
Técnica alternativa para fechamento de placas de microtécnica em exames de histocompatibilidade HLA |
title_fullStr |
Técnica alternativa para fechamento de placas de microtécnica em exames de histocompatibilidade HLA |
title_full_unstemmed |
Técnica alternativa para fechamento de placas de microtécnica em exames de histocompatibilidade HLA |
title_sort |
Técnica alternativa para fechamento de placas de microtécnica em exames de histocompatibilidade HLA |
author |
Paiva, Sérgio Ré de |
author_facet |
Paiva, Sérgio Ré de Paiva, Tádia Maria Cordeiro de Pontes, Luciane Faria de Souza Morínigo, Fábio Cupertino |
author_role |
author |
author2 |
Paiva, Tádia Maria Cordeiro de Pontes, Luciane Faria de Souza Morínigo, Fábio Cupertino |
author2_role |
author author author |
dc.contributor.author.fl_str_mv |
Paiva, Sérgio Ré de Paiva, Tádia Maria Cordeiro de Pontes, Luciane Faria de Souza Morínigo, Fábio Cupertino |
dc.subject.por.fl_str_mv |
Sistema HLA Sorologia HLA Microlinfocitotoxicidade Tipagem HLA System HLA Serology Microiymphocytotoxicity HLA Typing |
topic |
Sistema HLA Sorologia HLA Microlinfocitotoxicidade Tipagem HLA System HLA Serology Microiymphocytotoxicity HLA Typing |
description |
The HLA system consists in a group of cell membrane proteins: the HLA-A, B, C, DR, DQ and DP molecules. Arnong the main uses of this system we can find the farnily and random studies for bone marrow trans plantation, as well as the cross-matches before kidney transpiantations. An irnportant rnethodological advance in the study of the HLA system arised very soon, with the development of a rnicrolyrnphocytotoxiciiy technique, used until nowadays, in which the mononuclear blood cells bearing the HLA molecules are used as target for cytotoxicity reactions developed in specialized rnicrotrays. The final step of the serological rnethod consists to cover the tray with a glass cover slip and to read the reactions at an inverted light microscope. We present an alternative technique to dose the tray, which perrníts the utilization of a common light microscope. In this proposed method, a gel (Kaiser's rnodífied gelatin) is placed gently over the tray at the end of the usual technique, following eosin staining and formaline fixation. After that, the tray is placed, in inverted position, at a common light microscope and read in the same way as the usual rnethod. Twenty preliminary tests were perforrned unti/ now, with a very good reprodutibilily in terrns of reaction patterns; no mor phological altera tions were seen in these tests, when cornpared with the conventional technique. The use of this gelatin proved to be econornical, easy to perforrn and very reproducible. The trays covered with the gelatin were read again each two weeks for six rnonths, and we did not find any difference in the reaction patterns as well as in the cell morphology. |
publishDate |
2022 |
dc.date.none.fl_str_mv |
2022-10-04 |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
https://rbc.inca.gov.br/index.php/revista/article/view/2958 10.32635/2176-9745.RBC.1995v41n4.2958 |
url |
https://rbc.inca.gov.br/index.php/revista/article/view/2958 |
identifier_str_mv |
10.32635/2176-9745.RBC.1995v41n4.2958 |
dc.language.iso.fl_str_mv |
por |
language |
por |
dc.relation.none.fl_str_mv |
https://rbc.inca.gov.br/index.php/revista/article/view/2958/1831 |
dc.rights.driver.fl_str_mv |
https://creativecommons.org/licenses/by/4.0 info:eu-repo/semantics/openAccess |
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https://creativecommons.org/licenses/by/4.0 |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
application/pdf |
dc.publisher.none.fl_str_mv |
INCA |
publisher.none.fl_str_mv |
INCA |
dc.source.none.fl_str_mv |
Revista Brasileira de Cancerologia; Vol. 41 No. 4 (1995): Oct./Nov./Dec.; 271-274 Revista Brasileira de Cancerologia; Vol. 41 Núm. 4 (1995): oct./nov./dic.; 271-274 Revista Brasileira de Cancerologia; v. 41 n. 4 (1995): out./nov./dez.; 271-274 2176-9745 reponame:Revista Brasileira de Cancerologia (Online) instname:Instituto Nacional de Câncer José Alencar Gomes da Silva (INCA) instacron:INCA |
instname_str |
Instituto Nacional de Câncer José Alencar Gomes da Silva (INCA) |
instacron_str |
INCA |
institution |
INCA |
reponame_str |
Revista Brasileira de Cancerologia (Online) |
collection |
Revista Brasileira de Cancerologia (Online) |
repository.name.fl_str_mv |
Revista Brasileira de Cancerologia (Online) - Instituto Nacional de Câncer José Alencar Gomes da Silva (INCA) |
repository.mail.fl_str_mv |
rbc@inca.gov.br |
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1797042233435750400 |