Avaliação comparativa das proteinas de fusão cmx e ecmx no teste de mantoux para o diagnóstico de tuberculose
Autor(a) principal: | |
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Data de Publicação: | 2017 |
Tipo de documento: | Dissertação |
Idioma: | por |
Título da fonte: | Repositório Institucional da UFG |
dARK ID: | ark:/38995/0013000002tf8 |
Texto Completo: | http://repositorio.bc.ufg.br/tede/handle/tede/7004 |
Resumo: | Tuberculin skin test (TST) identifies a previous exposed to M. tuberculosis (Mtb) using an intradermal inoculation of purified protein derivates (PPD) that result in a delayed hypersensitivity reaction (DTH). Interferon-gamma release assay (IGRA) was suggested to replace TST. The IGRA uses antigens, ESAT-6 and CFP-10, absent in all BCG strains and some non tuberculous mycobacteria (NTM). However, reproducibility and high cost were limitations for endemic countries. For this reason, the development of new diagnose test for latent TB is necessary. Fusion proteins developed by our group has been recognized by the immune response generated by the infection with Mycobacterium tuberculosis. Thus the aim of this work was to evaluate the capacity of CMX or ECMX to be used in a Skin test for tuberculosis. BALB/c mice infected with Mtb were euthanized forty-five days after infection. Spleens, lungs and draining lymph nodes of infected mice were processed and evaluated by flow cytometry Both CD4 and CD8 IFN+ cells were able to recognize rCMX and rECMX. The skin test followed an evaluation of thickness/swelling ≥ PPD 2UT (positive control) to consider positive DTH. Based on thickness, at 24 h, rCMX 25μg (0.37±0.02) and rECMX 15-25μg (0.38±0.03/0,62±0,12) induced a positive DTH response. At 48h, rCMX 25μg (0.28±0.03) and rECMX 25μg (0.5±0.04) induced also a positive DTH reaction. In conclusion, fusion proteins rCMX and rECMX are recognized by infected mice with Mtb and skin test using rECMX 25μg induced better DTH response that of conventional PPD. |
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Junqueira-Kipnis, Ana Paulahttp://lattes.cnpq.br/1252262903952987Junqueira-Kipnis, Ana PaulaSouza, Menira Borges de Lima Dias eBührer-Sékula, Samirahttp://lattes.cnpq.br/4243208739191599Sánchez, Tatiana Marlene Galvez2017-03-24T12:39:25Z2017-02-20Sánchez, T. M. G. Avaliação comparativa das proteinas de fusão cmx e ecmx no teste de mantoux para o diagnóstico de tuberculose. 2017. 74 f. Dissertação (Mestrado em Medicina Tropical e Saúde Publica) - Universidade Federal de Goiás, Goiânia, 2017.http://repositorio.bc.ufg.br/tede/handle/tede/7004ark:/38995/0013000002tf8Tuberculin skin test (TST) identifies a previous exposed to M. tuberculosis (Mtb) using an intradermal inoculation of purified protein derivates (PPD) that result in a delayed hypersensitivity reaction (DTH). Interferon-gamma release assay (IGRA) was suggested to replace TST. The IGRA uses antigens, ESAT-6 and CFP-10, absent in all BCG strains and some non tuberculous mycobacteria (NTM). However, reproducibility and high cost were limitations for endemic countries. For this reason, the development of new diagnose test for latent TB is necessary. Fusion proteins developed by our group has been recognized by the immune response generated by the infection with Mycobacterium tuberculosis. Thus the aim of this work was to evaluate the capacity of CMX or ECMX to be used in a Skin test for tuberculosis. BALB/c mice infected with Mtb were euthanized forty-five days after infection. Spleens, lungs and draining lymph nodes of infected mice were processed and evaluated by flow cytometry Both CD4 and CD8 IFN+ cells were able to recognize rCMX and rECMX. The skin test followed an evaluation of thickness/swelling ≥ PPD 2UT (positive control) to consider positive DTH. Based on thickness, at 24 h, rCMX 25μg (0.37±0.02) and rECMX 15-25μg (0.38±0.03/0,62±0,12) induced a positive DTH response. At 48h, rCMX 25μg (0.28±0.03) and rECMX 25μg (0.5±0.04) induced also a positive DTH reaction. In conclusion, fusion proteins rCMX and rECMX are recognized by infected mice with Mtb and skin test using rECMX 25μg induced better DTH response that of conventional PPD.A prova tuberculínica (PT) é um teste cutâneo que identifica a exposição prévia ao M. tuberculosis (Mtb), mediante a inoculação via intradérmica do derivado protéico purificado (PPD) de Mtb, o que resulta em uma reação de hipersensibilidade do tipo tardia (DTH). O ensaio de liberação de IFN-γ (IGRA) foi indicado para substituir a PT. O IGRA usa os antígenos ausentes na BCG e algumas micobactérias não causadoras de TB (MNT), ESAT-6 e CFP-10. Porém, apresenta falta de reprodutibilidade e alto custo quando usado em populações endêmicas para TB. Diante disso, o desenvolvimento de novos testes de diagnóstico é necessário. Nosso grupo desenvolveu proteínas de fusão que são reconhecidas por linfócitos gerados pela infecção com Mtb. Assim, o trabalho propõe avaliar a utilização das proteínas rCMX e rECMX no desenvolvimento de um teste cutâneo de diagnóstico para tuberculose. Camundongos BALB/c foram infectados com Mtb H37Rv. Após 45 dias, a infecção induziu linfócitos T CD4+ e CD8+ produtores de IFN-γ específicos para rCMX e rECMX no baço, pulmões e linfonodos drenantes. Enquanto ao teste cutâneo realizado 45 dias após a infecção, a leitura de espessura/inchaço ≥ PPD 2UT (controle positivo) indicou uma reação de DTH positiva. Avaliando a espessura 24h após o inóculo, rCMX 25μg (0.37±0.02) e rECMX 15-25μg (0.38±0.03/0,62±0,12) induziram reação de DTH positiva. As 48h, rCMX 25μg (0.28±0.03) e rECMX 25μg (0.5±0.04) também apresentaram reação positiva. Enquanto o inchaço as 24h, só a rECMX apresentou DTH positiva. Em conclusão, este trabalho mostra que as proteínas rCMX e rECMX são reconhecidas pela resposta celular de camundongos infectados com Mtb, e quando usadas no teste cutâneo induziram reação de DTH positiva comparável e até superior ao PPD convencional. Dessa forma, é recomendada a avaliação das proteínas de fusão em outros modelos animais e posteriormente em humanos.Submitted by Cássia Santos (cassia.bcufg@gmail.com) on 2017-03-24T11:22:47Z No. of bitstreams: 2 Dissertação - Tatiana Marlene Galvez Sánchez - 2017.pdf: 3124839 bytes, checksum: 942aa6d4fa9aa30babb4eb0d7e04a805 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5)Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2017-03-24T12:39:25Z (GMT) No. of bitstreams: 2 Dissertação - Tatiana Marlene Galvez Sánchez - 2017.pdf: 3124839 bytes, checksum: 942aa6d4fa9aa30babb4eb0d7e04a805 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5)Made available in DSpace on 2017-03-24T12:39:25Z (GMT). No. of bitstreams: 2 Dissertação - Tatiana Marlene Galvez Sánchez - 2017.pdf: 3124839 bytes, checksum: 942aa6d4fa9aa30babb4eb0d7e04a805 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2017-02-20Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPESOutroapplication/pdfhttp://repositorio.bc.ufg.br/tede/retrieve/37341/Disserta%c3%a7%c3%a3o%20-%20Tatiana%20Marlene%20Galvez%20S%c3%a1nchez%20-%202017.pdf.jpgporUniversidade Federal de GoiásPrograma de Pós-graduação em Medicina Tropical e Saúde Publica (IPTSP)UFGBrasilInstituto de Patologia Tropical e Saúde Pública - IPTSP (RG)http://creativecommons.org/licenses/by-nc-nd/4.0/info:eu-repo/semantics/openAccessTuberculose (TB)Prova tuberculínica (PT)Hipersensibilidade do tipo tardia (DTH)Tuberculosis (TB)Tuberculin skin test (TST)Delayed hypersensitivity reaction (DTH)CIENCIAS BIOLOGICAS::IMUNOLOGIAAvaliação comparativa das proteinas de fusão cmx e ecmx no teste de mantoux para o diagnóstico de tuberculoseComparative evaluation of fusion proteins cmx and ecmx by mantoux technique for tuberculosis diagnosisinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesis6085308344741430434600600600600600-7769011444564556288598991918837674761420751674985882645712442915598251853972reponame:Repositório Institucional da UFGinstname:Universidade Federal de Goiás (UFG)instacron:UFGCC-LICENSElicense_urllicense_urltext/plain; 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dc.title.por.fl_str_mv |
Avaliação comparativa das proteinas de fusão cmx e ecmx no teste de mantoux para o diagnóstico de tuberculose |
dc.title.alternative.eng.fl_str_mv |
Comparative evaluation of fusion proteins cmx and ecmx by mantoux technique for tuberculosis diagnosis |
title |
Avaliação comparativa das proteinas de fusão cmx e ecmx no teste de mantoux para o diagnóstico de tuberculose |
spellingShingle |
Avaliação comparativa das proteinas de fusão cmx e ecmx no teste de mantoux para o diagnóstico de tuberculose Sánchez, Tatiana Marlene Galvez Tuberculose (TB) Prova tuberculínica (PT) Hipersensibilidade do tipo tardia (DTH) Tuberculosis (TB) Tuberculin skin test (TST) Delayed hypersensitivity reaction (DTH) CIENCIAS BIOLOGICAS::IMUNOLOGIA |
title_short |
Avaliação comparativa das proteinas de fusão cmx e ecmx no teste de mantoux para o diagnóstico de tuberculose |
title_full |
Avaliação comparativa das proteinas de fusão cmx e ecmx no teste de mantoux para o diagnóstico de tuberculose |
title_fullStr |
Avaliação comparativa das proteinas de fusão cmx e ecmx no teste de mantoux para o diagnóstico de tuberculose |
title_full_unstemmed |
Avaliação comparativa das proteinas de fusão cmx e ecmx no teste de mantoux para o diagnóstico de tuberculose |
title_sort |
Avaliação comparativa das proteinas de fusão cmx e ecmx no teste de mantoux para o diagnóstico de tuberculose |
author |
Sánchez, Tatiana Marlene Galvez |
author_facet |
Sánchez, Tatiana Marlene Galvez |
author_role |
author |
dc.contributor.advisor1.fl_str_mv |
Junqueira-Kipnis, Ana Paula |
dc.contributor.advisor1Lattes.fl_str_mv |
http://lattes.cnpq.br/1252262903952987 |
dc.contributor.referee1.fl_str_mv |
Junqueira-Kipnis, Ana Paula |
dc.contributor.referee2.fl_str_mv |
Souza, Menira Borges de Lima Dias e |
dc.contributor.referee3.fl_str_mv |
Bührer-Sékula, Samira |
dc.contributor.authorLattes.fl_str_mv |
http://lattes.cnpq.br/4243208739191599 |
dc.contributor.author.fl_str_mv |
Sánchez, Tatiana Marlene Galvez |
contributor_str_mv |
Junqueira-Kipnis, Ana Paula Junqueira-Kipnis, Ana Paula Souza, Menira Borges de Lima Dias e Bührer-Sékula, Samira |
dc.subject.por.fl_str_mv |
Tuberculose (TB) Prova tuberculínica (PT) Hipersensibilidade do tipo tardia (DTH) |
topic |
Tuberculose (TB) Prova tuberculínica (PT) Hipersensibilidade do tipo tardia (DTH) Tuberculosis (TB) Tuberculin skin test (TST) Delayed hypersensitivity reaction (DTH) CIENCIAS BIOLOGICAS::IMUNOLOGIA |
dc.subject.eng.fl_str_mv |
Tuberculosis (TB) Tuberculin skin test (TST) Delayed hypersensitivity reaction (DTH) |
dc.subject.cnpq.fl_str_mv |
CIENCIAS BIOLOGICAS::IMUNOLOGIA |
description |
Tuberculin skin test (TST) identifies a previous exposed to M. tuberculosis (Mtb) using an intradermal inoculation of purified protein derivates (PPD) that result in a delayed hypersensitivity reaction (DTH). Interferon-gamma release assay (IGRA) was suggested to replace TST. The IGRA uses antigens, ESAT-6 and CFP-10, absent in all BCG strains and some non tuberculous mycobacteria (NTM). However, reproducibility and high cost were limitations for endemic countries. For this reason, the development of new diagnose test for latent TB is necessary. Fusion proteins developed by our group has been recognized by the immune response generated by the infection with Mycobacterium tuberculosis. Thus the aim of this work was to evaluate the capacity of CMX or ECMX to be used in a Skin test for tuberculosis. BALB/c mice infected with Mtb were euthanized forty-five days after infection. Spleens, lungs and draining lymph nodes of infected mice were processed and evaluated by flow cytometry Both CD4 and CD8 IFN+ cells were able to recognize rCMX and rECMX. The skin test followed an evaluation of thickness/swelling ≥ PPD 2UT (positive control) to consider positive DTH. Based on thickness, at 24 h, rCMX 25μg (0.37±0.02) and rECMX 15-25μg (0.38±0.03/0,62±0,12) induced a positive DTH response. At 48h, rCMX 25μg (0.28±0.03) and rECMX 25μg (0.5±0.04) induced also a positive DTH reaction. In conclusion, fusion proteins rCMX and rECMX are recognized by infected mice with Mtb and skin test using rECMX 25μg induced better DTH response that of conventional PPD. |
publishDate |
2017 |
dc.date.accessioned.fl_str_mv |
2017-03-24T12:39:25Z |
dc.date.issued.fl_str_mv |
2017-02-20 |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/masterThesis |
format |
masterThesis |
status_str |
publishedVersion |
dc.identifier.citation.fl_str_mv |
Sánchez, T. M. G. Avaliação comparativa das proteinas de fusão cmx e ecmx no teste de mantoux para o diagnóstico de tuberculose. 2017. 74 f. Dissertação (Mestrado em Medicina Tropical e Saúde Publica) - Universidade Federal de Goiás, Goiânia, 2017. |
dc.identifier.uri.fl_str_mv |
http://repositorio.bc.ufg.br/tede/handle/tede/7004 |
dc.identifier.dark.fl_str_mv |
ark:/38995/0013000002tf8 |
identifier_str_mv |
Sánchez, T. M. G. Avaliação comparativa das proteinas de fusão cmx e ecmx no teste de mantoux para o diagnóstico de tuberculose. 2017. 74 f. Dissertação (Mestrado em Medicina Tropical e Saúde Publica) - Universidade Federal de Goiás, Goiânia, 2017. ark:/38995/0013000002tf8 |
url |
http://repositorio.bc.ufg.br/tede/handle/tede/7004 |
dc.language.iso.fl_str_mv |
por |
language |
por |
dc.relation.program.fl_str_mv |
6085308344741430434 |
dc.relation.confidence.fl_str_mv |
600 600 600 600 600 |
dc.relation.department.fl_str_mv |
-7769011444564556288 |
dc.relation.cnpq.fl_str_mv |
5989919188376747614 |
dc.relation.sponsorship.fl_str_mv |
2075167498588264571 2442915598251853972 |
dc.rights.driver.fl_str_mv |
http://creativecommons.org/licenses/by-nc-nd/4.0/ info:eu-repo/semantics/openAccess |
rights_invalid_str_mv |
http://creativecommons.org/licenses/by-nc-nd/4.0/ |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
application/pdf |
dc.publisher.none.fl_str_mv |
Universidade Federal de Goiás |
dc.publisher.program.fl_str_mv |
Programa de Pós-graduação em Medicina Tropical e Saúde Publica (IPTSP) |
dc.publisher.initials.fl_str_mv |
UFG |
dc.publisher.country.fl_str_mv |
Brasil |
dc.publisher.department.fl_str_mv |
Instituto de Patologia Tropical e Saúde Pública - IPTSP (RG) |
publisher.none.fl_str_mv |
Universidade Federal de Goiás |
dc.source.none.fl_str_mv |
reponame:Repositório Institucional da UFG instname:Universidade Federal de Goiás (UFG) instacron:UFG |
instname_str |
Universidade Federal de Goiás (UFG) |
instacron_str |
UFG |
institution |
UFG |
reponame_str |
Repositório Institucional da UFG |
collection |
Repositório Institucional da UFG |
bitstream.url.fl_str_mv |
http://repositorio.bc.ufg.br/tede/bitstreams/7c96c830-5a53-432d-8cc7-5a9683c73894/download http://repositorio.bc.ufg.br/tede/bitstreams/b11778b0-a043-4b7f-a32e-46db451c3c08/download http://repositorio.bc.ufg.br/tede/bitstreams/527a5113-f4d5-4fc1-9f9a-7746f8a7623e/download http://repositorio.bc.ufg.br/tede/bitstreams/31910a56-cd78-4275-8384-5918519a96e8/download http://repositorio.bc.ufg.br/tede/bitstreams/0c7e267d-8859-4e01-8e82-8184297496c2/download http://repositorio.bc.ufg.br/tede/bitstreams/b0dfc8ed-d7f6-436e-a5ab-76d843e3bcb2/download http://repositorio.bc.ufg.br/tede/bitstreams/0540270d-cda4-4c10-98ad-79784ea3287f/download |
bitstream.checksum.fl_str_mv |
4afdbb8c545fd630ea7db775da747b2f d41d8cd98f00b204e9800998ecf8427e d41d8cd98f00b204e9800998ecf8427e 942aa6d4fa9aa30babb4eb0d7e04a805 bd3efa91386c1718a7f26a329fdcb468 57d616707843d3514863a4c95972c082 0ac13f55c762e8980218cec15e7adaff |
bitstream.checksumAlgorithm.fl_str_mv |
MD5 MD5 MD5 MD5 MD5 MD5 MD5 |
repository.name.fl_str_mv |
Repositório Institucional da UFG - Universidade Federal de Goiás (UFG) |
repository.mail.fl_str_mv |
tasesdissertacoes.bc@ufg.br |
_version_ |
1815172538916929536 |