Avaliação comparativa das proteinas de fusão cmx e ecmx no teste de mantoux para o diagnóstico de tuberculose

Detalhes bibliográficos
Autor(a) principal: Sánchez, Tatiana Marlene Galvez
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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spelling 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). 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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
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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.
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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
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dc.publisher.none.fl_str_mv Universidade Federal de Goiás
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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
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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
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repository.name.fl_str_mv Repositório Institucional da UFG - Universidade Federal de Goiás (UFG)
repository.mail.fl_str_mv tasesdissertacoes.bc@ufg.br
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