Impairment of IFN-gamma response to synthetic peptides of mycobacterium tuberculosis in a 7-day whole blood assay

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dc.contributor.authorGideon, Hannah Priyadarshinien_ZA
dc.contributor.authorHamilton, Melissa Sheaen_ZA
dc.contributor.authorWood, Kathrynen_ZA
dc.contributor.authorPepper, Dominiqueen_ZA
dc.contributor.authorOni, Toluen_ZA
dc.contributor.authorSeldon, Ronnetten_ZA
dc.contributor.authorBanwell, Claireen_ZA
dc.contributor.authorLangford, Paul Ren_ZA
dc.contributor.authorWilkinson, Robert Jen_ZA
dc.contributor.authorWilkinson, Katalin Aen_ZA
dc.date.accessioned2015-12-28T06:47:18Z
dc.date.available2015-12-28T06:47:18Z
dc.date.issued2013en_ZA
dc.description.abstractStudies on Mycobacterium tuberculosis (MTB) antigens are of interest in order to improve vaccine efficacy and to define biomarkers for diagnosis and treatment monitoring. The methodologies used for these investigations differ greatly between laboratories and discordant results are common. The IFN-gamma response to two well characterized MTB antigens ESAT-6 and CFP-10, in the form of recombinant proteins and synthetic peptides, was evaluated in HIV-1 uninfected persons in both long-term (7 day) and 24 hour, commercially available QuantiFERON TB Gold in Tube (QFT-GIT), whole blood assays. Our findings showed differences in the IFN-gamma response between 24 hour and 7 day cultures, with recombinant proteins inducing a significantly higher response than the peptide pools in 7 day whole blood assays. The activity of peptides and recombinant proteins did not differ in 24 hour whole blood or peripheral blood mononuclear cell (PBMC) based assays, nor in the ELISpot assay. Further analysis by SELDI-TOF mass spectrometry showed that the peptides are degraded over the course of 7 days of incubation in whole blood whilst the recombinant proteins remain intact. This study therefore demonstrates that screening antigenic candidates as synthetic peptides in long-term whole blood assays may underestimate immunogenicity.en_ZA
dc.identifier.apacitationGideon, H. P., Hamilton, M. S., Wood, K., Pepper, D., Oni, T., Seldon, R., ... Wilkinson, K. A. (2013). Impairment of IFN-gamma response to synthetic peptides of mycobacterium tuberculosis in a 7-day whole blood assay. <i>PLoS One</i>, http://hdl.handle.net/11427/16022en_ZA
dc.identifier.chicagocitationGideon, Hannah Priyadarshini, Melissa Shea Hamilton, Kathryn Wood, Dominique Pepper, Tolu Oni, Ronnett Seldon, Claire Banwell, Paul R Langford, Robert J Wilkinson, and Katalin A Wilkinson "Impairment of IFN-gamma response to synthetic peptides of mycobacterium tuberculosis in a 7-day whole blood assay." <i>PLoS One</i> (2013) http://hdl.handle.net/11427/16022en_ZA
dc.identifier.citationGideon, H. P., Hamilton, M. S., Wood, K., Pepper, D., Oni, T., Seldon, R., ... & Wilkinson, K. A. (2013). Impairment of IFN-gamma response to synthetic peptides of mycobacterium tuberculosis in a 7-day whole blood assay. PloS one, 8(8), 71351. doi:10.1371/journal.pone.0071351en_ZA
dc.identifier.ris TY - Journal Article AU - Gideon, Hannah Priyadarshini AU - Hamilton, Melissa Shea AU - Wood, Kathryn AU - Pepper, Dominique AU - Oni, Tolu AU - Seldon, Ronnett AU - Banwell, Claire AU - Langford, Paul R AU - Wilkinson, Robert J AU - Wilkinson, Katalin A AB - Studies on Mycobacterium tuberculosis (MTB) antigens are of interest in order to improve vaccine efficacy and to define biomarkers for diagnosis and treatment monitoring. The methodologies used for these investigations differ greatly between laboratories and discordant results are common. The IFN-gamma response to two well characterized MTB antigens ESAT-6 and CFP-10, in the form of recombinant proteins and synthetic peptides, was evaluated in HIV-1 uninfected persons in both long-term (7 day) and 24 hour, commercially available QuantiFERON TB Gold in Tube (QFT-GIT), whole blood assays. Our findings showed differences in the IFN-gamma response between 24 hour and 7 day cultures, with recombinant proteins inducing a significantly higher response than the peptide pools in 7 day whole blood assays. The activity of peptides and recombinant proteins did not differ in 24 hour whole blood or peripheral blood mononuclear cell (PBMC) based assays, nor in the ELISpot assay. Further analysis by SELDI-TOF mass spectrometry showed that the peptides are degraded over the course of 7 days of incubation in whole blood whilst the recombinant proteins remain intact. This study therefore demonstrates that screening antigenic candidates as synthetic peptides in long-term whole blood assays may underestimate immunogenicity. DA - 2013 DB - OpenUCT DO - 10.1371/journal.pone.0071351 DP - University of Cape Town J1 - PLoS One LK - https://open.uct.ac.za PB - University of Cape Town PY - 2013 T1 - Impairment of IFN-gamma response to synthetic peptides of mycobacterium tuberculosis in a 7-day whole blood assay TI - Impairment of IFN-gamma response to synthetic peptides of mycobacterium tuberculosis in a 7-day whole blood assay UR - http://hdl.handle.net/11427/16022 ER - en_ZA
dc.identifier.urihttp://hdl.handle.net/11427/16022
dc.identifier.urihttp://dx.doi.org/10.1371/journal.pone.0071351
dc.identifier.vancouvercitationGideon HP, Hamilton MS, Wood K, Pepper D, Oni T, Seldon R, et al. Impairment of IFN-gamma response to synthetic peptides of mycobacterium tuberculosis in a 7-day whole blood assay. PLoS One. 2013; http://hdl.handle.net/11427/16022.en_ZA
dc.language.isoengen_ZA
dc.publisherPublic Library of Scienceen_ZA
dc.publisher.departmentInstitute of Infectious Disease and Molecular Medicineen_ZA
dc.publisher.facultyFaculty of Health Sciencesen_ZA
dc.publisher.institutionUniversity of Cape Town
dc.rightsThis is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.en_ZA
dc.rights.holder© 2013 Gideon et alen_ZA
dc.rights.urihttp://creativecommons.org/licenses/by/4.0en_ZA
dc.sourcePLoS Oneen_ZA
dc.source.urihttp://journals.plos.org/plosoneen_ZA
dc.subject.otherRecombinant proteinsen_ZA
dc.subject.otherBlooden_ZA
dc.subject.otherEnzyme-linked immunoassaysen_ZA
dc.subject.otherMycobacterium tuberculosisen_ZA
dc.subject.otherPeptidesen_ZA
dc.subject.otherSynthetic peptidesen_ZA
dc.subject.otherTuberculosisen_ZA
dc.subject.otherT cellsen_ZA
dc.titleImpairment of IFN-gamma response to synthetic peptides of mycobacterium tuberculosis in a 7-day whole blood assayen_ZA
dc.typeJournal Articleen_ZA
uct.type.filetypeText
uct.type.filetypeImage
uct.type.publicationResearchen_ZA
uct.type.resourceArticleen_ZA
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