Evaluation of approaches to identify the targets of cellular immunity on a proteome-wide scale.

<h4>Background</h4>Vaccine development against malaria and other complex diseases remains a challenge for the scientific community. The recent elucidation of the genome, proteome and transcriptome of many of these complex pathogens provides the basis for rational vaccine design by identi...

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Autores principales:	Fernanda C Cardoso, Joanne S Roddick, Penny Groves, Denise L Doolan
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2011
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/3d934c405da34b8a9af8fe2efbba4415
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spelling	oai:doaj.org-article:3d934c405da34b8a9af8fe2efbba44152021-11-18T07:34:26ZEvaluation of approaches to identify the targets of cellular immunity on a proteome-wide scale.1932-620310.1371/journal.pone.0027666https://doaj.org/article/3d934c405da34b8a9af8fe2efbba44152011-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/22096610/pdf/?tool=EBIhttps://doaj.org/toc/1932-6203<h4>Background</h4>Vaccine development against malaria and other complex diseases remains a challenge for the scientific community. The recent elucidation of the genome, proteome and transcriptome of many of these complex pathogens provides the basis for rational vaccine design by identifying, on a proteome-wide scale, novel target antigens that are recognized by T cells and antibodies from exposed individuals. However, there is currently no algorithm to effectively identify important target antigens from genome sequence data; this is especially challenging for T cell targets. Furthermore, for some of these pathogens, such as Plasmodium, protein expression using conventional platforms has been problematic but cell-free in vitro transcription translation (IVTT) strategies have recently proved successful. Herein, we report a novel approach for proteome-wide scale identification of the antigenic targets of T cell responses using IVTT products.<h4>Principal findings</h4>We conducted a series of in vitro and in vivo experiments using IVTT proteins either unpurified, absorbed to carboxylated polybeads, or affinity purified through nickel resin or magnetic beads. In vitro studies in humans using CMV, EBV, and Influenza A virus proteins showed antigen-specific cytokine production in ELIspot and Cytometric Bead Array assays with cells stimulated with purified or unpurified IVTT antigens. In vitro and in vivo studies in mice immunized with the Plasmodium yoelii circumsporozoite DNA vaccine with or without IVTT protein boost showed antigen-specific cytokine production using purified IVTT antigens only. Overall, the nickel resin method of IVTT antigen purification proved optimal in both human and murine systems.<h4>Conclusions</h4>This work provides proof of concept for the potential of high-throughput approaches to identify T cell targets of complex parasitic, viral or bacterial pathogens from genomic sequence data, for rational vaccine development against emerging and re-emerging diseases that pose a threat to public health.Fernanda C CardosoJoanne S RoddickPenny GrovesDenise L DoolanPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 6, Iss 11, p e27666 (2011)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Fernanda C Cardoso Joanne S Roddick Penny Groves Denise L Doolan Evaluation of approaches to identify the targets of cellular immunity on a proteome-wide scale.
description	<h4>Background</h4>Vaccine development against malaria and other complex diseases remains a challenge for the scientific community. The recent elucidation of the genome, proteome and transcriptome of many of these complex pathogens provides the basis for rational vaccine design by identifying, on a proteome-wide scale, novel target antigens that are recognized by T cells and antibodies from exposed individuals. However, there is currently no algorithm to effectively identify important target antigens from genome sequence data; this is especially challenging for T cell targets. Furthermore, for some of these pathogens, such as Plasmodium, protein expression using conventional platforms has been problematic but cell-free in vitro transcription translation (IVTT) strategies have recently proved successful. Herein, we report a novel approach for proteome-wide scale identification of the antigenic targets of T cell responses using IVTT products.<h4>Principal findings</h4>We conducted a series of in vitro and in vivo experiments using IVTT proteins either unpurified, absorbed to carboxylated polybeads, or affinity purified through nickel resin or magnetic beads. In vitro studies in humans using CMV, EBV, and Influenza A virus proteins showed antigen-specific cytokine production in ELIspot and Cytometric Bead Array assays with cells stimulated with purified or unpurified IVTT antigens. In vitro and in vivo studies in mice immunized with the Plasmodium yoelii circumsporozoite DNA vaccine with or without IVTT protein boost showed antigen-specific cytokine production using purified IVTT antigens only. Overall, the nickel resin method of IVTT antigen purification proved optimal in both human and murine systems.<h4>Conclusions</h4>This work provides proof of concept for the potential of high-throughput approaches to identify T cell targets of complex parasitic, viral or bacterial pathogens from genomic sequence data, for rational vaccine development against emerging and re-emerging diseases that pose a threat to public health.
format	article
author	Fernanda C Cardoso Joanne S Roddick Penny Groves Denise L Doolan
author_facet	Fernanda C Cardoso Joanne S Roddick Penny Groves Denise L Doolan
author_sort	Fernanda C Cardoso
title	Evaluation of approaches to identify the targets of cellular immunity on a proteome-wide scale.
title_short	Evaluation of approaches to identify the targets of cellular immunity on a proteome-wide scale.
title_full	Evaluation of approaches to identify the targets of cellular immunity on a proteome-wide scale.
title_fullStr	Evaluation of approaches to identify the targets of cellular immunity on a proteome-wide scale.
title_full_unstemmed	Evaluation of approaches to identify the targets of cellular immunity on a proteome-wide scale.
title_sort	evaluation of approaches to identify the targets of cellular immunity on a proteome-wide scale.
publisher	Public Library of Science (PLoS)
publishDate	2011
url	https://doaj.org/article/3d934c405da34b8a9af8fe2efbba4415
work_keys_str_mv	AT fernandaccardoso evaluationofapproachestoidentifythetargetsofcellularimmunityonaproteomewidescale AT joannesroddick evaluationofapproachestoidentifythetargetsofcellularimmunityonaproteomewidescale AT pennygroves evaluationofapproachestoidentifythetargetsofcellularimmunityonaproteomewidescale AT deniseldoolan evaluationofapproachestoidentifythetargetsofcellularimmunityonaproteomewidescale
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Evaluation of approaches to identify the targets of cellular immunity on a proteome-wide scale.

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