High efficiency ex vivo cloning of antigen-specific human effector T cells.

While cloned T cells are valuable tools for the exploration of immune responses against viruses and tumours, current cloning methods do not allow inferences to be made about the function and phenotype of a clone's in vivo precursor, nor can precise cloning efficiencies be calculated. Additional...

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Autores principales:	Michelle A Neller, Michael H-L Lai, Catherine M Lanagan, Linda E O'Connor, Antonia L Pritchard, Nathan R Martinez, Christopher W Schmidt
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2014
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/d11adaf970244ecea3c97633192bcdc9
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spelling	oai:doaj.org-article:d11adaf970244ecea3c97633192bcdc92021-11-25T05:54:45ZHigh efficiency ex vivo cloning of antigen-specific human effector T cells.1932-620310.1371/journal.pone.0110741https://doaj.org/article/d11adaf970244ecea3c97633192bcdc92014-01-01T00:00:00Zhttps://doi.org/10.1371/journal.pone.0110741https://doaj.org/toc/1932-6203While cloned T cells are valuable tools for the exploration of immune responses against viruses and tumours, current cloning methods do not allow inferences to be made about the function and phenotype of a clone's in vivo precursor, nor can precise cloning efficiencies be calculated. Additionally, there is currently no general method for cloning antigen-specific effector T cells directly from peripheral blood mononuclear cells, without the need for prior expansion in vitro. Here we describe an efficient method for cloning effector T cells ex vivo. Functional T cells are detected using optimised interferon gamma capture following stimulation with viral or tumour cell-derived antigen. In combination with multiple phenotypic markers, single effector T cells are sorted using a flow cytometer directly into multi-well plates, and cloned using standard, non antigen-specific expansion methods. We provide examples of this novel technology to generate antigen-reactive clones from healthy donors using Epstein-Barr virus and cytomegalovirus as representative viral antigen sources, and from two melanoma patients using autologous melanoma cells. Cloning efficiency, clonality, and retention/loss of function are described. Ex vivo effector cell cloning provides a rapid and effective method of deriving antigen-specific T cells clones with traceable in vivo precursor function and phenotype.Michelle A NellerMichael H-L LaiCatherine M LanaganLinda E O'ConnorAntonia L PritchardNathan R MartinezChristopher W SchmidtPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 9, Iss 11, p e110741 (2014)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Michelle A Neller Michael H-L Lai Catherine M Lanagan Linda E O'Connor Antonia L Pritchard Nathan R Martinez Christopher W Schmidt High efficiency ex vivo cloning of antigen-specific human effector T cells.
description	While cloned T cells are valuable tools for the exploration of immune responses against viruses and tumours, current cloning methods do not allow inferences to be made about the function and phenotype of a clone's in vivo precursor, nor can precise cloning efficiencies be calculated. Additionally, there is currently no general method for cloning antigen-specific effector T cells directly from peripheral blood mononuclear cells, without the need for prior expansion in vitro. Here we describe an efficient method for cloning effector T cells ex vivo. Functional T cells are detected using optimised interferon gamma capture following stimulation with viral or tumour cell-derived antigen. In combination with multiple phenotypic markers, single effector T cells are sorted using a flow cytometer directly into multi-well plates, and cloned using standard, non antigen-specific expansion methods. We provide examples of this novel technology to generate antigen-reactive clones from healthy donors using Epstein-Barr virus and cytomegalovirus as representative viral antigen sources, and from two melanoma patients using autologous melanoma cells. Cloning efficiency, clonality, and retention/loss of function are described. Ex vivo effector cell cloning provides a rapid and effective method of deriving antigen-specific T cells clones with traceable in vivo precursor function and phenotype.
format	article
author	Michelle A Neller Michael H-L Lai Catherine M Lanagan Linda E O'Connor Antonia L Pritchard Nathan R Martinez Christopher W Schmidt
author_facet	Michelle A Neller Michael H-L Lai Catherine M Lanagan Linda E O'Connor Antonia L Pritchard Nathan R Martinez Christopher W Schmidt
author_sort	Michelle A Neller
title	High efficiency ex vivo cloning of antigen-specific human effector T cells.
title_short	High efficiency ex vivo cloning of antigen-specific human effector T cells.
title_full	High efficiency ex vivo cloning of antigen-specific human effector T cells.
title_fullStr	High efficiency ex vivo cloning of antigen-specific human effector T cells.
title_full_unstemmed	High efficiency ex vivo cloning of antigen-specific human effector T cells.
title_sort	high efficiency ex vivo cloning of antigen-specific human effector t cells.
publisher	Public Library of Science (PLoS)
publishDate	2014
url	https://doaj.org/article/d11adaf970244ecea3c97633192bcdc9
work_keys_str_mv	AT michelleaneller highefficiencyexvivocloningofantigenspecifichumaneffectortcells AT michaelhllai highefficiencyexvivocloningofantigenspecifichumaneffectortcells AT catherinemlanagan highefficiencyexvivocloningofantigenspecifichumaneffectortcells AT lindaeoconnor highefficiencyexvivocloningofantigenspecifichumaneffectortcells AT antonialpritchard highefficiencyexvivocloningofantigenspecifichumaneffectortcells AT nathanrmartinez highefficiencyexvivocloningofantigenspecifichumaneffectortcells AT christopherwschmidt highefficiencyexvivocloningofantigenspecifichumaneffectortcells
_version_	1718414426915209216

High efficiency ex vivo cloning of antigen-specific human effector T cells.

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