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

Neller, Michelle A., Lai, Michael H. -L., Lanagan, Catherine M., O'Connor, Linda E., Pritchard, Antonia L., Martinez, Nathan R. and Schmidt, Christopher W. (2014) High efficiency ex vivo cloning of antigen-specific human effector T cells. Plos One, 9 11: . doi:10.1371/journal.pone.0110741


Author Neller, Michelle A.
Lai, Michael H. -L.
Lanagan, Catherine M.
O'Connor, Linda E.
Pritchard, Antonia L.
Martinez, Nathan R.
Schmidt, Christopher W.
Title High efficiency ex vivo cloning of antigen-specific human effector T cells
Formatted title
High efficiency ex vivo cloning of antigen-specific human effector T cells
Journal name Plos One   Check publisher's open access policy
ISSN 1932-6203
Publication date 2014-11
Year available 2014
Sub-type Article (original research)
DOI 10.1371/journal.pone.0110741
Open Access Status DOI
Volume 9
Issue 11
Total pages 10
Place of publication San Francisco United States
Publisher Public Library of Science (PLoS)
Collection year 2015
Language eng
Formatted abstract
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.
Keyword Human naive
In-vitro
Melanoma
Memory
Avidity
Q-Index Code C1
Q-Index Status Confirmed Code
Institutional Status UQ

Document type: Journal Article
Sub-type: Article (original research)
Collections: Official 2015 Collection
School of Medicine Publications
 
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