High Efficiency Ex Vivo Cloning of Antigen-Specific Human Effector T Cells

Michelle A. Neller, Michael H. -L. Lai, Catherine M. Lanagan, Linda E. O'Connor, Antonia L. Pritchard, Nathan R. Martinez, Christopher W. Schmidt

Research output: Contribution to journalArticlepeer-review

3 Citations (Scopus)


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.
Original languageEnglish
Pages (from-to)1-10
Number of pages10
JournalPLoS ONE
Issue number11
Publication statusPublished - 4 Nov 2014


Dive into the research topics of 'High Efficiency Ex Vivo Cloning of Antigen-Specific Human Effector T Cells'. Together they form a unique fingerprint.

Cite this