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- Title
High Efficiency Ex Vivo Cloning of Antigen-Specific Human Effector T Cells.
- Authors
Neller, Michelle A.; Lai, Michael H.-L.; Lanagan, Catherine M.; O′Connor, Linda E.; Pritchard, Antonia L.; Martinez, Nathan R.; Schmidt, Christopher W.
- 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.
- Subjects
T cells; IMMUNE response; CELLULAR immunity; PHENOTYPES; VIRAL antigens; TUMOR immunology
- Publication
PLoS ONE, 2014, Vol 9, Issue 11, p1
- ISSN
1932-6203
- Publication type
Article
- DOI
10.1371/journal.pone.0110741