title: Comparative and functional evaluation of in vitro generated to ex vivo CD8 T cells.
abstract: The generation of the cytotoxic CD8 T cell response is dependent on the functional outcomes imposed by the intrathymic constraints of differentiation and self-tolerance. Although thymic function can be partly replicated in vitro using OP9-DL1 cell cultures to yield CD8 αβ TCR-bearing cells from hematopoietic progenitor cells, a comprehensive and functional assessment of entirely in vitro generated CD8 T cells derived from bone marrow hematopoietic stem cells has not been established and remains controversial. In this study, we demonstrate that a phenotypic, molecular, and functional signature of in vitro derived CD8 T cells is akin to that of ex vivo CD8 T cells, although several significant differences were also observed. Transfer of in vitro derived CD8 T cells into syngeneic and immunodeficient host mice showed no graft-versus-host response, whereas a robust homeostatic proliferation was observed, respectively. These findings, along with a diverse and broad TCR repertoire expressed by the in vitro derived CD8 T cells, allowed for the successful generation of Ag-specific T cells to be obtained from an entirely in vitro generated CD8 T cell pool. These findings support the use of Ag-specific in vitro derived effector CD8 T cells for immune reconstitution approaches, which would be amenable to further tailoring for their use against viral infections or malignancies.
authors.affiliation: Department of Immunology, University of Toronto, Toronto, Ontario M4N 3M5, Canada.
authors.forename: Dzana D
authors.initials: DD
authors.lastname: Dervovic
authors.forename: Maria
authors.initials: M
authors.lastname: Ciofani
authors.forename: Korosh
authors.initials: K
authors.lastname: Kianizad
authors.forename: Juan Carlos
authors.initials: JC
authors.lastname: Zúñiga-Pflücker
mesh_terms: D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology / Q000276:immunology / Q000378:metabolism; D018414:CD8-Positive T-Lymphocytes / Q000166:cytology* / Q000276:immunology* / Q000378:metabolism; D002454:Cell Differentiation / Q000276:immunology*; D002460:Cell Line; D002478:Cells, Cultured; D016922:Cellular Senescence / Q000276:immunology; D018920:Coculture Techniques; D003601:Cytotoxicity Tests, Immunologic / Q000379:methods; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000276:immunology / Q000378:metabolism; D016130:Immunophenotyping / Q000379:methods; D051379:Mice; D020297:Mice, Congenic; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D011948:Receptors, Antigen, T-Cell / Q000096:biosynthesis / Q000502:physiology; D017154:Stromal Cells / Q000276:immunology / Q000378:metabolism
chemical_list: D011948:Receptors, Antigen, T-Cell