OBJECTIVE: Recent clinical trials of adoptive immunotherapy showed diminished reactivity of human T cells upon ex vivo manipulation. For a safe and effective clinical application of human T cells, it is necessary to improve ex vivo manipulation procedures and evaluate their impact on in vivo functionality. However, there is no preclinical model for quantitative assessment of in vivo functionality of human T cells. In this study, we investigated the feasibility of using the huPBMC- RAG2(-/-)gammac(-/-) xenogeneic mouse model. As a first example, we compared 3 different ex vivo culture conditions for human T cells. METHODS: RAG2(-/-)gammac(-/-) mice received cultured human T cells that were stimulated via CD3 alone or costimulated via CD28 (CD3/28) and/or human 4-1BB (CD3/28/4-1BB). Engraftment levels and survival of the cells were measured. The dynamics of the human T cell phenotypes were analyzed during culture and in vivo, as well as the mechanism of the xenoresponse. RESULTS: Engraftment potential was improved twofold for costimulation compared to CD3 alone (p < 0.001). Phenotypic analysis showed a strikingly similar pattern of development towards CD4(+) and CD8(+) effector and effector-memory cells, suggesting antigen-driven survival and expansion. All parameters used to analyze different effects on in vivo T-cell functionality, like culture condition, engraftment levels, survival of the cells over time, or xenogeneic graft-vs-host disease were absolutely independent of the distribution of the T cell population in vivo following contact with xeno-antigen. CONCLUSION: The huPBMC-RAG2(-/-)gammac(-/-) xenogeneic transplant model is the most sensitive to date for in vivo functional evaluation of human T cells.
OBJECTIVE: Recent clinical trials of adoptive immunotherapy showed diminished reactivity of human T cells upon ex vivo manipulation. For a safe and effective clinical application of human T cells, it is necessary to improve ex vivo manipulation procedures and evaluate their impact on in vivo functionality. However, there is no preclinical model for quantitative assessment of in vivo functionality of human T cells. In this study, we investigated the feasibility of using the huPBMC- RAG2(-/-)gammac(-/-) xenogeneic mouse model. As a first example, we compared 3 different ex vivo culture conditions for human T cells. METHODS:RAG2(-/-)gammac(-/-) mice received cultured human T cells that were stimulated via CD3 alone or costimulated via CD28 (CD3/28) and/or human4-1BB (CD3/28/4-1BB). Engraftment levels and survival of the cells were measured. The dynamics of the human T cell phenotypes were analyzed during culture and in vivo, as well as the mechanism of the xenoresponse. RESULTS: Engraftment potential was improved twofold for costimulation compared to CD3 alone (p < 0.001). Phenotypic analysis showed a strikingly similar pattern of development towards CD4(+) and CD8(+) effector and effector-memory cells, suggesting antigen-driven survival and expansion. All parameters used to analyze different effects on in vivo T-cell functionality, like culture condition, engraftment levels, survival of the cells over time, or xenogeneic graft-vs-host disease were absolutely independent of the distribution of the T cell population in vivo following contact with xeno-antigen. CONCLUSION: The huPBMC-RAG2(-/-)gammac(-/-) xenogeneic transplant model is the most sensitive to date for in vivo functional evaluation of human T cells.
Authors: Céline Leboeuf; Laurent Mailly; Tao Wu; Gaetan Bour; Sarah Durand; Nicolas Brignon; Christophe Ferrand; Christophe Borg; Pierre Tiberghien; Robert Thimme; Patrick Pessaux; Jacques Marescaux; Thomas F Baumert; Eric Robinet Journal: Mol Ther Date: 2013-12-06 Impact factor: 11.454
Authors: Shoba Amarnath; Jason E Foley; Don E Farthing; Ronald E Gress; Arian Laurence; Michael A Eckhaus; Jean-Yves Métais; Jeremy J Rose; Frances T Hakim; Tania C Felizardo; Austin V Cheng; Pamela G Robey; David E Stroncek; Marianna Sabatino; Minoo Battiwalla; Sawa Ito; Daniel H Fowler; Austin J Barrett Journal: Stem Cells Date: 2015-04 Impact factor: 6.277
Authors: Niwa Ali; Barry Flutter; Robert Sanchez Rodriguez; Ehsan Sharif-Paghaleh; Linda D Barber; Giovanna Lombardi; Frank O Nestle Journal: PLoS One Date: 2012-08-28 Impact factor: 3.240
Authors: M A King; L Covassin; M A Brehm; W Racki; T Pearson; J Leif; J Laning; W Fodor; O Foreman; L Burzenski; T H Chase; B Gott; A A Rossini; R Bortell; L D Shultz; D L Greiner Journal: Clin Exp Immunol Date: 2009-07 Impact factor: 4.330
Authors: Ilaria Nichele; Alberto Zamò; Anna Bertolaso; Francesco Bifari; Martina Tinelli; Marta Franchini; Roberta Stradoni; Fiorenza Aprili; Giovanni Pizzolo; Mauro Krampera Journal: PLoS One Date: 2012-12-21 Impact factor: 3.240