Abstract
T regulatory cells (Tregs) suppress immune responses and therefore have potential to be used in the clinic as a cellular therapy for autoimmune disease and to prevent rejection of transplanted organs. Obtaining sufficient numbers of these cells for therapeutic use is a challenge, however, since there are currently no Treg cell-specific markers, and they have a poor in vitro expansion potential. Tregs express high levels of FOXP3, a transcription factor that is critical for their function. We have shown that lentivirus-based overexpression of FOXP3 can reprogram naïve or memory human CD4+ T cells into cells which possess a phenotype and function similar to ex vivo Tregs. Here we will review the methodology involved in generating, expanding, and testing FOXP3-transduced cells and their ex vivo Treg counterparts.
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Acknowledgments
We thank Sarah E. Allan, Rosa Bacchetta, Maria Grazia Roncarolo, Mario Amendola, and Luigi Naldini for their contributions to the development of this protocol. Supported by the Roche Organ Transplant Research Foundation, CIHR (MOP-93793), and Stemcell Technologies, Inc. Core support for flow cytometry and virus production was funded by the Immunity and Infection Research Centre Michael Smith Foundation for Health Research (MSFHR) Unit. MKL holds a Canada Research Chair in Transplantation. ANM holds a Canada Vanier Scholarship, a MSFHR Junior Graduate Studentship, and a CIHR Transplantation Training Program award.
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McMurchy, A.N., Levings, M.K. (2013). In Vitro Generation of Human T Regulatory Cells: Generation, Culture, and Analysis of FOXP3-Transduced T Cells. In: Helgason, C., Miller, C. (eds) Basic Cell Culture Protocols. Methods in Molecular Biology, vol 946. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-128-8_8
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DOI: https://doi.org/10.1007/978-1-62703-128-8_8
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