Abstract
Determining the activity of a regulatory T-cell population in vitro is often the first step in analyzing its function. To obtain reliable and reproducible results, it is critical to follow the protocol that is most applicable to your experimental question. We have outlined below a basic in vitro suppression assay as well as a variety of alternative/additional protocols that can be utilized alone or in combination as desired.
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Acknowledgments
We wish to thank members of the Vignali lab for many discussions regarding these methods. We are particularly grateful to Andrea Szymczak-Workman (for advice on anti-CD3/CD28 bead conjugation), Creg Workman and Andrea Szymczak-Workman (set up of murine antigen specific suppression assays), Janice Riberdy (human suppression assay setup), and Sam Connell (CFSE labeling). LWC is supported by an Individual NIH NRSA (F32 AI072816). DAAV is supported by the National Institutes of Health (NIH) (AI39480, AI52199, AI072239), Juvenile Diabetes Research Foundation International (1-2004-141 [The Robert and Janice Compton Research Grant, In Honor of Elizabeth S. Compton] and 1-2006-847), a Cancer Center Support CORE grant (CA21765), and the American Lebanese Syrian Associated Charities (ALSAC).
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Collison, L.W., Vignali, D.A.A. (2011). In Vitro Treg Suppression Assays. In: Kassiotis, G., Liston, A. (eds) Regulatory T Cells. Methods in Molecular Biology, vol 707. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61737-979-6_2
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DOI: https://doi.org/10.1007/978-1-61737-979-6_2
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