DNA-scaffolded biomaterials enable modular and tunable control of cell-based cancer immunotherapies

Xiao Huang; Jasper Z. Williams; Ryan Chang; Zhongbo Li; Eric Gai; David M. Patterson; Yu Wei; Wendell A. Lim; Tejal A. Desai

Anti-IL-2 clone #5355 (ThermoFisher #MA523696) was biotinylated using NHS- The copyright holder for this preprint (which was not peer-reviewed) is the author/funder. . https://doi.org/10.1101/587105 doi: bioRxiv preprint mCherry expression to easily identify transduced T cells 28 . All constructs were cloned via InFusion cloning (Takara Bio #638910). Pantropic VSV-G pseudotyped lentivirus was produced via transfection of Lenti-X 293T cells with a pHR'SIN:CSW transgene expression vector and the viral packaging plasmids pCMVdR8.91 and pMD2.G using Fugene HD (Promega #E2312). Primary T cells were thawed the same day and, after 24 hours in culture, were stimulated with Human T-Activator CD3/CD28 Dynabeads (Life Technologies #11131D) at a 1:2.5 cell:bead ratio. At 48 hours, viral supernatant was harvested and the primary T cells were exposed to the virus for 24 hours. At day 5 after T cell stimulation, Dynabeads were removed and the T cells expanded until day 12 when they were rested and could be used in assays. T cells were sorted for assays with a FACs ARIA II on day 5 post T cell stimulation.