Correction to “Primary T-cell-based delivery platform for in vivo synthesis of engineered proteins”

Abstract

Radhakrishnan H, Newmyer SL, Ssemadaali MA, Javitz HS, Bhatnagar P. Primary T-cell-based delivery platform for in vivo synthesis of engineered proteins. Bioeng Transl Med. 2024; 9(1):e10605. doi:10.1002/btm2.10605

4.10 In vivo validation of delivery function of the engineered T cells (engineered for delivery function with NFAT-RE delivery system). The in vivo validation of our T-cell based delivery system was performed in mice at SRI International in accordance with the guidelines from the Institutional Animal Care and Use Committee (Approval # 22001). Six- to 8-week-old female NOD.Cg-Prkdc^scid Il2rg^tm1Wjl/SzJ (NSG) mice were purchased from The Jackson Laboratory. After mandatory quarantine, the NSG mice were anesthetized and 2 × 10⁶ FRα⁺Luc2-2A-E2Crimson⁺A2780cis cells in 100 μL 1× PBS were i.p. implanted. The tumor growth was monitored every 3–4 days for the next 12 days using i.p. injected 150 mg d-Luciferin per kg of mouse dissolved in 1× PBS. At 11 days after implantation, the mice were randomized into two groups (n = 5 each). The two groups were then treated with 2 × 10⁶ primary CD4 T cells engineered for delivery function (i.e., FRα-CAR with NFAT-RE inducible Nluc reporter) or the control primary CD4 T cells (FRα-CAR only, i.e., without NFAT-RE inducible Nluc reporter) every day for 5 days. The bioluminescent reporter (Nluc) activity was determined by i.p. injection of the Nano-Glo® substrate (1:20 dilution of the substrate in 1× PBS, equivalent to 0.5 mg per kg of mouse) on Days 0, 1, 2, 3, 4, and 5 after treatment. Imaging was performed in a IVIS Lumina X5 imaging system. The data were quantified by analysis of the ROI using Living Image software. The tumor luminescence is plotted as the mean ± SEM of total flux (photons/s) against days after treatment.