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. 2021 May 11;12:2637. doi: 10.1038/s41467-021-22898-3

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© The Author(s) 2021

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 1 — a EG7.OVA percent killing by RCT-activated OT-1 cells (n = 3). b CD45.1 Pep Boy mice were randomized (~180 mm³; n = 5), treated with naïve OT-1 cells, and dosed with 1 × 10⁹ mRBC-CTRL or mRBC-aAPC. c OT-1 number on days 0, 3, and 6. One-way ANOVA compared to mRBC-CTRL; Day 3: mRBC-OVA-4-1BBL-IL-15 P < 0.0001; Day 6: mRBC-OVA-4-1BBL-IL-15 P < 0.0001, mRBC-OVA-4-1BBL-IL-12 P = 0.0001. d Memory OT-1 number on day 6 in blood. One-way ANOVA compared to mRBC-CTRL; T_scm (stem memory T cells): mRBC-OVA-4-1BBL P = 0.038; T_cm: mRBC-OVA-4-1BBL P = 0.0016, mRBC-OVA-4-1BBL-IL-15 P = 0.025; T_em: mRBC-OVA-4-1BBL-IL-15 P = 0.0001, mRBC-OVA-4-1BBL-IL-15 P < 0.0001. e CD45.1 Pep Boy mice were randomized (~150 mm³; n = 8), treated with naïve OT-1 cells, and dosed with 2.5 × 10⁸ mRBC-CTRL or mRBC-aAPC. f Tumor growth and g survival. Log-rank (Mantel-Cox, one-sided) test of survival curve; mRBC-OVA-4-1BBL vs mRBC-OVA-4-1BBL-IL-12 and PBS vs mRBC-OVA-4-1BBL-IL-12 P < 0.0001. h CD45.1 Pep Boy mice were randomized (~230 mm³; n = 8), treated with naïve OT-1 cells, and dosed with OVA_241-270 peptide with IFA or 1 × 10⁹ mRBC-CTRL, mRBC-4-1BBL-IL-12, or mRBC-OVA-4-1BBL-IL-12. i Tumor growth. j OT-1 number in blood on Day 6. One-way ANOVA compared to mRBC-CTRL; mRBC-OVA-4-1BBL-IL-12 P < 0.0001. k CD45.1 Pep Boy mice were randomized (~175 mm³; n = 5), treated with naïve OT-1 cells, and dosed with 1 × 10⁹ mRBC-CTRL or a dose titration of mRBC-OVA-4-1BBL-IL-12 (1 × 10⁹, 2.5 × 10⁸). l OT-1 numbers in 50 μL blood and per gram of tumor on day 7. One-way ANOVA compared to mRBC-CTRL; blood: 1 × 10⁹ P < 0.0001; tumor: 1 × 10⁹ P = 0.0046. m TCRβ sequencing analyses of OT-1 TCR frequency in blood on day 0 (pre-blood), on day 7 (post-blood), and in tumor on day 7. One-way ANOVA compared to mRBC-CTRL; post blood: 1 × 10⁹ P < 0.0001; tumor: 3 × 10⁸ P = 0.02, 1 × 10⁹ P < 0.0001. n Polyfunctionality (granzyme B⁺IFNγ⁺ %) in the tumor-infiltrating OT-1 cells on day 7. One-way ANOVA compared to mRBC-CTRL; 1 × 10⁹ P = 0.0021. Data are depicted as mean ± s.d. and are representative of four (l blood), two (a, c, d, f, g, l tumor, n), or one (i, j, m) independent experiments. Source data are provided as a Source Data file.