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. 2019 Aug 23;9:12308. doi: 10.1038/s41598-019-48711-2

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© The Author(s) 2019

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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Sequential addition of fresh CTL cohorts results in better control of tumor growth in vivo. C57BL/6 mice were injected subcutaneously with 2 × 10⁶ OVA⁺ B16 tumor cells. On day 5, mice were adoptively transferred with 6 × 10⁶ activated OT-I CD8⁺ T cells either in 1 injection or in 3 injections of 2 × 10⁶ each with a time interval of 4 hours. A third group served as control with no CTL injection. Tumor growth was monitored over time. Data are from three independent experiments performed on 3 mice per group and are represented as mean +/− SD. Data was analyzed by two-way repeated measures ANOVA using the GraphPad Prism software. ***P < 0.0001.