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. 2021 Jul 29;14:118. doi: 10.1186/s13045-021-01128-9

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© The Author(s) 2021

Open AccessThis 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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 1 — 7 × 19 CAR-T cells showed enhanced antitumor activity in xenografts. a Schemes of a lentiviral vector of anti-GPC3 CAR, anti-MSLN CAR and anti-CD20 CAR. IL-7 and CCL19 (7 × 19) were linked through the 2A peptide, and truncated CD19 was used to indicate 7 × 19 expression. b The percentage of CAR and 7 × 19 expression on transduced human T cells was analyzed by flow cytometry. c A total of 1 × 10⁶ T cells from each group were cultured in fresh media for 24 h. The supernatant was collected, and the secretion of IL-7 and CCL19 was analyzed by ELISA. The anti-IL-7R antibody and anti-CCR7 antibody were used during culture. Open circle represents the values of individual well. Error bar denotes the S.D. d A total of 1 × 10⁶ anti-GPC3-7 × 19 CAR-T cells or anti-GPC3 CAR-T cells were cultured in the lower chamber of the Transwell plate for 24 h, and then the anti-GPC3 CAR-T cells labeled with CSFE were cultured in the upper chamber for 5 h. The responder T cells that migrated from the upper chamber to the lower chamber were enumerated by flow cytometry. Error bar denoted the S.D. and the results were compared with an unpaired t-test. ^*P < 0.05, ^**P < 0.01, ^***P < 0.001. e A total of 2 × 10⁶ transduced T cells from each group were cultured in fresh media. The cell number was counted every two days. Error bar denotes the S.D. The results were compared with two-way ANOVA. ^*P < 0.05, ^**P < 0.01, ^***P < 0.001. f Tumor volume of HepG2 subcutaneously inoculated mice. Tumor volume = (length × width²)/2. g Tumor weight of HepG2 subcutaneously inoculated mice. h Percentage of T cells in peripheral blood of HepG2 subcutaneously inoculated mice. i Tumor volume of AsPC-1 subcutaneously inoculated mice. j Tumor weight of AsPC-1 subcutaneously inoculated mice. k Percentage of T cells in peripheral blood of AsPC-1 subcutaneously inoculated mice. f, i Error bar denotes the S.D. The results were compared with two-way ANOVA. ^*P < 0.05, ^**P < 0.01, ^***P < 0.001. g, h, j, k The error bar denotes the SD. The results were compared with ordinary one-way ANOVA. ^*P < 0.05, ^**P < 0.01, ^***P < 0.001