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. 2023 Aug 10;22:131. doi: 10.1186/s12943-023-01830-x

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© The Author(s) 2023

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 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. 6 — Adoptive transfer of bi-specific scFv antibody (BsAb)-secreting CAR-T cells modulate intratumoral cytokines and the proportions of infiltrating immune cells, MDSCs, and M2 TAMs. (A) Representative flow cytometry images and bar summary plots (right) showing the percentage of infiltrating CD8 + CD3 + T cells in tumor-bearing mice. Analysis of cell suspensions extracted from tumors by flow cytometry detecting infiltrating T cells originally gated as CD45 + cells. (B-C) Representative flow cytometry images and bar summary plots (right) showing the percentage of infiltrating MDSCs (B) and M2 TAMs (C) in tumor-bearing mice. Tumor cell suspensions were examined by flow cytometry to detect infiltration of MDSCs marked by CD11b + GR1 + and M2 TAMs marked by F4/80 + CD206+, initially gated as CD45 + cells. (D-E) Representative flow cytometry images and bar summary plots (right) showing the generation of perforin and granzyme B in tumor-infiltrating CD8-positive T cells. The percentages of CD8-positive T cells generating granzyme B (D) and perforin (E) in the TME were analyzed by flow cytometry. (F-I) Intratumoral cytokine evaluation of IL-12, TNF-α, IFN-γ, and IL-15 by ELISA. Data are expressed as means ± SD; * P < 0.05; **P < 0.01, by unpaired Student’s t test. scFv, single-chain fragment variable; CAR-T, chimeric antigen receptor-modified-T; MDSCs, myeloid-derived suppressor cells; TAMs, tumor associated macrophages; CD, cluster of differentiation; TME, tumor microenvironment; IL, interleukin; TNF, tumor necrosis factor; IFN, interferon; ELISA, enzyme-linked immunosorbent assay