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. 2020 Nov 30;11(11):1022. doi: 10.1038/s41419-020-03223-0

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© The Author(s) 2020

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. 3 — A, B Changes of the immune-related cytokines in tumors treated with ZD55-IL-24. A Heat-map of the down-regulated immunosuppressive cytokines in Fig. 2C. B Heat-map of the up-regulated immunostimulatory cytokines in Fig. 2D. C–J Immunohistochemical analysis of immune cell infiltration in tumors, as well as recruitment and activation in spleens. Tumors and spleens resected from B16-bearing C57BL/6 mice receiving PBS or ZD55-IL-24 treatment indicated in Fig. 1A were analyzed 2 days after the last injection by immunohistochemical staining. C Shown are representative images of tumor sections immunostained for CD8 (red, left panel), CD11b and Ly-6G (yellow, middle panel), and NK1.1 (red, right panel). D Quantification of the CD8⁺ cells in C (n = 11). E Quantification of the CD11b⁺Ly-6G⁺Ly-6C^low neutrophils in C (n = 3). f Quantification of the NK1.1⁺ cells in c (n = 6). G Quantification of the CD8⁺ cells in H (n = 9). H Shown are representative images of spleen sections immunostained for CD8 (red, left panel), CD11b and Ly-6G (yellow, middle panel), and NK1.1 (red, right panel). I Quantification of the CD11b⁺Ly-6G⁺Ly-6C^low neutrophils in H (n = 3). j Quantification of NK1.1⁺ cells in h (n = 9). Nuclei is counterstained with DAPI (blue). Data are mean ± SD. Scale bars, 300 µm. Shown is one of three independent experiments. K–M The anti-melanoma efficacy of ZD55-IL-24 in B16-bearing immunocompromised mouse model. BALB/c nude mice were inoculated with B16 tumors and treated with PBS or ZD55-IL-24 as indicated in Fig. 1A. K Shown are tumor growth curves and L photograph of tumors resected from the sacrificed mice at the end of the experiment. M Body weight changes of the mice monitored during the therapy period. P PBS, Z ZD55-IL-24, mean ± SEM is shown. n = 7 mice for PBS-treated group, and n = 10 mice for ZD55-IL-24-treated group. Data represent results from one of two independent experiments.