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. 2023 Jul 13;13:11361. doi: 10.1038/s41598-023-37857-9

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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/.

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Histopathological study of RenCa tumors after three different sequential treatment orders with axitinib and immune checkpoint inhibitor (ICI). In vivo subcutaneous tumors were harvested from nude mice after completing treatment according to the schedule shown in this figure. Sections from each tumor tissue were examined by hematoxylin–eosin (H–E) staining and immunohistochemical staining with antibodies against CD3, CD8, and CD11b. (a) Representative findings of histopathological study are shown (scale bar = 100 µm). (b) Quantification of CD8/CD3 ratio in RenCa tumors based on the findings of immunohistochemical staining according to the following treatment group: no treatment, simultaneous administration of axitinib plus anti-mouse programmed death-1 (PD-1) antibody, administration of axitinib followed by anti-mouse PD-1 antibody, and administration of anti-mouse PD-1 antibody followed by axitinib. Each data point represents the mean of eight tumors; bars, standard deviations (SDs). Asterisk, differs from control (p < 0.05) by Student’s t-test. (c) Quantification of CD8/CD11b ratio in RenCa tumors based on the findings of immunohistochemical staining according to the treatment group described in (b). Each data point represents the mean of eight tumors; bars, SDs. Asterisk and double asterisk, differ from control (p < 0.05 and < 0.01, respectively) by Student’s t-test. (d) Quantification of CD8/CD3 ratio in RenCa tumors based on the findings of immunohistochemical staining according to the following treatment groups: no treatment, simultaneous administration of axitinib plus anti-mouse programmed death-ligand 1 (PD-L1) antibody, administration of axitinib followed by anti-mouse PD-L1 antibody, and administration of anti-mouse PD-L1 antibody followed by axitinib. Each data point represents the mean of eight tumors; bars, SDs. Asterisk, differs from control (p < 0.05) by Student’s t-test. (e) Quantification of CD8/CD11b ratio in RenCa tumors based on the findings of immunohistochemical staining according to the treatment group described in (d). Each data point represents the mean of eight tumors; bars, SDs. Asterisk, differs from control (p < 0.01) by Student’s t-test.