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. 2022 Jun 20;72(1):73–85. doi: 10.1007/s00262-022-03226-0

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© The Author(s) 2022

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/.

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Figure. 3 — The absence of BAFF inhibited tumor growth and decreased the numbers of MDSCs and B10 cells. a TC1 cells were transplanted subcutaneously into the right back region of 6- to 8-week-old mice (1*10⁶/100 μl/mouse). The survival time and tumor size were observed every 3 days. b, c Mice were sacrificed on the 28th day of tumor growth, at which point the tumor was removed for measurement and weighing. d The probability of overall survival was significantly different in BAFF WT and BAFF KO groups. Data was analyzed using log-rank (Mantel–Cox) test. e Spleen single-cell suspension was prepared and mononuclear cells were isolated for flow cytometry. The CD19-positive cells were first gated in lymphocytes, and then, CD1d and CD5 were used to gate B10 cells (CD19⁺ CD1d⁺ CD5⁺). Statistical analysis of the frequencies of B10 cells as a percentage of the single-cell suspension. f B10 cells in the spleen were first gated and then the percentage of IL-10⁺ B10 cells were detected. g Tumor single-cell suspension was prepared and mononuclear cells were isolated for flow cytometry. The CD19-positive cells were first gated in lymphocytes, and then, CD1d and CD5 were used to gate B10 cells (CD19⁺ CD1d⁺ CD5⁺). Statistical analysis of the frequencies of B10 cells as a percentage of the single-cell suspension. h B10 cells in the tumor were first gated and then the percentage of IL-10⁺ B10 cells were detected. i–k Single-cell suspension of tumor, spleen and bone marrow were prepared. The CD11b-positive cells were first identified, and then, PMN-MDSCs and M-MDSCs were gated. Statistical analysis of the frequencies of PMN-MDSCs and M-MDSCs as a percentage of the MDSCs population. l The percentage of Annexin V⁺ MDSCs represents the proportion of apoptotic MDSCs. m–p PMN-MDSCs and M-MDSCs were first gated, and then, the percentage of INOS⁺MDSCs were detected. q–r Representative IFN-γ⁺ CD8⁺ T cells flowchart are shown in tumor and spleen tissues from cervical cancer models. Data from at least nine independent experiments were analyzed using Student’s t test and are expressed as the mean ± SD. Symbols represent statistical significance (*p < 0.05)