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. 2023 Jan 9;10:2. doi: 10.21037/sci-2022-030

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2023 Stem Cell Investigation. All rights reserved.

Open Access Statement: This is an Open Access article distributed in accordance with the Creative Commons Attribution-NonCommercial-NoDerivs 4.0 International License (CC BY-NC-ND 4.0), which permits the non-commercial replication and distribution of the article with the strict proviso that no changes or edits are made and the original work is properly cited (including links to both the formal publication through the relevant DOI and the license). See: https://creativecommons.org/licenses/by-nc-nd/4.0.

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Vaccination of ES-exo/GM-CSF suppresses tumor-infiltrating MDSCs and macrophages. Female C57BL/6 mice were vaccinated with PBS or ES-exo or ES-exo/GM-CSF twice (days 0 and 7) and challenged by tail vein inoculation of LLC cells one week later. MDSCs infiltrating into lung tumors were examined 5 weeks after LLC injection. Intratumoral immune cells were identified by CD45, a pan-hematopoietic marker. (A) Dot plots showing the abundance of CD11b⁺ cells in tumor-infiltrating non-T (CD3⁻) and non-B (CD19⁻) immune cells obtained from control, ES-exo and ES-exo/GM-CSF-vaccinated mice. The percentages of the individual subpopulation are shown. (B) The data shown in (A) are summarized with 4 mice in control and ES-exo group, 6 mice in ES-exo/GM-CSF group. Mean ± SD, **, P<0.01; ANOVA with Tukey’s multiple comparison test. (C) Gr-1^high G-MDSCs and Gr-1^low M-MDSCs in CD11b⁺ populations shown in (A) were examined. The percentages of each subpopulation are indicated in the graphs. (D,E) Bar graphs show the summarized data presented in (C) (n=4 in control and ES-exo group, n=6 in ES-exo/GM-CSF group); mean ± SD; **, P<0.01; ANOVA with Tukey’s multiple comparison test. (F) The percentages of F4-80⁺GR-1⁻ macrophages in intratumoral non-T (CD3^–) and non-B (CD19^–) immune cells from control, ES-exo and ES-exo/GM-CSF-vaccinated mice are shown. Numbers in the plots represent the percentages of each subpopulation. (G) Bar graphs showing the average of percentages of F4-80⁺GR-1⁻ macrophages obtained from control, ES-exo and ES-exo/GM-CSF-vaccinated mice (n=4 in control and ES-exo group, n=6 in ES-exo/GM-CSF group); mean ± SD; **, P<0.01; ANOVA with Tukey’s multiple comparison test. ES, embryonic stem; GM-CSF, granulocyte-macrophage colony stimulating factor; n.s. not significant; MDSC, myeloid derived suppressor cell; PBS, phosphate buffered saline; LLC, Lewis lung carcinoma; SD, standard deviation; ANOVA, analysis of variance; G-MDSC, granulocytic MDSC; M-MDSC, monocytic MDSC.