The Journal of Practical Medicine >
Effect of culture time on immune⁃related membrane proteins of mouse dendritic cells and their exosomes
Received date: 2023-10-19
Online published: 2024-04-08
Objective This study aims to investigate the impact of cultivation time on dendritic cells (DCs) and their derived exosomes' expression of immune?related membrane proteins (CD80, MHC?Ⅰ, MHC?Ⅱ) and provides experimental evidence for future research. Methods Mouse bone marrow cells were induced to differentiate into DCs using GM?CSF and IL?4, followed by maturation stimulation withTNF?α. Exosomes were extracted using ultracentrifugation.Western blot and Amnis image flow cytometry were used to identify exosomes derived from mouse DCs. Amnis image flow cytometry was used to detect the expression of immune?related proteins CD80, CD11c, MHC?Ⅰ, and MHC?Ⅱ in mouse DCs and their exosomes. Results After 5 days of in vitro cultivation, more than 50% of dendritic cells expressed CD80, CD11c, MHC?Ⅰ, and MHC?Ⅱ, reaching the highest level on day 13. The positivity rates were as follows: CD80 (97.29 ± 0.63) %, CD11c (92.31 ± 1.18) %, MHC?Ⅰ(97.91 ± 0.49) %, and MHC?Ⅱ(97.91 ± 0.49) %. The differences were statistically significant (P < 0.001). The expression gradually decreased after day 13, but approximately 80% of DC cells still expressed MHC?Ⅰ and MHC?Ⅱ immune molecules on day 30. The expression levels of CD80, CD11c, and MHC?Ⅱ on the exosome membrane were highest on day 5 and then decreased overall with prolonged cultivation time, except for MHC?Ⅰ molecules. The differences were statistically significant (P < 0.01). Conclusions In vitro?cultured mouse dendritic cells express high levels of immune?related membrane proteins and can be stably maintained for a long time under suitable culture conditions. The secreted exosomes also carry abundant immune?related membrane proteins, but no significant correlation was found between the immune?related proteins on the dendritic cell surface and the exosome membrane surface.
Key words: mouse dendritic cells; exosomes; immune membrane proteins
Li XIAO , Shumin LUO , Fang XU , Pengpeng LU , Enhong XING , Weihua. LI . Effect of culture time on immune⁃related membrane proteins of mouse dendritic cells and their exosomes[J]. The Journal of Practical Medicine, 2024 , 40(7) : 941 -947 . DOI: 10.3969/j.issn.1006-5725.2024.07.011
| 1 | WCULEK S K, CUETO F J, MUJAL A M,et al. Dendritic cells in cancer immunology and immunotherapy[J]. Nat Rev Immunol, 2020,20(1):7-24. doi:10.1038/s41577-019-0210-z |
| 2 | HILLGAN K L, RONCHESE F. Antigen presentation by dendritic cells and their instruction of CD4+ T helper cell responses[J]. Cell Mol Immunol, 2020,17(6):587-599. doi:10.1038/s41423-020-0465-0 |
| 3 | GALATI D, ZANOTTA S. Dendritic Cell and Cancer Therapy[J]. Int J Mol Sci, 2023,24(4):4253. doi:10.3390/ijms24044253 |
| 4 | LI X, LI C, ZHANG L, et al.The significance of exosomes in the development and treatment of hepatocellular carcinoma[J]. Mol Cancer, 2020,19(1):1. doi:10.1186/s12943-019-1085-0 |
| 5 | GHORBANINEZHAD F, ALEMOHAMMAD H, NAJAFZADEH B, et al. Dendritic cell-derived exosomes: A new horizon in personalized cancer immunotherapy[J]. Cancer Lett, 2023,562:216168. doi:10.1016/j.canlet.2023.216168 |
| 6 | 张芸蔓,黄功华,王妍妍. 树突状细胞在过敏性哮喘发病中的作用及机制研究新进展[J]. 实用医学杂志, 2023,39(17):2280-2284. doi:10.3969/j.issn.1006-5725.2023.17.023 |
| 7 | 裴天楚,黄隆,吴国惠,等. 树突状细胞疫苗治疗口腔鳞状细胞癌的研究进展[J]. 实用医学杂志, 2021,37(4):419-422. doi:10.3969/j.issn.1006-5725.2021.04.001 |
| 8 | TIBERIO L, DEL PRETE A, SCHIOPPA T, et al.Chemokine and chemotactic signals in dendritic cell migration[J]. Cell Mol Immunol, 2018,15(4):346-352. doi:10.1038/s41423-018-0005-3 |
| 9 | WANG Y, XIANG Y, XIN V W, et al. Dendritic cell biology and its role in tumor immunotherapy[J]. J Hematol Oncol, 2020,13(1):107. doi:10.1186/s13045-020-00939-6 |
| 10 | LI J, LI J, PENG Y, et al. Dendritic cell derived exosomes loaded neoantigens for personalized cancer immunotherapies[J]. J Control Release, 2023,353:423-433. doi:10.1016/j.jconrel.2022.11.053 |
| 11 | LUO S, CHEN J, XU F, et al. Dendritic Cell-Derived Exosomes in Cancer Immunotherapy[J]. Pharmaceutics, 2023,15(8):2070. doi:10.3390/pharmaceutics15082070 |
| 12 | XIA J, MIAO Y, WANG X, et al. Recent progress of dendritic cell-derived exosomes (Dex) as an anti-cancer nanovaccine[J]. Biomed Pharmacother, 2022,152:113250. doi:10.1016/j.biopha.2022.113250 |
| 13 | NIKFARJAM S, REZAIE J, KASHANCHI F, et al. Dexosomes as a cell-free vaccine for cancer immunotherapy[J]. J Exp Clin Cancer Res, 2020,39(1):258. doi:10.1186/s13046-020-01781-x |
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