|Table of Contents|

Exosomes derived from bladder cancer mediate the immunosuppression of cytotoxic T lymphocyte by promoting the amplification of myeloid inhibitory cells

Journal Of Modern Oncology[ISSN:1672-4992/CN:61-1415/R]

Issue:
2022 06
Page:
971-978
Research Field:
Publishing date:

Info

Title:
Exosomes derived from bladder cancer mediate the immunosuppression of cytotoxic T lymphocyte by promoting the amplification of myeloid inhibitory cells
Author(s):
CHEN Jinhua; GU Zuxuan; DENG Li; PAN Weixin
Department of Urology,Hainan Western Central Hospital,Hainan Danzhou 571700，China.
Keywords:
bladder cancer; exosomes; myeloid derived suppressor cells; cytotoxic T lymphocytes; STAT3
PACS:
R737.14
DOI:
10.3969/j.issn.1672-4992.2022.06.005
Abstract:
Objective:To investigate the effect of bladder cancer cell line MB49-derived exosomes (MB49-ExO) on myeloid derived suppressor cells (MDSCs)，and the regulation of the MDSCs on the immune function of MB49 antigen-specific cytotoxic T lymphocytes (CTLs).Methods:MB49-Exo was isolated by the exosome isolation kit,and the exosomes morphology was examined by transmission electron microscope (TEM),and identified by nanoparticle size analyzer (NTA) and Western blot,MB49-Exo tumor-bearing model was established in Balb/c mice.PBS was used as negative control,and MB49 tumor-bearing mice as positive control the espression and typing of MDSCs in bone marrow and spleen and the number of CD8+T.Lymphocytes in spleen were determined by drain cell assay.Primary bone marrow cells of mice were isolated and cultured,and the effects of PBS and MB49-Exo on their differentiation to MDSCs were detected by flow cytometry,and the two groups of MDSCs treated differently were recorded as PBS-MDSCs group and MB49-Exo-MDSCs group.Mice mature dendritic cells (DCs) were induced by MB49-specific antigen and used to prepare bladder cancer antigen specific CTL,which was then stimulated by PBS-MDSCs and MDSCs of the MB49-Exo-MDSCs group.ELISA was used to detect the expression levels of interleukin-1 (IL-2) and interferon-γ (IFN-γ) of CTL under different treatment conditions.CFSE staining combined with flow cytometry was used to detect the proliferation levels of CTL.LDH was used to detect the cytotoxic activity of CTL against MB49.Finally,the phosphorylation levels of signal transduction factor and transcriptional activator 3 (STAT3) in MDSCs of the two groups (p-STAT3/STAT3) were detected by Western blot.Results:MB49-Exo was obtained by successful separation.Compared with the control group,MB49-Exo not only significantly promoted the formation of MDSCs in bone marrow and spleen of tumor-bearing mice (P＜0.001),but also significantly reduced the number of CD8+T lymphocytes in spleen (P＜0.001).In vitro studies,compared with PBS-MDSCs group,MDSCs amplification level was significantly increased in MB49-Exo-MDSCs group (P＜0.001),and the secretion of IL-2 and IFN-γ,cell proliferation ability and cytotoxicity against MB49 of CTL treated by MB49-Exo-MDSCs group were significantly decreased (P＜0.001).However,the phosphorylation level of STAT3 in MB49-Exo-MDSCs group was significantly higher than that in PBS-MDSCs group (P＜0.001).Conclusion:Exosomes from bladder cancer cells could inhibit the immune function of tumor specific CTLs by promoting the formation of MDSCs,thus assisting the immune escape of tumors.The above effects of exosomes may play a role by promoting the activation of STAT3 signaling pathway in MDSCs.

References:

[1] GRAYSON M.Bladder cancer［J］.Nature,2017,551(7679):S33.
[2] SCHNEIDER AK,CHEVALIER MF,DERRE L.The multifaceted immune regulation of bladder cancer［J］.Nat Rev Urol,2019,16(10):613-630.
[3] DYSTHE M,PARIHAR R.Myeloid-derived suppressor cells in the tumor microenvironment［J］.Adv Exp Med Biol,2020,1224:117-140.
[4] 陆源,赵森,杨勇.MDSCs参与肿瘤微环境免疫抑制的研究进展［J］.药学研究,2018,037(012):711-714. LU Y,HAO S,YANG Y.Progress in research on the role of MDSCs in tumor microenvironment immunosuppression［J］.Pharmaceutical Research,2018,037(012):711-714.
[5] PIAO XM,CHA EJ,YUN SJ,et al.Role of exosomal miRNA in bladder cancer:A promising liquid biopsy biomarker［J］.Int J Mol Sci,2021,22(4):e1713.
[6] GUO X,QIU W,WANG J,et al.Glioma exosomes mediate the expansion and function of myeloid-derived suppressor cells through microRNA-29a/Hbp1 and microRNA-92a/Prkar1a pathways［J］.Int J Cancer,2019,144(12):3111-3126.
[7] HUBER V,VALLACCHI V,FLEMING V,et al.Tumor-derived microRNAs induce myeloid suppressor cells and predict immunotherapy resistance in melanoma［J］.J Clin Invest,2018,128(12):5505-5516.
[8] DOMENNIS R,CESSELLI D,TOFFOLETTO B,et al.Systemic T cells immunosuppression of glioma stem cell-derived exosomes is mediated by monocytic myeloid-derived suppressor cells［J］.PLoS One,2017,12(1):e0169932.
[9] 刁建军,宋学东,杨雪,等.膀胱癌MB49细胞exosomes对MDSCs增殖及活性的影响［J］.免疫学杂志,2015,031(002):127-131. DIAO JJ,SONG XD,YANG X,et al.Effects of exosomes on the proliferation and activity of MDSCs in bladder cancer cells［J］.Journal of Immunology,2015,031(002):127-131.
[10] WU T,ZHAO Y,WANG H,et al.mTOR masters monocytic myeloid-derived suppressor cells in mice with allografts or tumors［J］.Sci Rep,2016,6(11):e20250.
[11]谢雪锋.负载凋亡肿瘤细胞抗原的树突状细胞疫苗对小鼠膀胱癌的抑制作用［D］.上海:复旦大学,2014. XIE XF.Inhibitory effect of dendritic cell vaccine loaded with apoptotic tumor cell antigen on mouse bladder cancer［D］.Shanghai:Fudan University,2014.
[12] 余圆圆，方媛，骆莹滨，等.肿瘤来源外泌体抗肿瘤免疫研究进展［J］.肿瘤学杂志,2020,26(3):229-233. YU YY,FANG Y,LUO YB,et al.Research progress in anti-tumor immunity of tumor-derived exosomes［J］.J Oncol,2020,26(3):229-233.
[13] ZHANG L,YU D.Exosomes in cancer development,metastasis,and immunity［J］.Biochim Biophys Acta Rev Cancer,2019,1871(2):455-468.
[14] WHITESIDE TL.Immune modulation of T-cell and NK (natural killer) cell activities by TEXs (tumour-derived exosomes)［J］.Biochem Soc Trans,2013,41(1):245-251.
[15] SHENG IY,DIAZ-MONTERO CM,RAYMAN P,et al.Blood myeloid-derived suppressor cells correlate with neutrophil-to-lymphocyte ratio and overall survival in metastatic urothelial carcinoma［J］.Target Oncol,2020,15(2):211-220.
[16] YANG G,LIU M,LIU Q,et al.Granulocytic myeloid-derived suppressor cells correlate with outcomes undergoing neoadjuvant chemotherapy for bladder cancer［J］.Urol Oncol,2020,38(1):e17-5.e23.
[17] DE SANTO C,SERAFINI P,MARIGO I,et al.Nitroaspirin corrects immune dysfunction in tumor-bearing hosts and promotes tumor eradication by cancer vaccination［J］.Proc Natl Acad Sci USA,2005,102(11):4185-4190.
[18] PAN PY,WANG GX,YIN B,et al.Reversion of immune tolerance in advanced malignancy:modulation of myeloid-derived suppressor cell development by blockade of stem-cell factor function［J］.Blood,2008,111(1):219-228.
[19] 徐浩语，王和西，黎楠，等.Renca细胞来源的外泌体活化髓源抑制性细胞对细胞毒性T细胞的抑制效应［J］.第三军医大学学报,2019,41(10):923-930. XU HY,WANG HX,LI N,et al.Inhibitory effects of renca-derived exosomes activated myeloid suppressor cells on cytotoxic T cells［J］.Journal of the Third Military Medical University,2019,41(10):923-930.
[20] BRUNA H,BOTTCHER M,QORRAJ M,et al.CLL-cell-mediated MDSC induction by exosomal miR-155 transfer is disrupted by vitamin D［J］.Leukemia,2017,31(4):985-988.
[21] DIAO J,YANG X,SONG X,et al.Exosomal Hsp70 mediates immunosuppressive activity of the myeloid-derived suppressor cells via phosphorylation of Stat3［J］.Med Oncol,2015,32(2):e453.
[22] CHALMIN F,LADOIRE S,MIGNOT G,et al.Membrane-associated Hsp72 from tumor-derived exosomes mediates STAT3-dependent immunosuppressive function of mouse and human myeloid-derived suppressor cells［J］.J Clin Invest,2010,120(2):457-471.

Memo

Memo:
海南省卫生健康行业科研项目（编号:20A200290）
Last Update: 1900-01-01