{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,18]],"date-time":"2026-08-18T04:47:40Z","timestamp":1787028460842,"version":"build-2736575974"},"reference-count":64,"publisher":"MDPI AG","issue":"20","license":[{"start":{"date-parts":[[2021,10,13]],"date-time":"2021-10-13T00:00:00Z","timestamp":1634083200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100012166","name":"National Key Research and Development Program of China","doi-asserted-by":"publisher","award":["2020YFA0909003"],"award-info":[{"award-number":["2020YFA0909003"]}],"id":[{"id":"10.13039\/501100012166","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["82072085"],"award-info":[{"award-number":["82072085"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100003399","name":"Science and Technology Commission of Shanghai Municipality","doi-asserted-by":"publisher","award":["19DZ 2280300"],"award-info":[{"award-number":["19DZ 2280300"]}],"id":[{"id":"10.13039\/501100003399","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Effective cancer therapies should reshape immunosuppression and trigger antitumor immunity. Previously, we developed a novel cryo-thermal therapy through applying local rapid cooling followed by rapid heating of tumor tissue. It could not only ablate local tumors, but also, subsequently, induce systemic long-term antitumor immunity. Hyperthermia can induce the release of extracellular vesicles (EVs) to stimulate antitumor immunity. We examine whether EVs are released after cryo-thermal therapy and whether they could improve the efficacy of cryo-thermal therapy in the 4T1 model. In this study, serum extracellular vesicles (sEVs) are isolated and characterized 3 h after cryo-thermal therapy of subcutaneous tumors. sEV phagocytosis is observed in vitro and in vivo by using laser confocal microscopy and flow cytometry. After cryo-thermal therapy, sEVs are administered to mice via the tail vein, and changes in immune cells are investigated by using flow cytometry. After cryo-thermal therapy, a large number of sEVs are released to the periphery carrying danger signals and tumor antigens, and these sEVs could be phagocytosed by peripheral blood monocytes and differentiated macrophages. After cryo-thermal therapy, supplementation with sEVs released after treatment promotes the differentiation of myeloid-derived suppressor cells (MDSCs), monocytes into macrophages and CD4+ T cells into the Th1 subtype, as well as prolonging the long-term survival of the 4T1 subcutaneous tumor-bearing mice. sEVs released after cryo-thermal tumor treatment could clinically serve as an adjuvant in subsequent cryo-thermal therapy to improve the therapeutic effects on malignant tumors.<\/jats:p>","DOI":"10.3390\/ijms222011021","type":"journal-article","created":{"date-parts":[[2021,10,13]],"date-time":"2021-10-13T21:36:15Z","timestamp":1634160975000},"page":"11021","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":7,"title":["Supplementation with Serum-Derived Extracellular Vesicles Reinforces Antitumor Immunity Induced by Cryo-Thermal Therapy"],"prefix":"10.3390","volume":"22","author":[{"given":"Yinuo","family":"Cen","sequence":"first","affiliation":[{"name":"School of Biomedical Engineering, Shanghai Jiao Tong University, Shanghai 200030, China"},{"name":"School of Biomedical Engineering, Med-X Research Institute, Shanghai Jiao Tong University, Shanghai 200030, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yue","family":"Lou","sequence":"additional","affiliation":[{"name":"School of Biomedical Engineering, Shanghai Jiao Tong University, Shanghai 200030, China"},{"name":"School of Biomedical Engineering, Med-X Research Institute, Shanghai Jiao Tong University, Shanghai 200030, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Junjun","family":"Wang","sequence":"additional","affiliation":[{"name":"School of Biomedical Engineering, Shanghai Jiao Tong University, Shanghai 200030, China"},{"name":"School of Biomedical Engineering, Med-X Research Institute, Shanghai Jiao Tong University, Shanghai 200030, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shicheng","family":"Wang","sequence":"additional","affiliation":[{"name":"School of Biomedical Engineering, Shanghai Jiao Tong University, Shanghai 200030, China"},{"name":"School of Biomedical Engineering, Med-X Research Institute, Shanghai Jiao Tong University, Shanghai 200030, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Peng","family":"Peng","sequence":"additional","affiliation":[{"name":"School of Biomedical Engineering, Shanghai Jiao Tong University, Shanghai 200030, China"},{"name":"School of Biomedical Engineering, Med-X Research Institute, Shanghai Jiao Tong University, Shanghai 200030, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Aili","family":"Zhang","sequence":"additional","affiliation":[{"name":"School of Biomedical Engineering, Shanghai Jiao Tong University, Shanghai 200030, China"},{"name":"School of Biomedical Engineering, Med-X Research Institute, Shanghai Jiao Tong University, Shanghai 200030, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ping","family":"Liu","sequence":"additional","affiliation":[{"name":"School of Biomedical Engineering, Shanghai Jiao Tong University, Shanghai 200030, China"},{"name":"School of Biomedical Engineering, Med-X Research Institute, Shanghai Jiao Tong University, Shanghai 200030, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2021,10,13]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"667","DOI":"10.1038\/s41568-019-0209-6","article-title":"The pro-tumorigenic host response to cancer therapies","volume":"19","author":"Shaked","year":"2019","journal-title":"Nat. Rev. Cancer"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"611","DOI":"10.1038\/nrclinonc.2016.57","article-title":"Balancing efficacy of and host immune responses to cancer therapy: The yin and yang effects","volume":"13","author":"Shaked","year":"2016","journal-title":"Nat. Rev. Clin. Oncol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"109","DOI":"10.1038\/cr.2016.151","article-title":"Regulatory T cells in cancer immunotherapy","volume":"27","author":"Tanaka","year":"2017","journal-title":"Cell Res."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"16","DOI":"10.1016\/j.critrevonc.2013.07.008","article-title":"Surgery, wound healing, and metastasis: Recent insights and clinical implications","volume":"89","author":"Ceelen","year":"2014","journal-title":"Crit. Rev. Oncol. Hematol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"565","DOI":"10.1016\/j.iac.2014.04.003","article-title":"Adverse events to nontargeted and targeted chemotherapeutic agents: Emphasis on hypersensitivity responses","volume":"34","author":"Baldo","year":"2014","journal-title":"Immunol. Allergy Clin. N. Am."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"25","DOI":"10.1038\/s41577-019-0218-4","article-title":"Tumour-intrinsic resistance to immune checkpoint blockade","volume":"20","author":"Kalbasi","year":"2020","journal-title":"Nat. Rev. Immunol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"807","DOI":"10.1038\/s41423-020-0488-6","article-title":"The history and advances in cancer immunotherapy: Understanding the characteristics of tumor-infiltrating immune cells and their therapeutic implications","volume":"17","author":"Zhang","year":"2020","journal-title":"Cell Mol. Immunol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"443","DOI":"10.1016\/j.ccell.2020.03.017","article-title":"Acquired Resistance to Immune Checkpoint Inhibitors","volume":"37","author":"Schoenfeld","year":"2020","journal-title":"Cancer Cell"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"199","DOI":"10.1038\/nrc3672","article-title":"Thermal ablation of tumours: Biological mechanisms and advances in therapy","volume":"14","author":"Chu","year":"2014","journal-title":"Nat. Rev. Cancer"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"899","DOI":"10.1038\/s41467-019-08782-1","article-title":"Irreversible electroporation reverses resistance to immune checkpoint blockade in pancreatic cancer","volume":"10","author":"Zhao","year":"2019","journal-title":"Nat. Commun."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"528","DOI":"10.3109\/02656736.2012.677933","article-title":"Old and new facts about hyperthermia-induced modulations of the immune system","volume":"28","author":"Frey","year":"2012","journal-title":"Int. J. Hyperth."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"1174","DOI":"10.2174\/156652412803306666","article-title":"Dual role of heat shock proteins (HSPs) in anti-tumor immunity","volume":"12","author":"Multhoff","year":"2012","journal-title":"Curr. Mol. Med."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"4","DOI":"10.3109\/02656736.2015.1091093","article-title":"The future of biology in driving the field of hyperthermia","volume":"32","author":"Dewhirst","year":"2016","journal-title":"Int. J. Hyperth."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"5478","DOI":"10.1016\/j.ijheatmasstransfer.2008.04.027","article-title":"Evaluation of alternate cooling and heating for tumor treatment","volume":"51","author":"Sun","year":"2008","journal-title":"Int. J. Heat Mass Transf."},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Sun, J.Q., Xu, C.C., Wei, G.H., Sun, X.G., Liu, P., Zhang, A.L., and Xu, L.X. (2009, January 7\u201312). Tumor Treatment System with Alternate Cooling and Heating\u2014Preliminary Results in an Animal Model. Proceedings of the World Congress on Medical Physics and Biomedical Engineering, Munich, Germany.","DOI":"10.1007\/978-3-642-03906-5_93"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"534","DOI":"10.1080\/10407782.2013.742809","article-title":"Numerical Simulation of a New Probe for the Alternate Cooling and Heating of a Subcutaneous Mouse Tumor Model","volume":"63","author":"Cai","year":"2013","journal-title":"Numer. Heat Transf. Part A Appl."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"27136","DOI":"10.1038\/srep27136","article-title":"Cryo-thermal therapy elicits potent anti-tumor immunity by inducing extracellular Hsp70-dependent MDSC differentiation","volume":"6","author":"Zhu","year":"2016","journal-title":"Sci. Rep."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"843","DOI":"10.1080\/02656736.2020.1788173","article-title":"Tumor-related HSP70 released after cryo-thermal therapy targeted innate immune initiation in the antitumor immune response","volume":"37","author":"Zhu","year":"2020","journal-title":"Int. J. Hyperth."},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Peng, P., Hu, H., Liu, P., and Xu, L.X. (2020). Neoantigen-specific CD4 T-cell response is critical for the therapeutic efficacy of cryo-thermal therapy. J. Immunother. Cancer, 8.","DOI":"10.1136\/jitc-2019-000421"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"531","DOI":"10.3109\/02656736.2014.968640","article-title":"Local tumour hyperthermia as immunotherapy for metastatic cancer","volume":"30","author":"Fiering","year":"2014","journal-title":"Int. J. Hyperth."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"1598","DOI":"10.1002\/eji.200535501","article-title":"Efficient induction of antitumor T cell immunity by exosomes derived from heat-shocked lymphoma cells","volume":"36","author":"Chen","year":"2006","journal-title":"Eur. J. Immunol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"2219","DOI":"10.4049\/jimmunol.1002991","article-title":"Chemokine-containing exosomes are released from heat-stressed tumor cells via lipid raft-dependent pathway and act as efficient tumor vaccine","volume":"186","author":"Chen","year":"2011","journal-title":"J. Immunol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"373","DOI":"10.1083\/jcb.201211138","article-title":"Extracellular vesicles: Exosomes, microvesicles, and friends","volume":"200","author":"Raposo","year":"2013","journal-title":"J. Cell Biol."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"195","DOI":"10.1038\/nri3622","article-title":"Regulation of immune responses by extracellular vesicles","volume":"14","author":"Robbins","year":"2014","journal-title":"Nat. Rev. Immunol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"604","DOI":"10.3109\/02656736.2011.564598","article-title":"Induction of a tumour-specific CTL response by exosomes isolated from heat-treated malignant ascites of gastric cancer patients","volume":"27","author":"Zhong","year":"2011","journal-title":"Int. J. Hyperth."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"132","DOI":"10.1111\/imm.12874","article-title":"Exosomes from heat-stressed tumour cells inhibit tumour growth by converting regulatory T cells to Th17 cells via IL-6","volume":"154","author":"Guo","year":"2018","journal-title":"Immunology"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"216","DOI":"10.1038\/s41419-019-1459-7","article-title":"Cryo-thermal therapy induces macrophage polarization for durable anti-tumor immunity","volume":"10","author":"He","year":"2019","journal-title":"Cell Death Dis."},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Pulaski, B.A., and Ostrand-Rosenberg, S. (2001). Mouse 4T1 breast tumor model. Curr. Protoc. Immunol.","DOI":"10.1002\/0471142735.im2002s39"},{"key":"ref_29","doi-asserted-by":"crossref","unstructured":"Mulcahy, L.A., Pink, R.C., and Carter, D.R. (2014). Routes and mechanisms of extracellular vesicle uptake. J. Extracell. Vesicles, 3.","DOI":"10.3402\/jev.v3.24641"},{"key":"ref_30","doi-asserted-by":"crossref","unstructured":"Th\u00e9ry, C., Amigorena, S., Raposo, G., and Clayton, A. (2006). Isolation and characterization of exosomes from cell culture supernatants and biological fluids. Curr. Protoc. Cell Biol.","DOI":"10.1002\/0471143030.cb0322s30"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"399","DOI":"10.1038\/nrclinonc.2016.217","article-title":"Tumour-associated macrophages as treatment targets in oncology","volume":"14","author":"Mantovani","year":"2017","journal-title":"Nat. Rev. Clin. Oncol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"162","DOI":"10.1038\/nri2506","article-title":"Myeloid-derived suppressor cells as regulators of the immune system","volume":"9","author":"Gabrilovich","year":"2009","journal-title":"Nat. Rev. Immunol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"394","DOI":"10.1038\/cr.2012.178","article-title":"A novel role of hematopoietic CCL5 in promoting triple-negative mammary tumor progression by regulating generation of myeloid-derived suppressor cells","volume":"23","author":"Zhang","year":"2013","journal-title":"Cell Res."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"349","DOI":"10.1038\/nri.2017.28","article-title":"Monocyte differentiation and antigen-presenting functions","volume":"17","author":"Jakubzick","year":"2017","journal-title":"Nat. Rev. Immunol."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"579","DOI":"10.1146\/annurev-immunol-042617-053411","article-title":"CD4 Helper and CD8 Cytotoxic T Cell Differentiation","volume":"36","author":"Taniuchi","year":"2018","journal-title":"Annu. Rev. Immunol."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"14","DOI":"10.3389\/fimmu.2018.00014","article-title":"Recent Advances in Targeting CD8 T-Cell Immunity for More Effective Cancer Immunotherapy","volume":"9","author":"Durgeau","year":"2018","journal-title":"Front. Immunol."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"1806444","DOI":"10.1080\/20013078.2020.1806444","article-title":"A combined \u201ceat me\/don\u2019t eat me\u201d strategy based on extracellular vesicles for anticancer nanomedicine","volume":"9","author":"Belhadj","year":"2020","journal-title":"J. Extracell. Vesicles"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"612","DOI":"10.1126\/science.1175202","article-title":"Identification of splenic reservoir monocytes and their deployment to inflammatory sites","volume":"325","author":"Swirski","year":"2009","journal-title":"Science"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"15076","DOI":"10.1038\/s41598-018-33440-9","article-title":"CXCR1 differentiates F4\/80 monocytes into pro-inflammatory F4\/80 macrophages in the liver","volume":"8","author":"Lee","year":"2018","journal-title":"Sci. Rep."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"123","DOI":"10.1016\/j.yjmcc.2015.12.028","article-title":"Exosomes derived from dendritic cells improve cardiac function via activation of CD4(+) T lymphocytes after myocardial infarction","volume":"91","author":"Liu","year":"2016","journal-title":"J. Mol. Cell Cardiol."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"3538","DOI":"10.1016\/j.vaccine.2011.02.095","article-title":"GP120-specific exosome-targeted T cell-based vaccine capable of stimulating DC- and CD4(+) T-independent CTL responses","volume":"29","author":"Nanjundappa","year":"2011","journal-title":"Vaccine"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"266","DOI":"10.1016\/j.molimm.2019.06.006","article-title":"TLR1\/TLR2 signaling blocks the suppression of monocytic myeloid-derived suppressor cell by promoting its differentiation into M1-type macrophage","volume":"112","author":"Deng","year":"2019","journal-title":"Mol. Immunol."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"578","DOI":"10.1038\/mi.2010.41","article-title":"TLR4\/MyD88-induced CD11b+Gr-1 int F4\/80+ non-migratory myeloid cells suppress Th2 effector function in the lung","volume":"3","author":"Arora","year":"2010","journal-title":"Mucosal. Immunol."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"e1230578","DOI":"10.1080\/2162402X.2016.1230578","article-title":"TLR7-based cancer immunotherapy decreases intratumoral myeloid-derived suppressor cells and blocks their immunosuppressive function","volume":"5","author":"Spinetti","year":"2016","journal-title":"Oncoimmunology"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"597","DOI":"10.1016\/j.immuni.2020.07.001","article-title":"High-Dimensional T Helper Cell Profiling Reveals a Broad Diversity of Stably Committed Effector States and Uncovers Interlineage Relationships","volume":"53","author":"Tortola","year":"2020","journal-title":"Immunity"},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"91","DOI":"10.1158\/2326-6066.CIR-13-0216","article-title":"CD4 T-cell subsets and tumor immunity: The helpful and the not-so-helpful","volume":"2","author":"Kim","year":"2014","journal-title":"Cancer Immunol. Res."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"651","DOI":"10.1084\/jem.20091921","article-title":"Naive tumor-specific CD4(+) T cells differentiated in vivo eradicate established melanoma","volume":"207","author":"Xie","year":"2010","journal-title":"J. Exp. Med."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"637","DOI":"10.1084\/jem.20091918","article-title":"Tumor-reactive CD4(+) T cells develop cytotoxic activity and eradicate large established melanoma after transfer into lymphopenic hosts","volume":"207","author":"Quezada","year":"2010","journal-title":"J. Exp. Med."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"281","DOI":"10.1038\/ni.2523","article-title":"Transcriptional reprogramming of mature CD4\u207a helper T cells generates distinct MHC class II-restricted cytotoxic T lymphocytes","volume":"14","author":"Mucida","year":"2013","journal-title":"Nat. Immunol."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"17","DOI":"10.1016\/j.canlet.2016.12.039","article-title":"MHC class II restricted neoantigen: A promising target in tumor immunotherapy","volume":"392","author":"Sun","year":"2017","journal-title":"Cancer Lett."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"219","DOI":"10.3389\/fimmu.2019.00219","article-title":"Cognate Interaction with CD4 T Cells Instructs Tumor-Associated Macrophages to Acquire M1-Like Phenotype","volume":"10","author":"Eisel","year":"2019","journal-title":"Front. Immunol."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"480","DOI":"10.1038\/31002","article-title":"T-cell help for cytotoxic T lymphocytes is mediated by CD40\u2013CD40L interactions","volume":"393","author":"Schoenberger","year":"1998","journal-title":"Nature"},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"988","DOI":"10.1002\/eji.201445036","article-title":"B cells expressing IFN-\u03b3 suppress Treg-cell differentiation and promote autoimmune experimental arthritis","volume":"45","author":"Olalekan","year":"2015","journal-title":"Eur. J. Immunol."},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"e2703","DOI":"10.1038\/cddis.2017.125","article-title":"The cryo-thermal therapy eradicated melanoma in mice by eliciting CD4 T-cell-mediated antitumor memory immune response","volume":"8","author":"He","year":"2017","journal-title":"Cell Death Dis."},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"14852","DOI":"10.1073\/pnas.1403271111","article-title":"Antigen affinity and antigen dose exert distinct influences on CD4 T-cell differentiation","volume":"111","author":"Keck","year":"2014","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"64","DOI":"10.1158\/2326-6066.CIR-13-0030","article-title":"Myeloid-derived suppressor cells attenuate TH1 development through IL-6 production to promote tumor progression","volume":"1","author":"Tsukamoto","year":"2013","journal-title":"Cancer Immunol. Res."},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"e13081","DOI":"10.1111\/aji.13081","article-title":"Altered crosstalk of estradiol and progesterone with Myeloid-derived suppressor cells and Th1\/Th2 cytokines in early miscarriage is associated with early breakdown of maternal-fetal tolerance","volume":"81","author":"Verma","year":"2019","journal-title":"Am. J. Reprod. Immunol."},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"513","DOI":"10.1158\/0008-5472.CAN-15-1737","article-title":"A Breakthrough: Macrophage-Directed Cancer Immunotherapy","volume":"76","author":"Mills","year":"2016","journal-title":"Cancer Res."},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"89","DOI":"10.1111\/imr.12035","article-title":"The Th17 family: Flexibility follows function","volume":"252","author":"Basu","year":"2013","journal-title":"Immunol. Rev."},{"key":"ref_60","doi-asserted-by":"crossref","first-page":"e12128","DOI":"10.1002\/jev2.12128","article-title":"Molecular Evaluation of Five Different Isolation Methods for Extracellular Vesicles Reveals Different Clinical Applicability and Subcellular Origin","volume":"10","author":"Veerman","year":"2021","journal-title":"J. Extracell. Vesicles"},{"key":"ref_61","doi-asserted-by":"crossref","first-page":"234","DOI":"10.1016\/j.aca.2020.06.071","article-title":"Separation and Characterization of Extracellular Vesicles from Human Plasma by Asymmetrical Flow Field-Flow Fractionation","volume":"1127","author":"Wu","year":"2020","journal-title":"Anal. Chim. Acta"},{"key":"ref_62","doi-asserted-by":"crossref","first-page":"2873","DOI":"10.1007\/s00018-018-2773-4","article-title":"Detailed Analysis of the Plasma Extracellular Vesicle Proteome after Separation from Lipoproteins","volume":"75","author":"Karimi","year":"2018","journal-title":"Cell Mol. Life Sci."},{"key":"ref_63","doi-asserted-by":"crossref","first-page":"697","DOI":"10.1038\/s41568-020-00299-w","article-title":"The evolving translational potential of small extracellular vesicles in cancer","volume":"20","author":"Lobb","year":"2020","journal-title":"Nat. Rev. Cancer"},{"key":"ref_64","doi-asserted-by":"crossref","first-page":"1535750","DOI":"10.1080\/20013078.2018.1535750","article-title":"Minimal information for studies of extracellular vesicles 2018 (MISEV2018): A position statement of the International Society for Extracellular Vesicles and update of the MISEV2014 guidelines","volume":"7","author":"Witwer","year":"2018","journal-title":"J. Extracell. Vesicles"}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/22\/20\/11021\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T07:12:24Z","timestamp":1760166744000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/22\/20\/11021"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,10,13]]},"references-count":64,"journal-issue":{"issue":"20","published-online":{"date-parts":[[2021,10]]}},"alternative-id":["ijms222011021"],"URL":"https:\/\/doi.org\/10.3390\/ijms222011021","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,10,13]]}}}