Du Ying, Chen Shao-qian, Huang Yu-min, Dong Zi-ming
Department of Microbiology and Immunology, College of Basic Medical Sciences, Zhengzhou University, Zhengzhou 450052, China.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2006 Jan;22(1):33-6.
To investigate the properties of exosomes secreted by dendritic cells(DCs) derived from human umbilical cord blood mononuclear cells(HUBMCs) and its enhancement of CTL cytotoxicity.
The HUBMCs were separated from human umbilical cord blood(HUCB). The generation of DCs were induced for 12 days in medium containing SCF, IL-4 and GM-CSF. Tumor antigen extracted by repeated freeze-thawing were added to culture medium and cultured successively for 2 days. The DCs were collected and exosomes were separated from culture supernatant through centrifugation at 100,000 g. The exosomes-associated properties were analyzed by FCM, SDS-PAGE, transmission electron microscope(TEM) and MTT colorimetry.
After being induced for 12 days with SCF, IL-4 and GM-CSF the number of DCs had a 10-fold increase and displayed typically morphological characteristics of DCs. FCM analysis showed that the DCs loaded with tumor antigen expressed highly MHC-I, MHC-II, CD40, CD80, CD86, CD11c and CD54. Exosomes could be separated from culture supernatant of DCs by ultracentrifugation. Mixed lymphocyte reation(MLR) and cytotoxicity assay showed that exosomes secreted by DCs could effectively stimulate T cell proliferation and enhance their cytotoxicity.
A large number of DCs are obtained from UBMCs and the exosomes are secreted from them in vitro. The exosomes can present tumor antigen to T cells and enhance the cytotoxicity of CTLs.
研究人脐带血单个核细胞（HUBMCs）来源的树突状细胞（DCs）分泌的外泌体的特性及其对CTL细胞毒性的增强作用。
从人脐带血（HUCB）中分离出HUBMCs。在含有SCF、IL-4和GM-CSF的培养基中诱导DCs生成12天。将反复冻融提取的肿瘤抗原加入培养基中并连续培养2天。收集DCs，通过100,000 g离心从培养上清液中分离出外泌体。通过流式细胞术（FCM）、十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）、透射电子显微镜（TEM）和MTT比色法分析外泌体相关特性。
用SCF、IL-4和GM-CSF诱导12天后，DCs数量增加了10倍，并呈现出典型的DCs形态特征。FCM分析显示，负载肿瘤抗原的DCs高表达MHC-I、MHC-II、CD40、CD80、CD86、CD11c和CD54。外泌体可通过超速离心从DCs的培养上清液中分离出来。混合淋巴细胞反应（MLR）和细胞毒性测定表明，DCs分泌的外泌体可有效刺激T细胞增殖并增强其细胞毒性。
从脐带血单个核细胞中获得了大量DCs，并在体外从中分泌出外泌体。外泌体可将肿瘤抗原呈递给T细胞并增强CTL的细胞毒性。
