华西口腔医学杂志

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肿瘤exosome诱导特异性T杀伤细胞的研究

王东关;孙善珍;王振光;王霞   

  1. 山东大学口腔医院病理科,山东济南250012
  • 收稿日期:2006-04-25 修回日期:2006-04-25 出版日期:2006-04-20 发布日期:2006-04-20
  • 通讯作者: 孙善珍, Tel:0531-88382057
  • 作者简介:王东关(1970-),男,山东人,主治医师,博士研究生
  • 基金资助:

    高等学校博士学科点专项科研基金资助项目(20030422041)

Study of Induction of Tumor Specific Cytotoxic T Lymphocyte by Using Tumor-derived Exosome

WANG Dong-guan,SUN Shan-zhen,WANG Zhen-guang,WANG Xia   

  1. Dept. of Pathology, School of Stomatology, Shandong University, Jinan 250012, China
  • Received:2006-04-25 Revised:2006-04-25 Online:2006-04-20 Published:2006-04-20

摘要:

目的检测癌细胞来源的exosome在体外刺激细胞毒性T淋巴细胞(CTL)特异性杀伤癌细胞的能力.方法分离纯化舌鳞癌Tca8113细胞,培养上清液中的exosome,制备癌细胞的冻融抗原(FTA),并把两者负载到从血液分离和培养的树突状细胞(DC)上,测定其诱导的CTL对Tca8113的杀伤活性,以SPCA-1人肺腺癌细胞系和95-D人巨细胞癌为对照组.结果体外FTA和exosome冲击致敏的DC能显著刺激T淋巴细胞增殖,其诱导的CTL对细胞系Tca8113具有显著的杀伤作用,在效靶比为20:1时12 h平均分别为45.19%+4.57%和43.60%+5.66%(P<0.01);exosome冲击致敏的DC诱导的CTL对SPCA-1人肺腺癌也有明显的杀伤作用(P<0.05),但FTA冲击致敏的DC诱导的CTL却无此功能.结论肿瘤细胞培养上清液中分离出的exosome,负载到DC上后,可以活化T细胞,使之成为抗原特异性的CTL,具有特异性的杀伤肿瘤细胞的功能,为口腔癌的免疫治疗开拓了新的途径.exosome特异的CTL也可杀死其他肿瘤细胞,说明肿瘤来源的exosome是一种可引起肿瘤消退的共同抗原.

关键词: Tca8113细胞, 细胞毒性T淋巴细胞, 树突状细胞

Abstract:

Objective To investigate whether the exosomes derived from Tca8113 could induce production of tumor-specific  T cells when pulsed onto dendritic cells. Methods  Tca8113 cell was cultured with RPIMI640. isolated and purified the tumor-derived exosomes from the culture supernatants by ultrafiltration with Millipore centrifual filter devices; frozen and thawed Tca8113 cells to get frozen tumor antigens(FTA). The exosomes and FT A was pulsed onto DC generated from normal human peripheral blood mononuclear cell (PBMC) in vitro. The DC pulsed with FAP or exosomes cocultured with the peripheral blood lymphocytes to transform T cell into specific CTL To observe the killing and  wounding activity of CTL, the CTL and Tca8113 cells were mixed at a ratio of 20 to 1, SPCA-l cells and 95-D cells was evaluated as control group. Results The CTL induced by DC pulsed with FAP or exosomes had significant activity killing Tca8113 CP

Key words: Tca8113cell, cytotoxicTlymphocyte, dendriticcell