癌变·畸变·突变 ›› 2007, Vol. 19 ›› Issue (4): 290-293.doi: 10.3969/j.issn.1004-616x.2007.04.008

• 论著 • 上一篇    下一篇

Anti_CD20_scFv_IgGFc基因修饰T细胞诱导Raji细胞凋亡的研究

潘智勇 谭映霞 俞 康 吴建波 章圣辉 韩义香   

  1. 温州医学院生物学实验教学中心,浙江 温州
  • 收稿日期:2006-10-08 修回日期:2007-03-03 出版日期:2007-07-30 发布日期:2007-07-30

Anti_CD20_scFv_IgGFc Anchored Human T Lymphocytes Induced Raji Cells Apoptosis

PAN Zhi_yong, TAN Ying_xia,YU Kang, WU Jian_bo,ZHANG Sheng_hui,HAN Yi_xiang   

  1. Central Laboratory of Biology, Wenzhou Medical College, Wenzhou
  • Received:2006-10-08 Revised:2007-03-03 Online:2007-07-30 Published:2007-07-30

摘要: 背景与目的: 研究重组anti_CD20_scFv_IgGFc 的pLNCX质粒转染T淋巴细胞后,用锚定的T细胞杀伤 Raji细胞,观察其诱导Raji细胞凋亡的能力。 材料与方法: 重组质粒通过脂质体转染至逆转录病毒包装的PA317细胞中,600 μg/ml的G418筛选3周后,用PA317细胞培养液感染的外周血T细胞和T细胞分别去杀伤等量的Raji细胞,于不同时间点用流式细胞仪检测Raji细胞AnnexinⅤ和Bcl_2的阳性率。 结果: 流式细胞仪检测发现在24 h内,CD20阳性的Raji细胞上AnnexinⅤ阳性率明显高于对照组,Bcl_2的阳性表达率较对照组明显降低。 结论: 含 anti_CD20_scFv_IgGFc重组子基因修饰的T细胞在数小时内就可引起Raji细胞发生凋亡,其早期凋亡可能通过启动AnnexinⅤ,同时通过抑制Bcl_2的表达而加快Raji细胞凋亡达到靶向杀伤的作用。

关键词: 淋巴瘤, CD20, 免疫治疗

Abstract: BACKGROUND & AIM: By transfecting the CD20_scFv_IgGFc recombinant pLNCX plasmid into T lymphocytes and employing the anchored T lymphocytes to attack the Raji lymphoma cells in vitro, to explore the capability of the anchored T lymphocytes in inducing apoptosis of Raji cells. MATERIALS AND METHODS: Recombinant plasmid was transfected into retrovirus_packed PA317 cell lines with lipofectamine reagent, and then the growth of the trancfectants were selected in a medium supplemented with G418(600 μg/ml) for 3 weeks. Raji cells were attacked by T lymphocytes with and without transfected PA317 supernatant. The AnnexinⅤ and Bcl_2 positive rates in Raji cells at different times were monitored by flow cytometry. RESULTS: The expressions of Annexin V and Bcl_2 in CD20+ Raji cells were obviously different to the control group within 24 hours. CONCLUSION: Anti_CD20_scFv_IgGFc modified T cells could provoke Raji cells apoptosis within several hours. AnnexinⅤ may initiate Raji early apoptosis, and meanwhile genetically modified T cells could inhibit Bcl_2 expression of Raji cells to its speed up its apoptosis.

Key words: lymphoma, CD20, immunotherapy