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,600μg/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Annexin Ⅴ和Bcl-2的阳性率。结果:流式细胞仪检测发现在24h内,CD20阳性的Raji细胞上Annexin Ⅴ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Annexin Ⅴ,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。
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（600μg/ml） for 3 weeks. Raji cells were attacked by T lymphocytes with and without transfected PA317 supematant. The Annexin V 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. Annexin V 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.
出处
《癌变．畸变．突变》
CAS
CSCD
2007年第4期290-293,共4页
Carcinogenesis,Teratogenesis & Mutagenesis
基金
浙江省自然科学基金项目(302781)
