目的:EB病毒与鼻咽癌的发生有密切的关系,而其编码的潜伏膜蛋白1(latent membrane protein 1,LMP1)被证实具有转化致瘤的作用。本实验的目的是观察LMP1是否使细胞产生对顺铂的抗性,并探索其中的发生机制。方法:用脂质体转染带有LMP1的...目的:EB病毒与鼻咽癌的发生有密切的关系,而其编码的潜伏膜蛋白1(latent membrane protein 1,LMP1)被证实具有转化致瘤的作用。本实验的目的是观察LMP1是否使细胞产生对顺铂的抗性,并探索其中的发生机制。方法:用脂质体转染带有LMP1的质粒进入CNE2细胞,G418筛选构成稳定株,qRT-PCR和western检测其LMP1的表达情况。MTT和Annexin V/PI双染实验检测细胞对顺铂的敏感性。结果:CNE2/LMP1细胞能够稳定表达LMP1。LMP1能够促进细胞抵抗顺铂的生长抑制作用和抗顺铂诱导的凋亡。结论:EB病毒潜伏膜蛋白LMP1增强细胞对顺铂的抗性。展开更多
T cells modified with chimeric antigen receptor are an attractive strategy to treat Epstein-Barr virus（EBV） associated malignancies.The EBV latent membrane protein 1（LMP1） is a 66-KD integral membrane protein enco...T cells modified with chimeric antigen receptor are an attractive strategy to treat Epstein-Barr virus（EBV） associated malignancies.The EBV latent membrane protein 1（LMP1） is a 66-KD integral membrane protein encoded by EBV that consists of transmembrane-spanning loops.Previously,we have identified a functional signal chain variable fragment（scFv） that specifically recognizes LMP1 through phage library screening.Here,we constructed a LMP1 specific chimeric antigen receptor containing anti-LMP1 scFv,the CD28 signalling domain,and the CD3ζchain（HELA/CAR）.We tested its functional ability to target LMP1 positive nasopharyngeal carcinoma cells.HELA/CAR cells were efficiently generated using lentivirus vector encoding the LMP1-specific chimeric antigen receptor to infect activated human CD3＋ T cells.The HELA/CAR T cells displayed LMP1 specific cytolytic action and produced IFN-γ and IL-2 in response to nasopharyngeal carcinoma cells overexpressing LMP1.To demonstrate in vivo anti-tumor activity,we tested the HELA/CAR T cells in a xenograft model using an LMP1 overexpressing tumor.Intratumoral injection of anti-LMP1 HELA/CAR-T cells significantly reduced tumor growth in vivo.These results show that targeting LMP1 using HELA/CAR cells could represent an alternative therapeutic approach for patients with EBV-positive cancers.展开更多
文摘目的:EB病毒与鼻咽癌的发生有密切的关系,而其编码的潜伏膜蛋白1(latent membrane protein 1,LMP1)被证实具有转化致瘤的作用。本实验的目的是观察LMP1是否使细胞产生对顺铂的抗性,并探索其中的发生机制。方法:用脂质体转染带有LMP1的质粒进入CNE2细胞,G418筛选构成稳定株,qRT-PCR和western检测其LMP1的表达情况。MTT和Annexin V/PI双染实验检测细胞对顺铂的敏感性。结果:CNE2/LMP1细胞能够稳定表达LMP1。LMP1能够促进细胞抵抗顺铂的生长抑制作用和抗顺铂诱导的凋亡。结论:EB病毒潜伏膜蛋白LMP1增强细胞对顺铂的抗性。
基金supported in part by grants from the Special Fund of Clinical Medicine in Jiangsu Province(BL2013038)the Graduate Student Innovation Fund(CXZZ12_0563)
文摘T cells modified with chimeric antigen receptor are an attractive strategy to treat Epstein-Barr virus（EBV） associated malignancies.The EBV latent membrane protein 1（LMP1） is a 66-KD integral membrane protein encoded by EBV that consists of transmembrane-spanning loops.Previously,we have identified a functional signal chain variable fragment（scFv） that specifically recognizes LMP1 through phage library screening.Here,we constructed a LMP1 specific chimeric antigen receptor containing anti-LMP1 scFv,the CD28 signalling domain,and the CD3ζchain（HELA/CAR）.We tested its functional ability to target LMP1 positive nasopharyngeal carcinoma cells.HELA/CAR cells were efficiently generated using lentivirus vector encoding the LMP1-specific chimeric antigen receptor to infect activated human CD3＋ T cells.The HELA/CAR T cells displayed LMP1 specific cytolytic action and produced IFN-γ and IL-2 in response to nasopharyngeal carcinoma cells overexpressing LMP1.To demonstrate in vivo anti-tumor activity,we tested the HELA/CAR T cells in a xenograft model using an LMP1 overexpressing tumor.Intratumoral injection of anti-LMP1 HELA/CAR-T cells significantly reduced tumor growth in vivo.These results show that targeting LMP1 using HELA/CAR cells could represent an alternative therapeutic approach for patients with EBV-positive cancers.
