鼻咽癌(Nasopharyngeal carcinoma,NPC)与EB病毒(Epstein-Barr virus,EBV)感染密切相关。鼻咽癌细胞中表达的EB病毒潜伏膜蛋白2(Latent membrane protein 2,LMP2)是免疫疗法的理想靶抗原。靶向EBV-LMP2疫苗已被证明是安全有效的,可在NP...鼻咽癌(Nasopharyngeal carcinoma,NPC)与EB病毒(Epstein-Barr virus,EBV)感染密切相关。鼻咽癌细胞中表达的EB病毒潜伏膜蛋白2(Latent membrane protein 2,LMP2)是免疫疗法的理想靶抗原。靶向EBV-LMP2疫苗已被证明是安全有效的,可在NPC患者体内增强特异性细胞毒性T淋巴细胞(Cytotoxic T lymphocyte,CTL)应答。目前面临的挑战是如何提高NPC患者的临床反应率,其关键在于阐明LMP2特异性CTL的细胞毒性。为研究rAd-LMP2疫苗诱导的LMP2A特异性CTL对肿瘤细胞的杀伤效果,本研究利用rMVA-LMP2-GFP感染P815细胞(H-2d)构建表达EBV-LMP2A和GFP蛋白的P815-LMP2-GFP靶细胞,用rAd-LMP2疫苗免疫BALB/c小鼠(H-2d)来诱生LMP2A特异性的CTL细胞作为效应细胞。然后,应用活细胞成像技术观察LMP2A特异性CTL对P815-LMP2-GFP靶细胞的杀伤过程。结果显示成功构建了P815-LMP2-GFP靶细胞和LMP2A特异性CTL效应细胞,并以成像的方式实时动态记录了LMP2A特异性CTL识别并杀死靶细胞的过程。P815-LMP2-GFP靶细胞可在2h内被LMP2A特异性CTL识别并杀伤。本研究建立了一种直接监测LMP2A特异性CTL识别并杀伤靶细胞过程的动态可视化方法。因此,本研究为肿瘤疫苗的体外抗肿瘤免疫效果的研究提供了新手段和新思路。展开更多
Objective To evaluate the safety and effectiveness of a vaccine based on latent membrane protein 2(LMP2)modified dendritic cells(DCs)that boosts specific responses of cytotoxic T lymphocytes(CTLs)to LMP2 before and af...Objective To evaluate the safety and effectiveness of a vaccine based on latent membrane protein 2(LMP2)modified dendritic cells(DCs)that boosts specific responses of cytotoxic T lymphocytes(CTLs)to LMP2 before and after intradermal injection in patients with nasopharyngeal carcinoma(NPC).Methods DCs were derived from peripheral blood monocytes of patients with NPC.We prepared LMP2-DCs infected by recombinant adenovirus vector expressing LMP2(rAd-LMP2).NPC patients were immunized with 2×105 LMP2-DCs by intradermal injection at week 0 and after the second and fourth weeks.Specific responses to LMP2 were detected by enzyme-linked immunospot(ELISPOT)assay at week 0 and at the fifth and eighth weeks.Local clinicians performed the follow-up and tracking of patients.Results We demonstrated that DCs derived from monocytes displayed typical DC morphologies;the expression of LMP2 in the LMP2-DCs vaccine was confirmed by immunocytochemical assay.Twenty-nine patients with NPC were enrolled in this clinical trial.The LMP2-DCs vaccine was well tolerated in all of the patients.Boosted responses to LMP2 peptide sub-pools were observed in 18 of the 29 patients with NPC.The follow-up data of 29 immunized patients from April,2010 to April 2015 indicated a five-year survival rate of 94.4%in responders and 45.5%in non-responders.Conclusion In this pilot study,we demonstrated that the LMP2-DCs vaccine is safe and effective in patients with NPC.Specific CTLs responses to LMP2 play a certain role in controlling and preventing the recurrence and metastasis of NPC,which warrants further clinical testing.展开更多
Epstein-Barr virus (EBV) infection is closely associated with nasopharyngeal carcinoma (NPC)and considered one of the major risk factors[1]. A limited number of viral proteins, such as latent membrane proteins (LMP1, ...Epstein-Barr virus (EBV) infection is closely associated with nasopharyngeal carcinoma (NPC)and considered one of the major risk factors[1]. A limited number of viral proteins, such as latent membrane proteins (LMP1, LMP2) and EB nuclear antigen1 (EBNA1), are expressed in NPC cells[2].Recognition epitopes for CD8+and CD4+T cells were included in the LMP2 antigen and EBNA1 C-terminal region (amino acid No. 380-641), respectively[3,4].Both CD4+and CD8+memory T-cell responses were efficiently reactivated by EBNA1 and LMP2[5;6].展开更多
文摘鼻咽癌(Nasopharyngeal carcinoma,NPC)与EB病毒(Epstein-Barr virus,EBV)感染密切相关。鼻咽癌细胞中表达的EB病毒潜伏膜蛋白2(Latent membrane protein 2,LMP2)是免疫疗法的理想靶抗原。靶向EBV-LMP2疫苗已被证明是安全有效的,可在NPC患者体内增强特异性细胞毒性T淋巴细胞(Cytotoxic T lymphocyte,CTL)应答。目前面临的挑战是如何提高NPC患者的临床反应率,其关键在于阐明LMP2特异性CTL的细胞毒性。为研究rAd-LMP2疫苗诱导的LMP2A特异性CTL对肿瘤细胞的杀伤效果,本研究利用rMVA-LMP2-GFP感染P815细胞(H-2d)构建表达EBV-LMP2A和GFP蛋白的P815-LMP2-GFP靶细胞,用rAd-LMP2疫苗免疫BALB/c小鼠(H-2d)来诱生LMP2A特异性的CTL细胞作为效应细胞。然后,应用活细胞成像技术观察LMP2A特异性CTL对P815-LMP2-GFP靶细胞的杀伤过程。结果显示成功构建了P815-LMP2-GFP靶细胞和LMP2A特异性CTL效应细胞,并以成像的方式实时动态记录了LMP2A特异性CTL识别并杀死靶细胞的过程。P815-LMP2-GFP靶细胞可在2h内被LMP2A特异性CTL识别并杀伤。本研究建立了一种直接监测LMP2A特异性CTL识别并杀伤靶细胞过程的动态可视化方法。因此,本研究为肿瘤疫苗的体外抗肿瘤免疫效果的研究提供了新手段和新思路。
基金Mega Project of Research on the Prevention and Control of HIV/AIDS,Viral Hepatitis Infectious Diseases[Grant No:2018ZX10102001]the Key Science and Technology Program of Guangxi Zhuang Autonomous Region[Grant No.14124003-3]+1 种基金the National High Technology Research and Development Program of China[Grant No.2007AA021107]and the National Basic Research Program of China[973 Program,Grant No.2011CB504800]。
文摘Objective To evaluate the safety and effectiveness of a vaccine based on latent membrane protein 2(LMP2)modified dendritic cells(DCs)that boosts specific responses of cytotoxic T lymphocytes(CTLs)to LMP2 before and after intradermal injection in patients with nasopharyngeal carcinoma(NPC).Methods DCs were derived from peripheral blood monocytes of patients with NPC.We prepared LMP2-DCs infected by recombinant adenovirus vector expressing LMP2(rAd-LMP2).NPC patients were immunized with 2×105 LMP2-DCs by intradermal injection at week 0 and after the second and fourth weeks.Specific responses to LMP2 were detected by enzyme-linked immunospot(ELISPOT)assay at week 0 and at the fifth and eighth weeks.Local clinicians performed the follow-up and tracking of patients.Results We demonstrated that DCs derived from monocytes displayed typical DC morphologies;the expression of LMP2 in the LMP2-DCs vaccine was confirmed by immunocytochemical assay.Twenty-nine patients with NPC were enrolled in this clinical trial.The LMP2-DCs vaccine was well tolerated in all of the patients.Boosted responses to LMP2 peptide sub-pools were observed in 18 of the 29 patients with NPC.The follow-up data of 29 immunized patients from April,2010 to April 2015 indicated a five-year survival rate of 94.4%in responders and 45.5%in non-responders.Conclusion In this pilot study,we demonstrated that the LMP2-DCs vaccine is safe and effective in patients with NPC.Specific CTLs responses to LMP2 play a certain role in controlling and preventing the recurrence and metastasis of NPC,which warrants further clinical testing.
基金supported by the Mega Project of Research on the Prevention and Control of HIV/AIDS,Viral Hepatitis Infectious Diseases [2018ZX10711001]
文摘Epstein-Barr virus (EBV) infection is closely associated with nasopharyngeal carcinoma (NPC)and considered one of the major risk factors[1]. A limited number of viral proteins, such as latent membrane proteins (LMP1, LMP2) and EB nuclear antigen1 (EBNA1), are expressed in NPC cells[2].Recognition epitopes for CD8+and CD4+T cells were included in the LMP2 antigen and EBNA1 C-terminal region (amino acid No. 380-641), respectively[3,4].Both CD4+and CD8+memory T-cell responses were efficiently reactivated by EBNA1 and LMP2[5;6].
