目的:探讨红景天苷(salidroside,SAL)对树突状细胞(dendritic cell,DC)表型及细胞毒性T细胞(cytotoxic T lymphocyte,CTL)抗肿瘤能力的影响。方法:选用Lewis肺癌细胞株3LL、野生型C57BL/6和TLR4-/-C57BL/6小鼠,获取小鼠骨髓来源的DC前...目的:探讨红景天苷(salidroside,SAL)对树突状细胞(dendritic cell,DC)表型及细胞毒性T细胞(cytotoxic T lymphocyte,CTL)抗肿瘤能力的影响。方法:选用Lewis肺癌细胞株3LL、野生型C57BL/6和TLR4-/-C57BL/6小鼠,获取小鼠骨髓来源的DC前体细胞,经过培养分化成未成熟DC,收获第6天的DC,经磁珠分选后获得纯度较高的CD11c+DC。将细胞分成PBS组、SAL组和脂多糖(lipopolysaccharide,LPS)组,培养48 h后用流式细胞术检测SAL体外对DC表面分子、吞噬功能、TLR4通路和对T细胞杀伤能力的影响。结果:与PBS组比较,SAL组DC的表面分子CD80、CD86、MHCⅡ表达水平显著升高(均P<0.05)、吞噬功能显著下降(P<0.05)、TLR4表达水平显著升高(P<0.01);与野生型组比较,TLR4-/-组DC经SAL或LPS处理后,其表面分子CD80、CD86、MHCⅡ的表达水平显著降低(均P<0.05);与PBS组比较,SAL组刺激活化的CTL对肺癌3LL细胞的杀伤效应显著升高(P<0.05)。结论:SAL可以通过调控TLR4诱导DC成熟,从而提高T细胞的杀伤能力。展开更多
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
Equine herpesvirus-1 (EHV-1) remains one of the most common viral pathogens affecting horses worldwide presenting as a persistent infection which can establish latency in nerve ganglia (trigeminal ganglion), lymphoid ...Equine herpesvirus-1 (EHV-1) remains one of the most common viral pathogens affecting horses worldwide presenting as a persistent infection which can establish latency in nerve ganglia (trigeminal ganglion), lymphoid tissues of the respiratory tract and peripheral blood lymphocytes. EHV-1 infection induces both humoral and cellular immune responses in horses. Virus neutralising antibody, particularly in the nasopharynx, is to kill free virus shed from infected epithelial cells. Hence this antibody has important functions in reducing virus shedding and spreading infection to cohorts. Cellular immune responses, particularly those carried out by cytotoxic T lymphocyte (CTL), have been shown to be effective in killing virus-infected cells in vitro. This review underlines the state of knowledge regarding immunity to EHV-1 and also its interaction with equine lymphocyte. Finally, the review also includes the importance of the viral immediate early (IE) protein in the pathogenesis of EHV-1. This information can be used as the basis for future research.展开更多
Objective: To established methods for cryopreserving peripheral blood mononuclear cells (PBMCs) andproducing DCs from cryopreserved PBMCs. Methods:Mature DCs were generated from cryopreserved PBMCs by using IL-4, GM...Objective: To established methods for cryopreserving peripheral blood mononuclear cells (PBMCs) andproducing DCs from cryopreserved PBMCs. Methods:Mature DCs were generated from cryopreserved PBMCs by using IL-4, GM-CSF, TNF-a, IL-1b, IL-6, pgE2 and LPS. The phenotype of the resultant DCs was investigated by flow cytometry. The functions of the resultant DCs were verified by Elispot assay. Results: The resultant DCs expressed high levels of HLA ABC, HLA DR, costimulatory molecules and the DC maturation marker CD83. The mature DCs wegenerated from frozen PBMCs were able to prime CD8 T cells into long term IFN-g producing peptide specific CTL. Conclusion: The DCs we developed from cryopreservedPBMC were fully mature and had the capability tostimulate immune reaction. Thus, we developed a method to generate functional mature DC from cryopreservedPBMC.展开更多
文摘目的:探讨红景天苷(salidroside,SAL)对树突状细胞(dendritic cell,DC)表型及细胞毒性T细胞(cytotoxic T lymphocyte,CTL)抗肿瘤能力的影响。方法:选用Lewis肺癌细胞株3LL、野生型C57BL/6和TLR4-/-C57BL/6小鼠,获取小鼠骨髓来源的DC前体细胞,经过培养分化成未成熟DC,收获第6天的DC,经磁珠分选后获得纯度较高的CD11c+DC。将细胞分成PBS组、SAL组和脂多糖(lipopolysaccharide,LPS)组,培养48 h后用流式细胞术检测SAL体外对DC表面分子、吞噬功能、TLR4通路和对T细胞杀伤能力的影响。结果:与PBS组比较,SAL组DC的表面分子CD80、CD86、MHCⅡ表达水平显著升高(均P<0.05)、吞噬功能显著下降(P<0.05)、TLR4表达水平显著升高(P<0.01);与野生型组比较,TLR4-/-组DC经SAL或LPS处理后,其表面分子CD80、CD86、MHCⅡ的表达水平显著降低(均P<0.05);与PBS组比较,SAL组刺激活化的CTL对肺癌3LL细胞的杀伤效应显著升高(P<0.05)。结论:SAL可以通过调控TLR4诱导DC成熟,从而提高T细胞的杀伤能力。
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。
文摘Equine herpesvirus-1 (EHV-1) remains one of the most common viral pathogens affecting horses worldwide presenting as a persistent infection which can establish latency in nerve ganglia (trigeminal ganglion), lymphoid tissues of the respiratory tract and peripheral blood lymphocytes. EHV-1 infection induces both humoral and cellular immune responses in horses. Virus neutralising antibody, particularly in the nasopharynx, is to kill free virus shed from infected epithelial cells. Hence this antibody has important functions in reducing virus shedding and spreading infection to cohorts. Cellular immune responses, particularly those carried out by cytotoxic T lymphocyte (CTL), have been shown to be effective in killing virus-infected cells in vitro. This review underlines the state of knowledge regarding immunity to EHV-1 and also its interaction with equine lymphocyte. Finally, the review also includes the importance of the viral immediate early (IE) protein in the pathogenesis of EHV-1. This information can be used as the basis for future research.
文摘Objective: To established methods for cryopreserving peripheral blood mononuclear cells (PBMCs) andproducing DCs from cryopreserved PBMCs. Methods:Mature DCs were generated from cryopreserved PBMCs by using IL-4, GM-CSF, TNF-a, IL-1b, IL-6, pgE2 and LPS. The phenotype of the resultant DCs was investigated by flow cytometry. The functions of the resultant DCs were verified by Elispot assay. Results: The resultant DCs expressed high levels of HLA ABC, HLA DR, costimulatory molecules and the DC maturation marker CD83. The mature DCs wegenerated from frozen PBMCs were able to prime CD8 T cells into long term IFN-g producing peptide specific CTL. Conclusion: The DCs we developed from cryopreservedPBMC were fully mature and had the capability tostimulate immune reaction. Thus, we developed a method to generate functional mature DC from cryopreservedPBMC.
