细胞内递送的抗蓖麻毒素A链抗体对蓖麻毒素诱导的细胞毒性的保护作用。
Protective effects of anti-ricin A-chain antibodies delivered intracellularly against ricin-induced cytotoxicity.
作者信息
Wu Feng, Fan Shaoan, Martiniuk Frank, Pincus Seth, Müller Sybille, Kohler Heinz, Tchou-Wong Kam-Meng
机构信息
Feng Wu, Shaoan Fan, Kam-Meng Tchou-Wong, Department of Environmental Medicine, New York University School of Medicine, 57 Old Forge Road, Tuxedo, NY 10987, United States.
出版信息
World J Biol Chem. 2010 May 26;1(5):188-95. doi: 10.4331/wjbc.v1.i5.188.
AIM
To evaluate the ability of anti-ricin A-chain antibodies, delivered intracellularly, to protect against ricin-induced cytotoxicity in RAW264.7 cells.
METHODS
Anti-deglycosylated ricin A-chain antibody and RAC18 anti-ricin A-chain monoclonal antibody were delivered intracellularly by encapsulating in liposomes or via conjugation with the cell-penetrating MTS-transport peptide. RAW264.7 cells were incubated with these antibodies either before or after ricin exposure. The changes in cytotoxicity were estimated by MTT assay. Co-localization of internalized antibody and ricin was evaluated by fluorescence microscopy.
RESULTS
Internalized antibodies significantly increased cell viability either before or after ricin exposure compared to the unconjugated antibodies. Fluorescence microscopy confirmed the co-localization of internalized antibodies and ricin inside the cells.
CONCLUSION
Intracellular delivery of antibodies to neutralize the ricin toxin after cellular uptake supports the potential use of cell-permeable antibodies for post-exposure treatment of ricin intoxication.
目的
评估细胞内递送的抗蓖麻毒素A链抗体对RAW264.7细胞中蓖麻毒素诱导的细胞毒性的防护能力。
方法
通过脂质体包封或与细胞穿透性MTS转运肽偶联，将抗去糖基化蓖麻毒素A链抗体和RAC18抗蓖麻毒素A链单克隆抗体递送至细胞内。在蓖麻毒素暴露之前或之后，用这些抗体孵育RAW264.7细胞。通过MTT法评估细胞毒性的变化。通过荧光显微镜评估内化抗体与蓖麻毒素的共定位。
结果
与未偶联的抗体相比，内化抗体在蓖麻毒素暴露之前或之后均显著提高了细胞活力。荧光显微镜证实了细胞内内化抗体与蓖麻毒素的共定位。
结论
细胞摄取后细胞内递送抗体以中和蓖麻毒素毒素，支持了可渗透细胞的抗体在蓖麻毒素中毒暴露后治疗中的潜在应用。
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