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用负载乙肝表面抗原的弹性脂质体脉冲处理的小鼠树突状细胞对摄取及免疫应答产生的评估

Evaluation of uptake and generation of immune response by murine dendritic cells pulsed with hepatitis B surface antigen-loaded elastic liposomes.

作者信息

Mishra Dinesh, Mishra Pradyumna Kumar, Dubey Vaibhav, Dabadghao Sunil, Jain Narendra Kumar

机构信息

Department of Pharmaceutical Sciences, Dr. H.S. Gour University, Sagar, India.

出版信息

Vaccine. 2007 Sep 28;25(39-40):6939-44. doi: 10.1016/j.vaccine.2007.06.055. Epub 2007 Jul 17.

Abstract

Hepatitis B surface antigen (HBsAg)-loaded elastic liposomes were studied for qualitative and quantitative uptake in vitro by murine dendritic cells (DCs) generated from bone marrow mononuclear cells. Internalization of the vesicles by the DCs was documented using fluorescence microscopy. Kinetics of uptake of antigen-loaded elastic vesicles by the DCs studied through flow cytometry showed a peak uptake at 6h. The ability of the antigen pulsed DCs to stimulate autologous peripheral blood lymphocytes was demonstrated by BrdU assay. Further evaluation by multiplex cytometric bead array analysis demonstrated a predominantly TH1 type of immune response. Our results suggest that HBsAg-loaded elastic vesicles as antigen delivery module and DCs as antigen presenting cells are able to generate a protective immune response. The property of elastic liposomes to traverse and target the immunological milieu of the skin makes it an attractive vehicle for development of a transcutaneous vaccine against hepatitis B virus.

摘要

对负载乙肝表面抗原(HBsAg)的弹性脂质体进行了研究,以观察其在体外被由骨髓单核细胞生成的小鼠树突状细胞(DCs)进行定性和定量摄取的情况。利用荧光显微镜记录了DCs对这些囊泡的内化过程。通过流式细胞术研究DCs摄取负载抗原的弹性囊泡的动力学,结果显示在6小时时摄取量达到峰值。通过BrdU检测证明了经抗原脉冲处理的DCs刺激自体外周血淋巴细胞的能力。通过多重细胞计数微球阵列分析进一步评估表明,主要产生了TH1型免疫反应。我们的结果表明,负载HBsAg的弹性囊泡作为抗原递送模块,DCs作为抗原呈递细胞,能够产生保护性免疫反应。弹性脂质体穿越并靶向皮肤免疫环境的特性使其成为开发抗乙肝病毒经皮疫苗的有吸引力的载体。

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