A Liposomal Platform for Delivery of a Protein Antigen to Langerin-Expressing Cells
A Liposomal Platform for Delivery of a Protein Antigen to Langerin-Expressing Cells
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DOI：
10.1021/acs.biochem.9b00402
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发表时间：
2019-05-28
期刊：
影响因子：
2.9
通讯作者：
Rademacher, Christoph
中科院分区：
文献类型：
--
作者：
Schulze, Jessica;Rentzsch, Mareike;Rademacher, Christoph
皮肤是疫苗接种的有吸引力的部位，并且具有朗格汉斯细胞的密集网络，朗格汉斯细胞是表皮中抗原递送方法的主要靶标。虽然已经显示使用基于抗体的递送方法特异性靶向朗格汉斯细胞引发必要的T细胞应答，但是以基于纳米颗粒的疫苗制剂形式靶向施用颗粒抗原一直具有挑战性。我们以前报道了一种特异性靶向配体的人Langerin，C型凝集素表达的朗格汉斯细胞。该配体存在于脂质体上，并使其对朗格汉斯细胞的摄取具有高度特异性。在这里，我们显示了一个详细的研究模型细胞系中的颗粒的摄取和细胞内路由的共聚焦和活细胞成像以及流式细胞仪检测。脂质体被内化到早期内体区室中，并在晚期内体和溶酶体中积累，随后不久释放货物。此外，我们还展示了蛋白抗原的包封及其向细胞系和原代人朗格汉斯细胞的递送。这些数据进一步支持靶向脂质体颗粒用于蛋白质疫苗应用的适用性。
The skin is an attractive site for vaccination and harbors a dense network of Langerhans cells that are the prime target for antigen delivery approaches in the epidermis. While specific targeting of Langerhans cells has been shown to elicit the necessary T-cell response using antibody-based delivery approaches, the targeted administration of particulate antigens in the form of nanoparticle-based vaccine formulations has been challenging. We previously reported on a specific targeting ligand for human Langerin, a C-type lectin expressed on Langerhans cells. This ligand is presented on liposomes and renders them highly specific for the uptake by Langerhans cells. Here we show a detailed study of the uptake and intracellular routing of the particles in model cell lines by confocal and live cell imaging as well as flow cytometric assays. Liposomes are internalized into early endosomal compartments and accumulate in late endosomes and lysosomes, shortly followed by a release of the cargo. Furthermore, we show the encapsulation of protein antigens and their delivery to cell lines and primary human Langerhans cells. These data further support the applicability of the targeted liposomal particles for protein vaccine applications.