Differential cytosolic delivery and presentation of antigen by listeriolysin O-liposomes to macrophages and dendritic cells.
Differential cytosolic delivery and presentation of antigen by listeriolysin O-liposomes to macrophages and dendritic cells.
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DOI：
10.1021/mp049896v
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发表时间：
2005
影响因子：
4.9
通讯作者：
Ethan M. Stier;M. Mandal;Kyung-Dall Lee
中科院分区：
文献类型：
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作者：
Ethan M. Stier;M. Mandal;Kyung-Dall Lee
将抗原蛋白递送至抗原呈递细胞（APC）（如巨噬细胞（MPhi）和树突状细胞（DC））的胞质溶胶是有效的CD 8 T细胞介导的免疫应答所需的。我们以前已经表明，共同封装的抗原蛋白内的pH敏感的脂质体与嗜酸乳杆菌溶血素O（LLO），单核细胞增生李斯特菌的成孔蛋白，产生有效的主要组织相容性复合物I类（MHC I）限制性免疫反应，在体外和体内。在这项研究中，我们试图分析两个重要的APC，巨噬细胞和树突状细胞中的LLO介导的脂质体抗原的胞质递送的相对效率，通过检查在培养的小鼠骨髓来源的MPhis（BMMPhis）和DC（BMDCs）中抗原呈递给T细胞的顺序步骤。在与细胞孵育的给定浓度的脂质体抗原下，BMMPhis总体上比BMDCs更好地呈递脂质体抗原，并且在通过抗原摄取使呈递标准化后也观察到这种趋势。然而，当可溶性抗原被直接引入到胞质溶胶中时，BMDCs比BMMPhis更有效地呈递抗原。此外，当APC外部负载蛋白质的抗原肽时，BMDC显示出更高水平的细胞表面MHC I-肽复合物，并且比BMMPhis更有效地呈递肽。这些结果结合在一起表明，LLO介导的脂质体抗原从内体/溶酶体区室的释放在BMMPhis中可能比在BMDC中更明显，并进一步暗示LLO的差异活性和不同抗原呈递细胞类型中LLO介导的内体逃逸的不同效率。
Delivery of antigenic protein to the cytosol of antigen-presenting cells (APCs), such as macrophages (MPhi) and dendritic cells (DCs), is required for an efficient CD8 T-cell-mediated immune response. We have previously shown that co-encapsulation of antigenic protein inside pH-sensitive liposomes with listeriolysin O (LLO), a pore-forming protein of Listeria monocytogenes, generates efficient major histocompatibility complex class I (MHC I)-restricted immune responses both in vitro and in vivo. In this study, we sought to analyze the relative efficiency of LLO-mediated cytosolic delivery of liposomal antigen in two important APCs, macrophages and dendritic cells, by examining the sequential steps involved in antigen presentation to T-cells in cultured mouse bone marrow-derived MPhis (BMMPhis) and DCs (BMDCs). BMMPhis overall presented liposomal antigen better than BMDCs at a given concentration of liposomal antigen incubated with cells, and the trend was also observed after the presentation was normalized by the uptake of antigen. When soluble antigen was directly introduced into the cytosol, however, BMDCs presented the antigen more efficiently than BMMPhis. In addition, when the APCs were externally loaded with the antigenic peptide of the protein, BMDCs displayed a higher level of cell surface MHC I-peptide complexes and presented the peptide more efficiently than BMMPhis. These results combined together suggest that LLO-mediated release of liposomal antigen from the endosomal/lysosomal compartment may be more pronounced in BMMPhis than in BMDCs, and further implicates differential activity of LLO and varying efficiency of LLO-mediated endosomal escape in different antigen-presenting cell types.