Targeting of the C-Type Lectin Receptor Langerin Using Bifunctional Mannosylated Antigens
Targeting of the C-Type Lectin Receptor Langerin Using Bifunctional Mannosylated Antigens
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DOI：
10.3389/fcell.2020.00556
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发表时间：
2020-07-14
影响因子：
5.5
通讯作者：
van Kooyk, Yvette
中科院分区：
文献类型：
--
作者：
Li, Rui-Jun Eveline;Hogervorst, Tim P.;van Kooyk, Yvette
朗格汉斯细胞 (LC) 是存在于皮肤中的抗原呈递细胞。它们独特地表达高水平的 C 型凝集素受体 Langerin (CD207)，这是癌症免疫治疗疫苗接种策略中抗原递送的一个有吸引力的靶点。我们在这里评估了一个由 20 个合成的、明确的甘露糖苷簇组成的文库，这些簇由六种单甘露糖苷、二甘露糖苷或三甘露糖苷中的一种、两种和三种组成，附加到寡肽主链上，使用表面等离子体共振和流式细胞术定量与 Langerin 结合。发现Langerin结合亲和力随着甘露糖苷数量的增加而增加。甘露糖苷的六价呈递导致3至12μM范围内的结合亲和力。二甘露糖苷和三甘露糖苷的三价呈递导致相同范围内的Langerin亲和力。随后模型黑色素瘤 gp100 抗原肽配备有二甘露糖苷和三甘露糖苷的六价簇作为靶向部分。令人惊讶的是，尽管双功能缀合物以 Langerin 依赖性方式被 LC 吸收，但观察到细胞毒性 T 细胞的抗原呈递有限。这些结果表明，免疫治疗疫苗上的靶向聚糖部分不仅应该验证靶标结合，还应该验证对生物学的持续影响，例如抗原呈递给 CD8(+) 和 CD4(+)T 细胞。
Langerhans cells (LCs) are antigen-presenting cells that reside in the skin. They uniquely express high levels of the C-type lectin receptor Langerin (CD207), which is an attractive target for antigen delivery in immunotherapeutic vaccination strategies against cancer. We here assess a library of 20 synthetic, well-defined mannoside clusters, built up from one, two, and three of six monomannosides, dimannosides, or trimannosides, appended to an oligopeptide backbone, for binding with Langerin using surface plasmon resonance and flow cytometric quantification. It is found that Langerin binding affinity increases with increasing number of mannosides. Hexavalent presentation of the mannosides resulted in binding affinities ranging from 3 to 12 mu M. Trivalent presentation of the dimannosides and trimannosides led to Langerin affinity in the same range. The model melanoma gp100 antigenic peptide was subsequently equipped with a hexavalent cluster of the dimannosides and trimannosides as targeting moieties. Surprisingly, although the bifunctional conjugates were taken up in LCs in a Langerin-dependent manner, limited antigen presentation to cytotoxic T cells was observed. These results indicate that targeting glycan moieties on immunotherapeutic vaccines should not only be validated for target binding, but also on the continued effects on biology, such as antigen presentation to both CD8(+)and CD4(+)T cells.