Department of Chemistry, Biotechnology and Chemical Engineering, Graduate School of Science and Engineering , Kagoshima University , 1-21-40 Korimoto , Kagoshima 890-0065 , Japan.
Department of Molecular Enzymology, Faculty of Life Sciences , Kumamoto University , 1-1-1 Honjo, Chuo-ku , Kumamoto 860-8556 , Japan.
Bioconjug Chem. 2019 Nov 20;30(11):2811-2821. doi: 10.1021/acs.bioconjchem.9b00560. Epub 2019 Oct 9.
Adjuvants enhance the immune response during vaccination. Among FDA-approved adjuvants, aluminum salts are most commonly used in vaccines. Although aluminum salts enhance humoral immunity, they show a limited effect for cell-mediated immune responses. Thus, further development of adjuvants that induce T-cell-mediated immune response is needed. Toll-like receptors (TLRs) recognizing specific pathogen-associated molecular patterns activate innate immunity, which is crucial to shape adaptive immunity. Using TLR ligands as novel adjuvants in vaccines has therefore attracted substantial attention. Among them a small molecule TLR7 ligand, imiquimod, has been approved for clinical use, but its use is restricted to local administration due to unwanted adverse side effects when used systematically. Since TLR7 is mainly located in the endosomal compartment of immune cells, efficient transport of the ligand into the cells is important for improving the potency of the TLR7 ligand. In this study we examined gold nanoparticles (GNPs) immobilized with α-mannose as carriers for a TLR7 ligand to target immune cells. The small molecule synthetic TLR7 ligand, 2-methoxyethoxy-8-oxo-9-(4-carboxy benzyl)adenine (1V209), and α-mannose were coimmobilized via linker molecules consisting of thioctic acid on the GNP surface (1V209-αMan-GNPs). The in vitro cytokine production activity of 1V209-αMan-GNPs was higher than that of the unconjugated 1V209 derivative in mouse bone marrow-derived dendritic cells and in human peripheral blood mononuclear cells. In the in vivo immunization study, 1V209-αMan-GNPs induced significantly higher titers of IgG2c antibody specific to ovalbumin as an antigen than did unconjugated 1V209, and splenomegaly and weight loss were not observed. These results indicate that 1V209-αMan-GNPs could be useful as safe and effective adjuvants for development of vaccines against infectious diseases and cancer.
佐剂增强疫苗接种时的免疫反应。在 FDA 批准的佐剂中,铝盐最常用于疫苗。虽然铝盐增强体液免疫,但它们对细胞介导的免疫反应显示出有限的作用。因此,需要进一步开发诱导 T 细胞介导的免疫反应的佐剂。识别特定病原体相关分子模式的 Toll 样受体 (TLR) 激活先天免疫,这对于塑造适应性免疫至关重要。因此,使用 TLR 配体作为疫苗中的新型佐剂引起了广泛关注。其中,小分子 TLR7 配体咪喹莫特已被批准用于临床,但由于系统性使用时会产生不良的不良反应,其使用仅限于局部给药。由于 TLR7 主要位于免疫细胞的内体隔室中,因此将配体有效递送到细胞中对于提高 TLR7 配体的效力很重要。在这项研究中,我们研究了用 α-甘露糖固定化的金纳米粒子 (GNPs) 作为 TLR7 配体的载体来靶向免疫细胞。小分子合成 TLR7 配体 2-甲氧基乙氧基-8-氧代-9-(4-羧基苄基)腺嘌呤 (1V209) 和 α-甘露糖通过硫辛酸连接分子共固定在 GNP 表面 (1V209-αMan-GNPs)。与未缀合的 1V209 衍生物相比,1V209-αMan-GNPs 在小鼠骨髓来源的树突状细胞和人外周血单核细胞中的细胞因子产生活性更高。在体内免疫研究中,1V209-αMan-GNPs 诱导的针对卵清蛋白作为抗原的 IgG2c 抗体的效价明显高于未缀合的 1V209,并且未观察到脾肿大和体重减轻。这些结果表明,1V209-αMan-GNPs 可用作安全有效的佐剂,用于开发针对传染病和癌症的疫苗。
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