Jiangsu Key Laboratory for High-Tech Research and Development of Veterinary Biopharmaceuticals, Jiangsu Agri-Animal Husbandry Vocational College, Taizhou, 225300, Jiangsu Province, People's Republic of China.
Discipline: Microbiology, School of Life Sciences, College of Agriculture, Engineering and Science, University of KwaZulu-Natal, Durban, 4000, South Africa.
Sci Rep. 2022 Jul 15;12(1):12122. doi: 10.1038/s41598-022-16378-x.
Dendritic cells (DCs) are the antigen-presenting cells that initiate and direct adaptive immune responses, and thus are critically important in vaccine design. Although DC-targeting vaccines have attracted attention, relevant studies on chicken are rare. A high diversity T7 phage display nanobody library was constructed for bio-panning of intact chicken bone marrow DCs to find DC-specific binding nanobodies. After three rounds of screening, 46 unique sequence phage clones were identified from 125 randomly selected phage clones. Several DC-binding phage clones were selected using the specificity assay. Phage-54, -74, -16 and -121 bound not only with chicken DCs, but also with duck and goose DCs. In vitro, confocal microscopy observation demonstrated that phage-54 and phage-74 efficiently adsorbed onto DCs within 15 min compared to T7-wt. The pull-down assay, however, did not detect any of the previously reported proteins for chicken DCs that could have interacted with the nanobodies displayed on phage-54 and phage-74. Nonetheless, Specified pathogen-free chickens immunized with phage-54 and phage-74 displayed higher levels of anti-p10 antibody than the T7-wt, indicating enhanced antibody production by nanobody mediated-DC targeting. Therefore, this study identified two avian (chicken, duck and goose) DC-specific binding nanobodies, which may be used for the development of DC-targeting vaccines.
树突状细胞 (DCs) 是启动和指导适应性免疫反应的抗原呈递细胞，因此在疫苗设计中至关重要。虽然针对 DC 的疫苗引起了关注，但关于鸡的相关研究很少。为了从完整的鸡骨髓 DC 中进行生物淘选以寻找 DC 特异性结合纳米抗体，构建了一个高多样性的 T7 噬菌体展示纳米抗体文库。经过三轮筛选，从 125 个随机选择的噬菌体克隆中鉴定出 46 个独特序列的噬菌体克隆。使用特异性测定法从噬菌体中选择了几个 DC 结合的噬菌体克隆。噬菌体 54、74、16 和 121 不仅与鸡 DC 结合，而且与鸭和鹅的 DC 结合。在体外，共聚焦显微镜观察表明，与 T7-wt 相比，噬菌体 54 和噬菌体 74 在 15 分钟内更有效地吸附到 DC 上。然而，下拉测定法未检测到与噬菌体 54 和噬菌体 74 上展示的纳米抗体相互作用的任何先前报道的鸡 DC 蛋白。尽管如此，用噬菌体 54 和噬菌体 74 免疫 SPF 鸡显示出比 T7-wt 更高水平的抗 p10 抗体，表明纳米抗体介导的 DC 靶向增强了抗体产生。因此，本研究鉴定了两种禽类（鸡、鸭和鹅）DC 特异性结合纳米抗体，可用于开发针对 DC 的疫苗。
