Department of Ophthalmology, Affiliated Hospital of Qingdao University, Qingdao, People's Republic of China.
College of Food Science and Engineering, Ocean University of China, Qingdao, People's Republic of China.
Int J Nanomedicine. 2022 Feb 2;17:537-551. doi: 10.2147/IJN.S338974. eCollection 2022.
To prepare a nanobody specific to dectin 1 and verify its specificity and anti-inflammatory effects on keratitis.
The nanobody was selected from a high-quality shark-antibody library constructed with phage-display technology. The nanobody was developed in the expression systems of . Indirect ELISA was used to determine the specificity of the nanobody to recombinant dectin 1 protein. The potential of the nanobody to be recognized and expressed on the surfaces of cells and corneas was detected by immunofluorescence, and its anti-inflammatory effect on keratitis was further verified. After infection with , eyes of C57B L/6 mice were treated with nanobodies. Human corneal epithelial cells (HCECs) were pretreated with nanobodies and then incubated with . Clinical scores and slit-lamp photography were used to assess disease response in mouse corneas. RT-PCR and ELISA were used to evaluate mRNA and protein expression of IL1β and IL6 in both mouse corneas and HCECs.
The nanobody was successfully expressed through microbial system and showed specific high-affinity binding to recombinant dectin 1. Furthermore, it exhibited specific binding to dectin 1 expressed on the surfaces of cells and recognized dectin 1 in mouse corneas. Importantly, it reduced clinical scores of . keratitis in mice compared with a PBS-treatment group. In addition, it decreased mRNA and protein expression of IL1β and IL6 in infected corneas and HCECs stimulated with .
These results suggest that this nanobody can bring about anti-inflammatory effects. This highlights the potential of these nanobodies as innovative therapeutic agents in .
制备一种针对 dectin 1 的纳米抗体，并验证其对角膜炎的特异性和抗炎作用。
采用噬菌体展示技术构建的高质量鲨鱼抗体文库中筛选出纳米抗体。在. 的表达系统中对纳米抗体进行开发。间接 ELISA 用于确定纳米抗体对重组 dectin 1 蛋白的特异性。通过免疫荧光检测纳米抗体在细胞和角膜表面的潜在识别和表达，并进一步验证其对角膜炎的抗炎作用。用感染后，用纳米抗体处理 C57B L/6 小鼠的眼睛。用纳米抗体预处理人角膜上皮细胞（HCEC），然后用孵育。临床评分和裂隙灯照相用于评估小鼠角膜的疾病反应。RT-PCR 和 ELISA 用于评估小鼠角膜和 HCEC 中 IL1β 和 IL6 的 mRNA 和蛋白表达。
通过微生物系统成功表达了纳米抗体，该抗体与重组 dectin 1 具有特异性高亲和力的结合。此外，它与细胞表面表达的 dectin 1 特异性结合，并识别小鼠角膜中的 dectin 1。重要的是，与 PBS 治疗组相比，它降低了. 角膜炎小鼠的临床评分。此外，它还降低了感染角膜和受刺激的 HCEC 中 IL1β 和 IL6 的 mRNA 和蛋白表达。
这些结果表明，该纳米抗体可以产生抗炎作用。这突出了这些纳米抗体作为创新治疗剂在. 的潜力。
