Department of Ophthalmology, The Affiliated Hospital of Qingdao University, Qingdao 266003, China.
College of Food Science and Engineering, Ocean University of China, Qingdao 266404, China.
ACS Infect Dis. 2024 Aug 9;10(8):2991-2998. doi: 10.1021/acsinfecdis.4c00334. Epub 2024 Jul 31.
: to explore the anti-inflammatory effects of a nanobody (Nb) specific to β-glucan on fungal keratitis (FK). : in order to verify the therapeutic and anti-inflammatory efficacy of Nb in FK, the severity of inflammation was assessed with inflammatory scores, hematoxylin-eosin (HE) staining, and myeloperoxidase (MPO) assays. In corneas of mice of FK model and human corneal epithelial cells stimulated by fungal hyphae, real-time reverse transcriptase polymerase chain reaction, Western blot, and enzyme-linked immunosorbent assay were used to detect the expression levels of inflammatory cytokines and pattern recognition receptors (PRRs). In vivo, macrophages and neutrophils infiltration in the cornea stroma was detected by immunofluorescence (IFS) staining. Results: In murine models infected with (), Nb treatment could reduce the inflammatory scores. HE staining and MPO results showed Nb significantly alleviated corneal edema and reduced inflammatory cell infiltration 3 days post-infection. In addition, the expression levels of LOX-1 and Dectin-1 were significantly decreased in the Nb group in vivo. The expression of chemokines CCL2 and CXCL2 also decreased in the Nb group. Compared with the PBS group, the number of macrophages and neutrophils in the Nb group was significantly decreased, which was shown in IFS results. Moreover, Nb attenuated the expression of Dectin-1, LOX-1, and inflammatory mediators, including IL-6 and IL-8 in vitro. Conclusion: our study showed that Nb could alleviate FK by downregulating the expression of PRRs and inflammatory factors as well as reducing the infiltration of macrophages and neutrophils.
: 探索针对β-葡聚糖的纳米抗体（Nb）对真菌性角膜炎（FK）的抗炎作用。: 为了验证 Nb 在 FK 中的治疗和抗炎疗效，通过炎症评分、苏木精-伊红（HE）染色和髓过氧化物酶（MPO）测定评估炎症严重程度。在 FK 模型小鼠和真菌菌丝刺激的人角膜上皮细胞中，实时逆转录聚合酶链反应、Western blot 和酶联免疫吸附测定用于检测炎症细胞因子和模式识别受体（PRRs）的表达水平。在体内，通过免疫荧光（IFS）染色检测角膜基质中巨噬细胞和中性粒细胞的浸润。结果：在感染 的小鼠模型中（），Nb 治疗可降低炎症评分。HE 染色和 MPO 结果表明，Nb 可显著减轻感染后 3 天的角膜水肿和炎症细胞浸润。此外，体内 Nb 组 LOX-1 和 Dectin-1 的表达水平明显降低。趋化因子 CCL2 和 CXCL2 的表达在 Nb 组也降低。与 PBS 组相比，IFS 结果显示 Nb 组的巨噬细胞和中性粒细胞数量明显减少。此外，Nb 可减弱 Dectin-1、LOX-1 和炎症介质（包括 IL-6 和 IL-8）的表达。结论：本研究表明，Nb 通过下调 PRRs 和炎症因子的表达以及减少巨噬细胞和中性粒细胞的浸润，可减轻 FK。
