2. Academic Validation
  3. CXCL16 exacerbates Pseudomonas aeruginosa keratitis by promoting neutrophil activation

CXCL16 exacerbates Pseudomonas aeruginosa keratitis by promoting neutrophil activation

  • Int Immunopharmacol. 2023 Dec 27:127:111375. doi: 10.1016/j.intimp.2023.111375.
Jiayin Wu 1,  Wentao Wang 2,  Fengjiao Yuan 3,  Juan Zheng 3,  Weihua Zhang 4,  Hui Guo 5,  Leyi Wang 5,  Chenyang Dai 5,  Fang Han 5,  Xinyi Wu 5,  Jianlu Gao 6
Affiliations

Affiliations

  • 1 Department of Ophthalmology, Qilu Hospital of Shandong University, Jinan, Shandong 250012, China; Department of Ophthalmology, Liaocheng People's Hospital, Shandong University, Liaocheng, Shandong 252000, China.
  • 2 Department of Surgery, Liaocheng Tumor Hospital, Liaocheng, Shandong 252000, China.
  • 3 Joint Laboratory for Translational Medicine Research, Liaocheng People's Hospital, Shandong University, Liaocheng, Shandong 252000, China.
  • 4 Department of Ophthalmology, Liaocheng People's Hospital, Shandong University, Liaocheng, Shandong 252000, China.
  • 5 Department of Ophthalmology, Qilu Hospital of Shandong University, Jinan, Shandong 250012, China.
  • 6 Department of Ophthalmology, Liaocheng People's Hospital, Shandong University, Liaocheng, Shandong 252000, China. Electronic address: [email protected].
PMID: 38154213 DOI: 10.1016/j.intimp.2023.111375
Abstract

Pseudomonas aeruginosa (PA) keratitis is a major cause of blindness characterized by corneal inflammation. In a murine model of PA keratitis, we assessed the detrimental effects of CXC chemokine ligand 16 (CXCL16). Quantitative PCR (qPCR), western blotting (WB) and immunofluorescence were used to measure the expression and localization of CXCL16 and its receptor, CXC Chemokine Receptor 6 (CXCR6). Clinical scores, plate counting, and hematoxylin-eosin staining were used to assess Infection severity and its exacerbation by CXCL16. Immunofluorescence, myeloperoxidase assays, and flow cytometry were used to detect neutrophil activity and colocalization with CXCR6. WB and immunofluorescence were used to measure levels of Reactive Oxygen Species (ROS) and Matrix Metalloproteinases (MMPs). These methods also were used to measure the activation of downstream NF-κB signaling and its positive feedback on CXCL16 expression. ELISA, flow cytometry, and qPCR were used to measure the expression of CXCL2 and T helper 17 (Th17) cell-related genes. CXCL16 and CXCR6 expression was increased in infected corneas. Topical application of CXCL16 exacerbated keratitis by increasing corneal Bacterial load and promoting neutrophil infiltration, whereas neutralizing antibody against CXCL16 had the opposite effect. CXCL16 also increased ROS and MMP levels. This neutrophil activation may be caused by its positive feedback with the NF-κB pathway and the upregulation of CXCL2 and Th17 cell related-genes. These data suggest that CXCL16 is an attractive therapeutic target for PA keratitis.

Keywords

CXCL16; CXCR6; NF-κB; Neutrophil activation; Pseudomonas aeruginosa (PA) keratitis.

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