CLEC-1 Acts as a Negative Regulator of Dectin-1 Induced Host Inflammatory Response Signature in Aspergillus fumigatus Keratitis
| Autor: | Guoqiang Zhu, Jintao Sun, Limei Wang, Xiaomeng Chen, Chengye Che, Haijing Yan, Ji Eun Lee, Jie Zhang, Leyu Lyu, Hua Yang, Shasha Xue |
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| Rok vydání: | 2021 |
| Předmět: |
0301 basic medicine
Blotting Western Genetic Vectors Biology Real-Time Polymerase Chain Reaction Slit Lamp Microscopy Immunofluorescence Microbiology Keratitis Aspergillus fumigatus Cornea Mice 03 medical and health sciences 0302 clinical medicine Western blot In vivo medicine Animals Aspergillosis Humans Lectins C-Type Fungal keratitis Peroxidase medicine.diagnostic_test Macrophages Membrane Proteins Dependovirus C-type lectin-like receptor-1 (CLEC-1) medicine.disease biology.organism_classification Mice Inbred C57BL Disease Models Animal 030104 developmental biology Real-time polymerase chain reaction Gene Expression Regulation Microscopy Fluorescence Neutrophil Infiltration IL-1β Receptors Mitogen 030220 oncology & carcinogenesis Myeloperoxidase biology.protein Cytokines Female Eye Infections Fungal Dectin-1 |
| Zdroj: | Investigative Ophthalmology & Visual Science |
| ISSN: | 1552-5783 |
| Popis: | Purpose C-type lectin-like receptor-1 (CLEC-1) is a member of the Dectin-1 cluster of pattern recognition receptors (PRRs). It is involved in host immunity, has immunoregulatory function, and supports allograft tolerance. Our study aimed to describe the role of CLEC-1 in response to fungal keratitis, in situ, in vivo, and in vitro. Methods Quantitative polymerase chain reaction (qRT-PCR) and immunofluorescence were used to detect the expression of CLEC-1 in corneas of patients with Aspergillus fumigatus (A. fumigatus) keratitis. In vitro and in vivo experiments were designed in THP-1 macrophages and C57BL/6 mouse models, respectively. The expression of CLEC-1 in corneas of mice model was determined by qRT-PCR, Western blot, and immunofluorescence. CLEC-1 overexpression in mouse corneas was achieved by intrastromal injection of adeno-associated virus (AAV) vectors. Disease response was evaluated by slit-lamp photography, clinical score, and colony forming unit (CFU). Bioluminescence imaging system image acquisition, myeloperoxidase (MPO) assays, immunofluorescence staining, qRT-PCR, and Western blot were used to investigate the role of CLEC-1. To further define the role of CLEC-1, we used lentivirus vectors to overexpress CLEC-1 or/and Dectin-1 in THP-1 macrophages. Results The expression of CLEC-1 was increased in corneas of patients with A. fumigatus keratitis. In corneas of mice from the A. fumigatus keratitis model, the expression of CLEC-1 was decreased in the acute inflammatory stage and increased during convalescence. Following Natamycin treatment, CLEC-1 was upregulated in A. fumigatus keratitis mice. Compared with normal C57BL/6 mice, overexpression of CLEC-1 converted the characteristic susceptible response to resistance, as demonstrated by slit-lamp photography and clinical score. In vivo studies revealed decreased MPO levels and neutrophils recruitment and higher fungal load after the upregulation of CLEC-1. Compared with control corneas, CLEC-1 overexpression impaired corneal pro-inflammatory cytokine IL-1β production. Conclusions These findings demonstrate that CLEC-1 may act as a negative regulator of Dectin-1 induced host inflammatory response via suppressing neutrophils recruitment and production of pro-inflammatory cytokine IL-1β production in response to A. fumigatus keratitis. |
| Databáze: | OpenAIRE |
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