Role of Pattern Recognition Receptors in the Modulation of Antimicrobial Peptide Expression in the Corneal Epithelial Innate Response to F. solani.
Role of Pattern Recognition Receptors in the Modulation of Antimicrobial Peptide Expression in the Corneal Epithelial Innate Response to F. solani.
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DOI：
10.1167/iovs.16-20658
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发表时间：
2017-05-01
影响因子：
4.4
通讯作者：
McDermott AM
中科院分区：
文献类型：
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作者：
Kolar SS;Baidouri H;McDermott AM
茄病镰刀菌Fusarium solani（F. solani）角膜炎是一种潜在威胁视力的角膜真菌感染。抗微生物肽（AMP），如人β-防御素（hBD）和cathelicidins，免疫系统的重要组成部分，可能对F.索拉尼角膜炎我们研究了模式识别受体（PRRs）、Dectin-1和TLR 2在F.茄素诱导AMP表达的体外调节。将人角膜上皮细胞（HCECs）暴露于热灭活的F。solani或病原相关分子模式（PAMPs）。solani（酵母聚糖或酵母聚糖耗尽）培养6、12或24小时，然后测定AMP mRNA和蛋白质水平。通过使用siRNA敲低（TLR 2和Dectin-1）或化学抑制剂BAY 61-3606（Dectin-1）证实TLR 2和Dectin-1的参与。使用来自F. solani或PAMP处理的HCEC抗F. solani在hBD 2或LL 37中和抗体存在下的表达。我们证实HCEC表达Dectin-1和TLR 2。HCECs在暴露于热灭活的F后表现出AMP hBD 2和cathelicidin LL 37的上调。solani或PAMP。TLR 2和Dectin-1敲低和BAY 61-3606处理降低AMP mRNA上调，证实PRR参与。F. solani或PAMP处理的HCEC显示出对F. solani和hBD 2或LL 37中和抗体显著降低了这种作用，暗示了这些AMP的参与。这些发现表明Dectin-1和TLR 2在调节F.茄尼诱导的角膜上皮细胞AMP表达。
Fusarium solani (F. solani) keratitis is a potentially sight-threatening fungal infection of the cornea. Antimicrobial peptides (AMPs), such as human β-defensins (hBDs) and cathelicidins, essential components of the immune system, likely have a protective role against F. solani keratitis. We examined the role of pattern recognition receptors (PRRs), Dectin-1, and TLR2 in F. solani–induced modulation of AMP expression in vitro. Human corneal epithelial cells (HCECs) were exposed to heat-inactivated F. solani or pathogen-associated molecular patterns (PAMPs) of F. solani (Zymosan or Zymosan Depleted) for 6, 12, or 24 hours following which AMP mRNA and protein levels were determined. Involvement of TLR2 and Dectin-1 was confirmed by using siRNA knock-down (TLR2 and Dectin-1) or chemical inhibitor BAY 61-3606 (Dectin-1). The functional significance of AMP upregulation was tested using culture supernatant from F. solani or PAMP-treated HCECs against F. solani in the presence of hBD2 or LL37 neutralizing antibody. We confirm that HCECs express Dectin-1 and TLR2. HCECs demonstrated upregulation of AMPs hBD2 and cathelicidin LL37 following exposure to heat-inactivated F. solani or PAMPs. TLR2 and Dectin-1 knockdown and BAY 61-3606 treatment decreased AMP mRNA upregulation confirming PRR involvement. The culture supernatant from F. solani or PAMP-treated HCECs showed substantial killing of F. solani and hBD2 or LL37 neutralizing antibody significantly decreased this effect implicating involvement of these AMPs. These findings demonstrate that Dectin-1 and TLR2 have an important role in regulating F. solani-induced AMP expression in corneal epithelial cells.