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噬菌体展示技术针对角膜上皮细胞筛选出具有生物活性的抑制曲霉菌黏附角膜的短肽。

Phage display against corneal epithelial cells produced bioactive peptides that inhibit Aspergillus adhesion to the corneas.

机构信息

Shandong Provincial Key Laboratory of Ophthalmology, Shandong Eye Institute, Shandong Academy of Medical Sciences, Qingdao, China.

出版信息

PLoS One. 2012;7(3):e33578. doi: 10.1371/journal.pone.0033578. Epub 2012 Mar 12.

DOI:10.1371/journal.pone.0033578
PMID:22428072
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC3299800/
Abstract

Dissection of host-pathogen interactions is important for both understanding the pathogenesis of infectious diseases and developing therapeutics for the infectious diseases like various infectious keratitis. To enhance the knowledge about pathogenesis infectious keratitis, a random 12-mer peptide phage display library was screened against cultured human corneal epithelial cells (HCEC). Fourteen sequences were obtained and BLASTp analysis showed that most of their homologue counterparts in GenBank were for defined or putative proteins in various pathogens. Based on known or predicted functions of the homologue proteins, ten synthetic peptides (Pc-A to Pc-J) were measured for their affinity to bind cells and their potential efficacy to interfere with pathogen adhesion to the cells. Besides binding to HCEC, most of them also bound to human corneal stromal cells and umbilical endothelial cells to different extents. When added to HCEC culture, the peptides induced expression of MyD88 and IL-17 in HCEC, and the stimulated cell culture medium showed fungicidal potency to various extents. While peptides Pc-C and Pc-E inhibited Aspergillus fumigatus (A.f) adhesion to HCEC in a dose-dependent manner, the similar inhibition ability of peptides Pc-A and Pc-B required presence of their homologue ligand Alb1p on A.f. When utilized in an eyeball organ culture model and an in vivo A.f keratitis model established in mouse, Pc-C and Pc-E inhibited fungal adhesion to corneas, hence decreased corneal disruption caused by inflammatory infiltration. Affinity pull-down of HCEC membrane proteins with peptide Pc-C revealed several molecules as potential receptors for this peptide. In conclusion, besides proving that phage display-selected peptides could be utilized to interfere with adhesion of pathogens to host cells, hence could be exploited for managing infectious diseases including infectious keratitis, we also proposed that the phage display technique and the resultant peptides could be used to explore host-pathogen interactions at molecular levels.

摘要

宿主-病原体相互作用的解析对于理解传染病的发病机制以及开发治疗各种感染性角膜炎等传染病的疗法都很重要。为了增强对感染性角膜炎发病机制的认识，针对培养的人角膜上皮细胞（HCEC）筛选了随机的 12 肽噬菌体展示文库。获得了 14 个序列，BLASTp 分析表明，它们在 GenBank 中的同源物大多数是各种病原体中已定义或推测的蛋白质。基于同源蛋白的已知或预测功能，测量了 10 个合成肽（Pc-A 至 Pc-J）与细胞结合的亲和力及其潜在的干扰病原体与细胞黏附的功效。除了与 HCEC 结合外，它们中的大多数还不同程度地与人角膜基质细胞和脐静脉内皮细胞结合。当添加到 HCEC 培养物中时，这些肽诱导 HCEC 中 MyD88 和 IL-17 的表达，并且刺激的细胞培养液具有不同程度的杀菌效力。虽然肽 Pc-C 和 Pc-E 以剂量依赖的方式抑制烟曲霉（A.f）与 HCEC 的黏附，但肽 Pc-A 和 Pc-B 的类似抑制能力需要其同源配体 Alb1p 在 A.f 上的存在。当在眼球器官培养模型和在小鼠中建立的体内 A.f 角膜炎模型中使用时，Pc-C 和 Pc-E 抑制了真菌对角膜的黏附，从而减少了炎症浸润引起的角膜破坏。用肽 Pc-C 亲和拉下 HCEC 膜蛋白揭示了几种作为该肽潜在受体的分子。总之，除了证明噬菌体展示选择的肽可用于干扰病原体与宿主细胞的黏附，从而可用于治疗包括感染性角膜炎在内的传染病外，我们还提出噬菌体展示技术和由此产生的肽可用于在分子水平上探索宿主-病原体相互作用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3f67/3299800/87614bb63ca4/pone.0033578.g008.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3f67/3299800/7e95852941cb/pone.0033578.g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3f67/3299800/e20bbb6996fc/pone.0033578.g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3f67/3299800/ad8d1b824d8c/pone.0033578.g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3f67/3299800/ae20cb4eee7f/pone.0033578.g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3f67/3299800/4979ea83664f/pone.0033578.g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3f67/3299800/895ae0f35793/pone.0033578.g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3f67/3299800/b6ddd0cdc198/pone.0033578.g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3f67/3299800/87614bb63ca4/pone.0033578.g008.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3f67/3299800/7e95852941cb/pone.0033578.g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3f67/3299800/e20bbb6996fc/pone.0033578.g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3f67/3299800/ad8d1b824d8c/pone.0033578.g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3f67/3299800/ae20cb4eee7f/pone.0033578.g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3f67/3299800/4979ea83664f/pone.0033578.g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3f67/3299800/895ae0f35793/pone.0033578.g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3f67/3299800/b6ddd0cdc198/pone.0033578.g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3f67/3299800/87614bb63ca4/pone.0033578.g008.jpg

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[4]
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[5]
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[6]
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[7]
Peptide therapeutics for treating ocular surface infections.

J Ocul Pharmacol Ther. 2014-11

本文引用的文献

[1]
3D quantitative imaging of unprocessed live tissue reveals epithelial defense against bacterial adhesion and subsequent traversal requires MyD88.

PLoS One. 2011-8-25

[2]
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J Immunol. 2011-7-15

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