Functional characterization of ferret CCL20 and CCR6 and identification of chemotactic inhibitors.
Functional characterization of ferret CCL20 and CCR6 and identification of chemotactic inhibitors.
复制标题
雪貂 CCL20 和 CCR6 的功能表征以及趋化抑制剂的鉴定。
DOI：
10.1016/j.cyto.2012.12.015
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发表时间：
2013
期刊：
影响因子：
3.8
通讯作者：
Reinhart,ToddA
中科院分区：
文献类型：
--
作者：
Qin,Shulin;Klamar,CynthiaR;FallertJunecko,BethA;Craigo,Jodi;Fuller,DeborahH;Reinhart,ToddA
CCL20是目前已知的唯一的CCR6受体的趋化因子配体，是一种参与正常和病理免疫反应的粘膜趋化因子。虽然有多个物种的CCL20和ccr6序列的核苷酸序列数据，但雪貂的CCL20和ccr6序列尚未确定。为了加深我们对雪貂感染和免疫模型中免疫功能的了解，我们使用RT-PCR技术获得了雪貂CCL20和ccr6的cDNA序列，并对其编码蛋白进行了功能分析。这两个基因的开放阅读框在不同物种之间高度保守，并且大多与犬类序列密切相关。为了进行功能分析，我们构建了表达雪貂CCR6的单细胞克隆，构建了雪貂CCL20/小鼠IgG2a融合蛋白(fCCL20-mIgG2a)，并通过化学方法合成了fCCL20。表达雪貂CCR6的细胞克隆对fCCL20-mIgG2a融合蛋白和人工合成的雪貂CCL20有化学反应。趋化抑制研究发现多酚表没食子儿茶素没食子酸酯和小鼠γ-疱疹病毒68M3蛋白是fCCL20的抑制剂。表面等离子体共振研究表明，EGCG直接与FCCL20结合。这些结果提供了以前未报道的雪貂免疫基因序列的分子特征，并首次鉴定了CCL20的广谱小分子抑制剂，并揭示了CCL20是疱疹病毒M3蛋白的靶标。
CCL20 is currently the only known chemokine ligand for the receptor CCR6, and is a mucosal chemokine involved in normal and pathological immune responses. Although nucleotide sequence data are available for ccl20 and ccr6 sequences from multiple species, the ferret ccl20 and ccr6 sequences have not been determined. To increase our understanding of immune function in ferret models of infection and vaccination, we have used RT-PCR to obtain the ferret ccl20 and ccr6 cDNA sequences and functionally characterize the encoded proteins. The open reading frames of both genes were highly conserved across species and mostly closely related to canine sequences. For functional analyses, single cell clones expressing ferret CCR6 were generated, a ferret CCL20/mouse IgG2afusion protein (fCCL20-mIgG2a) was produced, and fCCL20 was chemically synthesized. Cell clones expressing ferret CCR6 responded chemotactically to fCCL20-mIgG2a fusion protein and synthetic ferret CCL20. Chemotaxis inhibition studies identified the polyphenol epigallocatechin-3-gallate and the murine γ-herpesvirus 68 M3 protein as inhibitors of fCCL20. Surface plasmon resonance studies revealed that EGCG bound directly to fCCL20. These results provide molecular characterization of previously unreported ferret immune gene sequences and for the first time identify a broad-spectrum small molecule inhibitor of CCL20 and reveal CCL20 as a target for the herpesviral M3 protein.
影响因子：
8.8
作者：
M. Fox
通讯作者：
M. Fox