Screening and Identification of a Chicken Dendritic Cell Binding Peptide by Using a Phage Display Library
Screening and Identification of a Chicken Dendritic Cell Binding Peptide by Using a Phage Display Library
复制标题
DOI：
10.3389/fimmu.2019.01853
复制
发表时间：
2019-08-06
影响因子：
7.3
通讯作者：
Li, Yijing
中科院分区：
文献类型：
--
作者：
Ma, Sunting;Qiao, Xinyuan;Li, Yijing
树突状细胞作为抗原呈递细胞，可以有效地启动适应性免疫反应。虽然dc靶向策略越来越受到关注，但对鸡的相关研究却很少。通过噬菌体展示肽库选择特异性的鸡骨髓dc结合肽，并通过ELISA、流式细胞术、荧光显微镜和激光共聚焦显微镜进行确认。候选肽SPHLHTSSPWER被命名为SP，与传染性法氏囊病病毒（IBDV）结构蛋白和保护性抗原VP2融合。体外实验中，vp2 - sp刺激的DC在4h时的DC标志物（CD80、CD83、CD86、DEC205和MHCII）和一些细胞因子（ifn - γ、IL-12、tnf - α、IL-1 β、IL-6和CXCLi1）的表达显著高于vp2对照肽处理的DC （p < 0.001）。此外，利用鸡源性saerimneri乳杆菌M11 （L. sae M11）制备了一种针对树突状病毒的口服疫苗，将VP2与SP融合，通过口服给药可有效诱导抗ibdv粘膜和体液免疫反应，VP2-SP组的保护效果高于VP2组。因此，L. sae传递的鸡DC靶向IBDV保护抗原VP2对鸡具有有效的免疫保护作用。本研究可促进dc靶向策略的研究，以提高鸡疫苗的有效性。
Dendritic cells (DCs), as antigen-presenting cells, can initiate adaptive immune responses efficiently. Although the DC-targeting strategy has attracted more attention, relevant studies on chicken are rare. Here, specific chicken bone marrow DC-binding peptides were selected using a phage display peptide library and confirmed through ELISA, flow cytometry, fluorescence microscopy, and laser confocal microscopy. The peptide candidate SPHLHTSSPWER, named SP, was fused to the infectious bursal disease virus (IBDV) structural protein and protective antigen VP2. In vitro, the expression of DC markers (CD80, CD83, CD86, DEC205, and MHCII) and some cytokines (IFN-gamma, IL-12, TNF-alpha, IL-1 beta, IL-6, and CXCLi1) by VP2-SP-stimulated DCs was significantly higher than that by DCs treated with the VP2-control peptide at 4h (p < 0.001). In addition, an oral vaccine targeting DCs was generated using chicken-borne Lactobacillus saerimneri M11 (L. sae M11) to deliver VP2 fused with SP. Anti-IBDV mucosal and humoral immune responses were induced efficiently via oral administration, resulting in higher protective efficacy in the VP2-SP group than the VP2 group. Therefore, chicken DC targeting of IBDV protective antigen VP2 delivered by L. sae provides effective immune protection in chicken. Our study may promote research on the DC-targeting strategy to enhance the effectiveness of chicken vaccines.