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Screening and Identification of a Chicken Dendritic Cell Binding Peptide by Using a Phage Display Library

机译:噬菌体展示文库筛选和鉴定鸡树突状细胞结合肽

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摘要

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-γ, IL-12, TNF-α, IL-1β, IL-6, and CXCLi1) by VP2-SP-stimulated DCs was significantly higher than that by DCs treated with the VP2-control peptide at 4 h (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.
机译:树突状细胞(DC)作为抗原呈递细胞,可以有效地启动适应性免疫反应。尽管DC靶向策略引起了更多关注,但有关鸡肉的相关研究却很少。在这里,使用噬菌体展示肽库选择特定的鸡骨髓DC结合肽,并通过ELISA,流式细胞仪,荧光显微镜和激光共聚焦显微镜进行确认。候选肽SPHLHTSSPWER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。在体外,VP2-表达DC标记(CD80,CD83,CD86,DEC205和MHCII)和某些细胞因子(IFN-γ,IL-12,TNF-α,IL-1β,IL-6和CXCLi1)。 SP刺激的DC显着高于用VP2-对照肽处理4 h的DC(p <0.001)。另外,使用鸡源的啤酒杆菌M11(L. sae M11)产生了靶向DC的口服疫苗,以递送与SP融合的VP2。通过口服给药可有效诱导抗IBDV黏膜和体液免疫反应,与VP2组相比,VP2-SP组的保护作用更高。因此,由赛氏乳杆菌递送的针对IBDV保护性抗原VP2的鸡DC靶向在鸡中提供了有效的免疫保护。我们的研究可能会促进对DC靶向策略的研究,以提高鸡疫苗的有效性。

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