Delivery of antigen to porcine dendritic cells by fusing antigen with porcine dendritic cells targeting peptide.
Delivery of antigen to porcine dendritic cells by fusing antigen with porcine dendritic cells targeting peptide.
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DOI：
10.3389/fimmu.2022.926279
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发表时间：
2022
影响因子：
7.3
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中科院分区：
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树突状细胞（Dendritic cells，DC）是一种专职的抗原提呈细胞，能够识别、捕获和加工抗原。将靶向DC的分子与抗原融合可以有效地提高DC识别和捕获抗原的效率。这种靶向策略可用于疫苗开发，以有效提高DC识别和捕获抗原的效率。本研究确定了与猪DC结合的猪细胞毒性T淋巴细胞相关蛋白4（CTLA 4）的靶向序列。重组罗伊氏乳杆菌（L. reuteri）表达与猪流行性腹泻病毒（PEDV）保护性抗原核心中和表位（COE）融合的CTLA 4 -6aa（LYPPPY）和CTLA 4 - 87 aa，以评价两种靶向基序结合DC上的B7分子的能力。结果表明，CTLA 4 -6aa能与猪DC结合，表达CTLA 4 -6aa的重组乳酸杆菌能更有效地与猪DC结合。此外，DC标志物、toll样受体和细胞因子的表达在6aa-COE/L中显著更高。与用87 aa-COE/L处理的DC相比，reuteri（p<0.01）和表达CTLA 4 -6aa的重组乳杆菌增强了猪DC激活T细胞增殖的能力。我们的蛋白质结构的分析表明，CTLA 4 - 87 aa含有分子内氢键，这可能削弱了猪CTLA 4上的残基和B7上的残基之间的分子间力。结论：表达CTLA 4 -6aa的重组乳酸杆菌能更有效地被猪DC捕获，并具有更强的促进DC成熟和增强T细胞增殖的能力。LYPPPY基序是与猪DC结合的最佳序列。用重组乳杆菌免疫的仔猪显示表达CTLA 4 -6aa的重组乳杆菌诱导显著水平的抗PEDV特异性IgG和伊加抗体应答。本研究可促进DC靶向策略的研究，以提高猪疫苗的有效性。
Dendritic cells (DCs) are professional antigen-presenting cells that can recognize, capture, and process antigens. Fusing molecules targeting DCs with antigens can effectively improve the efficiency with which antigens are recognized and captured by DCs. This targeting strategy can be used for vaccine development to effectively improve the efficiency of antigen recognition and capture by DCs. The targeting sequence of porcine cytotoxic T-lymphocyte associated protein 4 (CTLA4), which binds porcine DCs, was identified in this study. Recombinant Lactobacillus reuteri (L. reuteri) expressing CTLA4-6aa (LYPPPY) and CTLA4-87aa fused to the porcine epidemic diarrhea virus (PEDV) protective antigen core neutralizing epitope (COE) were used to evaluate the ability of the two targeting motifs to bind the B7 molecule on DCs. Our results demonstrate that CTLA4-6aa could bind porcine DCs, and recombinant Lactobacillus expressing the CTLA4-6aa captured by porcine DCs was more efficient than those expressing CTLA4-87aa. In addition, the expression of DC markers, toll-like receptors, and cytokines was significantly higher in the 6aa-COE/L. reuteri-stimulated porcine DCs compared to DCs treated with 87aa-COE/L. reuteri (p<0.01) and recombinant Lactobacillus expressing CTLA4-6aa enhanced the ability of porcine DCs to activate T-cell proliferation. Our analysis of the protein structure revealed that CTLA4-87aa contains intramolecular hydrogen bonds, which may have weakened the intermolecular force between the residues on porcine CTLA4 and that on B7. In conclusion, recombinant Lactobacillus expressing CTLA4-6aa were more efficiently captured by porcine DCs and had a stronger ability to promote DC maturation and enhance T-cell proliferation. The LYPPPY motif is the optimal sequence for binding to porcine DCs. Piglets immunized with recombinant Lactobacillus showed that recombinant Lactobacillus expressing CTLA4-6aa induced significant levels of anti-PEDV-specific IgG and IgA antibody responses. Our study may promote research on DC-targeting strategies to enhance the effectiveness of porcine vaccines.