Hyun Park Su, Kim Yun Hye, Lee Hyeon Jin, Han Jeong Moo, Seo Byoung-Joo, Park Gyeong-Seo, Kim Chonghan, Ryu Young Bae, Kim Woo Sik
Biological Resource Center, Korea Research Institute of Bioscience and Biotechnology, Jeongeup, Korea.
Functional Biomaterial Research Center, Korea Research Institute of Bioscience and Biotechnology, Jeongeup, Republic of Korea.
Virulence. 2025 Dec;16(1):2453818. doi: 10.1080/21505594.2025.2453818. Epub 2025 Jan 20.
(APP) is a significant pathogen in the swine industry, leading to substantial economic losses and highlighting the need for effective vaccines. This study evaluates the potential of APP-derived extracellular vesicles (APP-EVs) as a vaccine candidate compared to the commercial Coglapix vaccine. APP-EVs, isolated using tangential flow filtration (TFF) and cushioned ultracentrifugation, exhibited an average size of 105 nm and a zeta potential of -17.4 mV. These EVs demonstrated stability under external stressors, such as pH changes and enzymatic exposure and were found to contain 86 major metabolites. Additionally, APP-EVs induced dendritic cell (DC) maturation in a Toll-like receptor 4 (TLR4)-dependent manner without cytotoxicity. APP-EVs predominantly elicited Th1-mediated IgG responses in immunized mice without significant liver and kidney toxicity. Contrarily, unlike Coglapix, which induced stronger Th2-mediated responses and notable toxicity. In addition, APP-EVs triggered APP-specific Th1, Th17, and cytotoxic T lymphocyte (CTL) responses and promoted the activation of multifunctional T-cells. Notably, APP-EV immunization enhanced macrophage phagocytosis and improved survival rates in mice challenged with APP infection compared to those treated with Coglapix. These findings suggest that APP-EVs are promising vaccine candidates, capable of inducing potent APP-specific T-cell responses, particularly Th1, Th17, CTL, and multifunctional T-cells, thereby enhancing the protective immune response against APP infection.
副猪嗜血杆菌（APP）是养猪业中的一种重要病原体，会导致巨大的经济损失，并凸显了有效疫苗的必要性。本研究评估了与市售Coglapix疫苗相比，源自APP的细胞外囊泡（APP-EVs）作为候选疫苗的潜力。使用切向流过滤（TFF）和缓冲超速离心法分离得到的APP-EVs，平均大小为105 nm，zeta电位为-17.4 mV。这些细胞外囊泡在外部应激源（如pH变化和酶暴露）下表现出稳定性，并且发现含有86种主要代谢物。此外，APP-EVs以Toll样受体4（TLR4）依赖的方式诱导树突状细胞（DC）成熟，且无细胞毒性。APP-EVs在免疫小鼠中主要引发Th1介导的IgG反应，且无明显的肝肾毒性。相反，与诱导更强Th2介导反应和明显毒性的Coglapix不同。此外，APP-EVs触发了APP特异性的Th1、Th17和细胞毒性T淋巴细胞（CTL）反应，并促进了多功能T细胞的激活。值得注意的是，与用Coglapix治疗的小鼠相比，APP-EV免疫增强了巨噬细胞吞噬作用，并提高了感染APP的小鼠的存活率。这些发现表明，APP-EVs是有前景的候选疫苗，能够诱导有效的APP特异性T细胞反应，特别是Th1、Th17、CTL和多功能T细胞，从而增强针对APP感染的保护性免疫反应。
