优选的抗原递送模式伴随着足够的佐剂有利于疫苗效率。线粒体具有原核特征，含有多种损伤相关分子模式（DAMP），很容易被吞噬细胞摄取，同时激活先天免疫。在当前的研究中，我们建立了一个线粒体工程平台，用于生成富含抗原的线粒体作为癌症疫苗。以卵清蛋白（OVA）和酪氨酸酶相关蛋白2（TRP2）为模型抗原，合成具有线粒体定位信号肽的融合蛋白。然后使用慢病毒感染系统生产含有 OVA 或 TRP2 的线粒体疫苗。提取含有 OVA 和 TRP2 的工程线粒体（OVA-MITO 和 TRP2-MITO）并作为潜在的癌症疫苗进行评估。令人印象深刻的是，工程线粒体疫苗在小鼠肿瘤模型中用作预防性和治疗性疫苗时表现出有效的抗肿瘤作用。从机制上讲，OVA-MITO 和 TRP2-MITO 有效招募并激活树突状细胞 (DC)，并诱导肿瘤特异性细胞介导的免疫。此外，线粒体疫苗对DC的激活关键涉及TLR2途径及其脂质激动剂，即源自线粒体膜的心磷脂。结果表明，工程化线粒体本身就是精心编排的载体，充满了用于抗原递送的免疫刺激物，可以更好地靶向局部树突状细胞并发挥强大的适应性细胞免疫。这项概念验证研究建立了一个通用的疫苗构建平台，其中包含带有针对癌症和其他疾病的可变抗原的工程线粒体。© 2024。作者。

The preferable antigen delivery profile accompanied by sufficient adjuvants favors vaccine efficiency. Mitochondria, which feature prokaryotic characteristics and contain various damage-associated molecular patterns (DAMPs), are easily taken up by phagocytes and simultaneously activate innate immunity. In the current study, we established a mitochondria engineering platform for generating antigen-enriched mitochondria as cancer vaccine. Ovalbumin (OVA) and tyrosinase-related protein 2 (TRP2) were used as model antigens to synthesize fusion proteins with mitochondria-localized signal peptides. The lentiviral infection system was then employed to produce mitochondrial vaccines containing either OVA or TRP2. Engineered OVA- and TRP2-containing mitochondria (OVA-MITO and TRP2-MITO) were extracted and evaluated as potential cancer vaccines. Impressively, the engineered mitochondria vaccine demonstrated efficient antitumor effects when used as both prophylactic and therapeutic vaccines in murine tumor models. Mechanistically, OVA-MITO and TRP2-MITO potently recruited and activated dendritic cells (DCs) and induced a tumor-specific cell-mediated immunity. Moreover, DC activation by mitochondria vaccine critically involves TLR2 pathway and its lipid agonist, namely, cardiolipin derived from the mitochondrial membrane. The results demonstrated that engineered mitochondria are natively well-orchestrated carriers full of immune stimulants for antigen delivery, which could preferably target local dendritic cells and exert strong adaptive cellular immunity. This proof-of-concept study established a universal platform for vaccine construction with engineered mitochondria bearing alterable antigens for cancers as well as other diseases.© 2024. The Author(s).