Liver resident-memory CD8⁺ T cells (T_RM cells) can kill liver-stage Plasmodium-infected cells and prevent malaria, but simple vaccines for generating this important immune population are lacking. Here, we report the development of a fully synthetic self-adjuvanting glycolipid-peptide conjugate vaccine designed to efficiently induce liver T_RM cells. Upon cleavage in vivo, the glycolipid-peptide conjugate vaccine releases an MHC I–restricted peptide epitope (to stimulate Plasmodium-specific CD8⁺ T cells) and an adjuvant component, the NKT cell agonist α-galactosylceramide (α-GalCer). A single dose of this vaccine in mice induced substantial numbers of intrahepatic malaria-specific CD8⁺ T cells expressing canonical markers of liver T_RM cells (CD69, CXCR6, and CD101), and these cells could be further increased in number upon vaccine boosting. We show that modifications to the peptide, such as addition of proteasomal-cleavage sequences or epitope-flanking sequences, or the use of alternative conjugation methods to link the peptide to the glycolipid improved liver T_RM cell generation and led to the development of a vaccine able to induce sterile protection in C57BL/6 mice against Plasmodium berghei sporozoite challenge after a single dose. Furthermore, this vaccine induced endogenous liver T_RM cells that were long-lived (half-life of ~425 days) and were able to maintain >90% sterile protection to day 200. Our findings describe an ideal synthetic vaccine platform for generating large numbers of liver T_RM cells for effective control of liver-stage malaria and, potentially, a variety of other hepatotropic infections.

肝中的常驻CD8 ⁺ T细胞（T _RM细胞）可以杀死肝脏阶段的疟原虫感染的细胞并预防疟疾，但是缺少产生这种重要免疫种群的简单疫苗。在这里，我们报道了一种完全合成的自我佐剂糖脂-肽结合疫苗的开发，该疫苗旨在有效诱导肝T _RM细胞。体内裂解后，糖脂-肽结合疫苗释放MHC I限制的肽表位（以刺激疟原虫特异性CD8 ⁺ T细胞）和佐剂组分NKT细胞激动剂α-半乳糖基神经酰胺（α-GalCer）。在小鼠中单剂这种疫苗可诱导大量肝内疟疾特异性CD8⁺ T细胞表达肝脏T _RM细胞的经典标志物（CD69，CXCR6和CD101），在接种疫苗后这些细胞的数量可能会进一步增加。我们显示出对肽的修饰，例如添加蛋白酶体切割序列或表位侧翼序列，或使用替代缀合方法将肽连接到糖脂上，可改善肝脏T _RM细胞的产生，并导致开发疫苗单次给药后能够诱导C57BL / 6小鼠抵抗伯氏疟原虫子孢子攻击的无菌保护。此外，这种疫苗可诱导内源性肝_TRM寿命长（半衰期约425天）并且能够在200天之前保持> 90％无菌保护的细胞。我们的发现描述了一种理想的合成疫苗平台，该平台可产生大量肝脏T _RM细胞以有效控制肝期疟疾，并可能还有其他各种肝病感染。