抗病毒和抗肿瘤疫苗旨在诱导细胞毒性CD8+T细胞(CTL)和抗体。保守蛋白抗原,如人类免疫缺陷病毒的p24,代表激发CTL的有希望的成分,然而,免疫原性欠佳,如果配制成重组蛋白。为了增强免疫原性和CTL反应,重组蛋白可靶向树突状细胞(DC)以在MHCI上交叉呈递,其中甘露糖受体和/或其他凝集素受体可发挥重要作用。这里,我们构建了基于脂质体载体的疫苗,该疫苗由重组p24抗原通过金属螯合键结合到纳米脂质体表面,表面甘露聚糖通过氨氧基连接偶联。通过动态光散射分析生成的甘露糖基化蛋白聚糖体,等温滴定,和电子显微镜。使用鼠DC线MutuDC和鼠骨髓来源的DC(BMDC),我们评估了它们的免疫原性和免疫调节活性。我们显示p24甘露糖基化的蛋白酶体激活DC,以增强MutuDC和BMDC上的MHCI,MHCII和CD40,CD80和CD86表面表达。p24甘露糖基化脂质体被MutuDC内化,p24在1至3小时内进行细胞内定位。金属螯合和氨氧基连接的组合可同时用于生成纳米脂质体佐剂化的基于重组蛋白的疫苗,该疫苗通用于与抗体和CTL激发相关的重组抗原的组合。
Anti-viral and anti-tumor vaccines aim to induce cytotoxic CD8+ T cells (CTL) and antibodies. Conserved protein antigens, such as p24 from human immunodeficiency virus, represent promising component for elicitation CTLs, nevertheless with suboptimal immunogenicity, if formulated as recombinant protein. To enhance immunogenicity and CTL response, recombinant proteins may be targeted to dendritic cells (DC) for cross presentation on MHCI, where mannose receptor and/or other lectin receptors could play an important role. Here, we constructed liposomal carrier-based vaccine composed of recombinant p24 antigen bound by metallochelating linkage onto surface of nanoliposomes with surface mannans coupled by aminooxy ligation. Generated mannosylated proteonanoliposomes were analyzed by dynamic light scattering, isothermal titration, and electron microscopy. Using murine DC line MutuDC and murine bone marrow derived DC (BMDC) we evaluated their immunogenicity and immunomodulatory activity. We show that p24 mannosylated proteonanoliposomes activate DC for enhanced MHCI, MHCII and CD40, CD80, and CD86 surface expression both on MutuDC and BMDC. p24 mannosylated liposomes were internalized by MutuDC with p24 intracellular localization within 1 to 3 h. The combination of metallochelating and aminooxy ligation could be used simultaneously to generate nanoliposomal adjuvanted recombinant protein-based vaccines versatile for combination of recombinant antigens relevant for antibody and CTL elicitation.