Mechanisms of DDO Adjuvancy

总结 能够诱导持久的1型细胞免疫的佐剂，包括Th1 CD4 + T细胞和 细胞毒性CD8 + T细胞，被高度寻求用于开发针对细胞内病原体的疫苗 能逃避抗体我们已经确定了一个强大的新一类佐剂，eliminating保护1型偏向 应答这些佐剂是来自缺陷病毒基因组（DDO）的合成RNA寡核苷酸。 并通过接合细胞RIG-I样受体（RLR）触发强烈的免疫应答。DDO包含唯一的 免疫刺激基序，我们确定为必要的能力，触发RLR信号。在小鼠中，DDO 诱导I型IFN和其它细胞因子的局部表达，并促进DC在 引流淋巴结此外，DDO促进I型IFN依赖性IgG2b/c偏向性抗体应答 能够保护小鼠免受致死性病毒攻击并诱导产生IFN γ的抗原特异性CD4+和CD8 + T细胞 细胞此外，DDO与基于角鲨烯的佐剂AddaVax协同作用， 对用AddaVax + DDO佐剂的疫苗的免疫偏倚。这些数据支持我们的中心假设， DDO代表了一类新的佐剂，其刺激RLR/I型IFN信号传导轴以驱动免疫应答。 最佳的长寿1型体液和细胞免疫。 该提案中的实验使用了洛佩斯和 Scott实验室评估DDO诱导的T细胞应答的质量、寿命和保护能力 在疫苗接种过程中，并表征负责指导免疫的特定分子和细胞机制， DDO给药后的1型免疫应答。具体来说，在目标1中，我们将使用我们的能力来跟踪 体内DDO，以及一系列报告基因和转基因小鼠，以确定DDO的早期相互作用 与免疫系统的细胞，特别是DC，并确定潜在的关键目标， -1型偏向反应。在目标2中，我们将评估辅助性T细胞1、细胞毒性T细胞和细胞因子的发育和质量。 细胞、T滤泡辅助细胞和组织驻留T细胞对单独用DDO佐剂的疫苗的应答 或与AddaVax组合，并评估I型IFN在建立这些应答中的作用。在目标3中， 我们将测试DDO诱导CD4+介导的抗细胞内原生动物寄生虫的能力 利什曼原虫，因为对这种寄生虫的保护依赖于CD4 + Th1细胞，而不依赖于 抗体，我们将使用模型直接评估DDO诱导保护性T细胞应答的能力。 雪貂的流感疫苗

SUMMARY Adjuvants able to induce long lasting type-1 cellular immunity, including both Th1 CD4+ T cells and cytotoxic CD8+ T cells, are highly sought out for the development of vaccines against intracellular pathogens that evade antibodies. We have identified a powerful new class of adjuvant that elicits protective type-1 biased responses. These adjuvants are synthetic RNA oligonucleotides derived from defective viral genomes (DDOs) and trigger strong immune responses by engaging cellular RIG-I-like receptors (RLRs). DDOs contain a unique immunostimulatory motif that we identified as essential for their ability to trigger RLR signaling. In mice, DDOs induce localized expression of type I IFNs and other cytokines and promote accumulation of DCs in the draining lymph node. In addition, DDOs promote a type I IFN-dependent IgG2b/c-biased antibody response able to protect mice from lethal virus challenge and induce IFNγ-producing antigen-specific CD4+ and CD8+ T cells. Moreover, DDOs synergize with the squalene-based adjuvant AddaVax, providing strong type-1 immunity bias to vaccines adjuvanted with AddaVax+DDOs. These data support our central hypothesis that DDOs represent a new class of adjuvants that stimulate the RLR/type I IFN signaling axis to drive optimal long-lived type-1 humoral and cellular immunity. Experiments in this proposal use the combined expertise and unique set of tools of the Lopez and Scott laboratories to assess the quality, longevity, and protective capacity of DDO-induced T cell responses during vaccination, and to characterize specific molecular and cellular mechanisms responsible for directing the type-1 immune response after DDO administration. Specifically, in Aim 1 we will use our ability to track DDOs in vivo, together with a series of reporter and transgenic mice, to identify the early interactions of DDOs with cells of the immune system, in particular DCs, and to identify potential key targets for the development of type -1 biased responses. In Aim 2, we will assess the development and quality of T helper 1 cells, cytotoxic T cells, T follicular helper cells, and tissue resident T cells in response to a vaccine adjuvanted with DDOs alone or in combination with AddaVax, and assess the role of type I IFNs in establishing these responses. In Aim 3, we will test the ability of DDOs to induce CD4+-mediated protection against the intracellular protozoan parasite Leishmania, since protection against this parasite is dependent upon CD4+ Th1 cells and is independent of antibodies, and we will directly assess the ability of DDOs to induce protective T cell responses using a model influenza vaccine in ferrets.