TNFSF APC activators for HIV Vaccines

描述（由申请人提供）：需要活化的抗原呈递细胞（apc）来诱导强烈和持久的免疫反应。对于疫苗，apc通常由疫苗制剂中的化学成分或微生物成分激活。相比之下，正常的免疫应答依赖于apc的内源性激活剂，主要是CD40配体（CD40L，也称为CD154或TNFSF5）。虽然许多研究已经表明CD40L对疫苗反应有多重要，但这种蛋白质的理想疫苗配方尚未确定。CD40L的表达质粒导致非常强的免疫反应，但可溶性三聚体形式的蛋白活性较低。这项R03小型研究资助将测试以三种不同形式传递CD40L的可行性：作为膜CD40L，作为可溶性CD40L三聚体，以及作为与表面活性剂蛋白d融合蛋白形成的新型12链四臂结构。此外，TNF超家族的其他成员已被提出用于增强免疫应答，但尚未在HIV疫苗环境中进行测试。其中两种TNFSF配体也将被研究：RANKL （TRANCE, TNFSF11）和CD27L （CD70, TNFSF7）。每个TNFSF配体将与HIV gp120 Env或p24 gag的质粒DNA结构一起施用于小鼠。疫苗反应将通过抗体形成、淋巴细胞增殖反应、细胞因子产生和细胞毒性测定来测定。此外，CD40L构建体将与分枝杆菌hsp70和HIV p24 Gag制成的融合蛋白结合，以确定CD40L与现有疫苗结合时是否具有加性效应。在完成这些可行性研究后，是否应该将这些概念推进到针对艾滋病毒和其他病原体的更广泛的疫苗研究中将会很清楚。

DESCRIPTION (provided by applicant): Activated antigen-presenting cells (APCs) are required in order to induce a strong and long-lasting immune response. For vaccines, APCs are usually activated by either chemical or microbial components in the vaccine formulation. In contrast, normal immune responses rely upon endogenous activators of APCs, mainly CD40 ligand (CD40L, also called CD154 or TNFSF5). While numerous studies have shown how important CD40L is for vaccine responses, an ideal vaccine formulation of this protein has not been determined. Expression plasmids for CD40L result in a very strong immune response, but a soluble trimeric form of the protein is less active. This R03 Small Research Grant will test the feasibility of delivering CD40L in three different forms: as membrane CD40L, as a soluble CD40L trimer, and as a novel 12-chain, four-armed structure formed as a fusion protein with the body of surfactant protein D. Additionally, other members of the TNF superfamily of ligands have been proposed to augment immune responses but have not been tested in a HIV vaccine setting. Two of these TNFSF ligands will also be studied: RANKL (TRANCE, TNFSF11) and CD27L (CD70, TNFSF7). Each TNFSF ligand will be co-administered to mice along with a plasmid DNA construct for either HIV gp120 Env or p24 gag. Vaccine responses will be measured using assays for antibody formation, lymphoproliferative responses, cytokine production, and cytotoxity. Additionally, CD40L constructs will be combined with a fusion protein made from mycobacterial hsp70 and HIV p24 Gag to determine if CD40L has an additive effect when combined with an existing vaccine. Upon the completion of these feasibility studies, it will be clear if these concepts should be advanced into more extensive vaccine studies both for HIV and other pathogens.