TNFSF APC activators for HIV Vaccines

描述（由申请方提供）：需要活化的抗原呈递细胞（APC）以诱导强烈和持久的免疫应答。 对于疫苗，APC通常被疫苗制剂中的化学或微生物组分激活。 相反，正常的免疫应答依赖于APC的内源性活化剂，主要是CD40配体（CD40L，也称为CD154或TNFSF 5）。 虽然许多研究已经表明CD40 L对于疫苗应答是多么重要，但这种蛋白质的理想疫苗制剂尚未确定。 CD40L的表达质粒导致非常强的免疫应答，但可溶性三聚体形式的蛋白质活性较低。 这项R03小型研究基金将测试以三种不同形式提供CD40 L的可行性：作为膜CD40 L，作为可溶性CD40 L三聚体，以及作为一种新的12链，四臂结构与表面活性蛋白D的融合蛋白形成。 此外，TNF超家族配体的其他成员已被提议增强免疫应答，但尚未在HIV疫苗环境中进行测试。 还将研究其中两种TNFSF配体：RANKL（TRANCE，TNFSF 11）和CD 27 L（CD 70，TNFSF 7）。 将每种TNFSF配体与HIV gp120 Env或p24 gag的质粒DNA构建体一起沿着给予小鼠。 将使用抗体形成、淋巴增殖反应、细胞因子产生和细胞毒性测定法测量疫苗反应。 此外，CD40 L构建体将与由分枝杆菌hsp70和HIV p24 Gag制成的融合蛋白组合，以确定CD40 L与现有疫苗组合时是否具有累加效应。 在完成这些可行性研究后，将明确这些概念是否应推进到更广泛的艾滋病毒和其他病原体疫苗研究中。

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.