Combined TNFSF and TLR stimulation of HIV vaccines

描述（由申请人提供）：开发有效的HIV感染疫苗的一个障碍是需要一种导致强烈免疫应答的疫苗接种方法。DNA疫苗具有潜力，但它们通常是较差的免疫原，除非添加佐剂质粒。实验表明，质粒DNA的一个新的可溶性，多聚体形式的CD 40 L是一种有效的佐剂的DNA疫苗在小鼠模型中，它将是重要的测试其他免疫刺激成员的TNF超家族（TNFSF）。 组合Toll样受体（TLR）和CD 40刺激是高度协同的，并且将使用上述新形式的CD 40 L进行测试。此外，用PLGA/PBAE共聚物微囊化DNA疫苗有助于将DNA携带至树突状细胞，并且还上调CD 40，表明其可能与作为佐剂的CD 40 L协同作用。因此，本项目有以下目的：目的1：检测OX 40 L、RANKL、CD 27 L和LIGHT质粒对小鼠HIV DNA疫苗接种的个体效应。 目的2：研究CD 40配体与细菌脂肽TLR 2激动剂联合应用作为DNA疫苗佐剂的效果。目的3：探讨PLGA/PBAE微囊化DNA与CD 40配体协同增强HIV DNA疫苗免疫应答的作用。目的4：制备猕猴版本的最佳TNFSF构建体，用于推进SIV/猕猴模型。该R21提案的创新方面是：它使用了一种有前途的新的CD 40 L和其他TNFSF递送平台;研究了CD 40 L加BLP作为DNA疫苗佐剂;并将CD 40 L与PLGA/PBAE微囊化结合在HIV DNA疫苗中。总之，这些研究将优化HIV DNA疫苗的强大佐剂平台，该平台对于仅DNA疫苗接种或DNA初免/病毒载体加强方案具有巨大潜力。此外，这种DNA疫苗方法可能被证明普遍适用于新出现的感染和其他需要有效疫苗的疾病。

DESCRIPTION (provided by applicant): One impediment to developing an effective vaccine for HIV infection is the requirement for a vaccination method that leads to strong immune responses. DNA vaccines have potential, but they are often poor immunogens unless an adjuvant plasmid is added. Experiments are described that show that plasmid DNA for a new soluble, multimeric form of CD40L is an effective adjuvant for DNA vaccines in mouse models, and it will be important to test other immunostimulatory members of the TNF superfamily (TNFSF). Combining Toll-like Receptor (TLR) and CD40 stimulation is highly synergistic, and will be tested using the novel form of CD40L above. Additionally, microencapsulation of DNA vaccines with PLGA/PBAE copolymer helps to carry the DNA to dendritic cells and also upregulates CD40, indicating that it might synergize with CD40L as an adjuvant. Consequently, this project has the following aims: Aim 1: To test the individual effects of plasmids for OX40L, RANKL, CD27L, and LIGHT on HIV DNA vaccination in mice. Aim 2: To test combinations of CD40L and bacterial lipopeptide (BLP) TLR2 agonists as DNA vaccine adjuvants. Aim 3: To determine if DNA microencapsulation with PLGA/PBAE synergizes with CD40L to enhance the immune response to an HIV DNA vaccine. Aim 4: To prepare macaque versions of the best TNFSF constructs for advancement to the SIV/macaque model. The innovative aspects of this R21 proposal are: its use of a promising new delivery platform for CD40L and other TNFSFs; studies of CD40L plus BLPs as DNA vaccine adjuvants; and combining CD40L with PLGA/PBAE microencapsulation in an HIV DNA vaccine. In sum, these studies will optimize a powerful adjuvant platform for HIV DNA vaccines that has great potential for DNA-only vaccination or DNA prime/viral vector boost protocols. In addition, this DNA vaccine approach may prove generally applicable for emerging infections and other diseases for which a potent vaccine is needed.