Optimizing Adjuvants and Needle Free Delivery Methods for Oral HIV Vaccination

在世界范围内，大多数HIV感染是通过粘膜途径（生殖器、口腔或直肠）发生的。的总目标 这项建议是开发一种疫苗接种方法，诱导强烈的艾滋病毒特异性体液和细胞免疫， 生殖器、肠道和口腔粘膜免疫。我们假设，疫苗，这引起强烈的抗艾滋病毒 在这些粘膜部位的免疫力，将防止感染，并迅速清除感染的细胞非常早的网站， 感染和加强保护。口腔内有丰富的淋巴组织，含有抗原呈递细胞，T 细胞和B细胞，并为疫苗的粘膜递送提供了极好的机会。然而，这条路 部分由于唾液中存在蛋白酶， 可以降解疫苗。这对于蛋白质疫苗尤其如此。在这里，我们的目标是针对口腔 使用无针装置（Syrijet）作为口服疫苗递送系统， 在口腔和肠粘膜部位产生强烈的免疫反应。注射器可以将疫苗输送到口腔 因此，粘膜将1）增强疫苗向局部淋巴组织的递送，这将促进DC的摄取 和2）通过防止口服蛋白酶降解来保持疫苗的完整性。来测试我们 假设，我们将使用由CD 40 L佐剂化DNA、MVA和 表达SHIV免疫原的蛋白疫苗。在我们的临床前猕猴研究中，我们发现CD 40 L- 通过IM途径递送的含佐剂DNA/MVA SIV疫苗提供增强的针对直肠内感染的保护 中和抗性SIV挑战。添加以纳米颗粒为佐剂的全身性蛋白加强 DNA/MVA疫苗的包封的TLR 7/8激动剂强烈增强了血清中的HIV特异性IgG 阴道和直肠分泌物中的抗体也增加。在本提案中，在目标1中，我们将首先优化 方法和口服疫苗的位置，并比较不同佐剂的蛋白免疫原。我们将 比较含和不含syrijet的舌下和口腔免疫接种诱导强SHIV-1的效果。 口腔和肠道粘膜组织中的特异性体液和细胞免疫。我们还将测试其他粘膜 佐剂鞭毛蛋白和dmLT用于蛋白免疫。在目标2中，使用最佳递送方法和佐剂 因此，对于我们在目标1中确定的组合，我们将用进化枝C SHIV进行直肠内激发研究。这 这项研究还将侧重于了解保护的免疫相关性。通过实现这些目标，我们 希望已经确定了一种有效的粘膜疫苗接种方案，以增强对HIV的保护。

The majority of HIV infections occur via mucosal routes (genital, oral or rectal) world-wide. The overall goal of this proposal is to develop a vaccination approach that induces strong HIV-specific humoral and cellular immunity in genital, intestinal and oral mucosae. We hypothesize that vaccines, which elicit strong anti-HIV immunity at these mucosal sites, will prevent infection and rapidly clear infected cells very early at the site of infection and enhance protection. Oral cavity is rich in lymphoid tissue containing antigen presenting cells, T cells and B cells, and provides an excellent opportunity for mucosal delivery of vaccines. However, this route of immunization has been under utilized to deliver vaccines in part due to the presence of proteases in saliva that can degrade vaccines. This is particularly true for protein-based vaccines. Here, we aim to target the oral mucosa for immunization using a needle free device (Syrijet) as an oral vaccine delivery system to induce strong immune responses at oral and intestinal mucosal sites. The syrijet can deliver vaccines into the oral mucosa thus will 1) enhance the delivery of vaccine to local lymphoid tissue that will facilitate uptake by DC and 2) will maintain integrity of the vaccine by preventing degradation by oral proteases. To test our hypothesis, we will use a heterologous prime/boost regimen consisting of CD40L-adjuvanted DNA, MVA and protein vaccines expressing SHIV immunogens. In our preclinical macaque studies we showed that CD40L- adjuvanted DNA/MVA SIV vaccines delivered via IM route provide enhanced protection against intrarectal neutralization resistant SIV challenges. The addition of systemic protein boost adjuvanted with nanoparticle encapsulated TLR7/8 agonist to the DNA/MVA vaccine robustly boosted HIV envelope-specific IgG in serum and also increased antibody in vaginal and rectal secretions. In this proposal, in Aim 1, we will first optimize the method and location of oral vaccine delivery and compare different adjuvants for protein immunogen. We will compare sublingual and buccal immunizations delivered with and without syrijet for induction of strong SHIV- specific humoral and cellular immunity in the oral and gut mucosal tissue. We will also test other mucosal adjuvants flagellin and dmLT for protein immunizations. In Aim 2, using the best delivery method and adjuvant combination that we identify in Aim 1, we will conduct an intrarectal challenge study with clade C SHIV. This study will also focus on understanding the immune correlates for protection. By completion of these Aims we hope to have identified an efficient mucosal vaccination regimen for enhancing protection against HIV.