Optimizing Adjuvants and Needle Free Delivery Methods for Oral HIV Vaccination

全世界大多数艾滋病毒感染是通过粘膜途径(生殖器、口腔或直肠)发生的。的总目标是 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 生殖器、肠道和口腔粘膜的免疫。我们假设，能够产生强大的抗艾滋病毒的疫苗 这些粘膜部位的免疫将防止感染，并在感染部位很早就迅速清除受感染的细胞 感染，加强防护。口腔富含含有抗原提呈细胞的淋巴组织，T细胞 细胞和B细胞，为疫苗的粘膜输送提供了极好的机会。然而，这条路线 免疫接种在提供疫苗方面一直未得到充分利用，部分原因是唾液中存在可 会降低疫苗的质量。对于基于蛋白质的疫苗来说，情况尤其如此。在这里，我们的目标是口述 使用无针装置(叙利亚喷射器)作为口服疫苗递送系统的黏膜免疫诱导 口腔和肠道粘膜部位有较强的免疫反应。这种喷射器可以将疫苗送入口腔 因此，粘膜将加强疫苗对局部淋巴组织的输送，从而促进DC的摄取 以及2)将通过防止口服蛋白酶降解来保持疫苗的完整性。测试我们的 假设，我们将使用由CD40L佐剂DNA、MVA和 表达新城疫病毒免疫原的蛋白质疫苗。在我们的临床前猕猴研究中，我们发现CD40L- 通过IM途径交付的佐剂DNA/MVA SIV疫苗增强了对直肠内感染的保护 抗中和SIV挑战。添加纳米颗粒佐剂的系统蛋白Boost 包裹TLR7/8的DNA/MVA疫苗激动剂对血清中HIV包膜特异性免疫球蛋白的增强作用 也增加了阴道和直肠分泌物中的抗体。在本提案中，在目标1中，我们将首先优化 口服疫苗的投放方式和位置，比较不同佐剂对蛋白免疫原的影响。我们会 比较舌下免疫和口腔免疫在使用和不使用Syrijet的情况下诱导强SHV-V的作用。 口腔和肠道粘膜组织中的特异性体液免疫和细胞免疫。我们还将检测其他粘膜 蛋白质免疫佐剂鞭毛素和dmLT。在目标2中，使用最佳的给药方法和佐剂 结合我们在目标1中确定的，我们将使用分支C Shiv进行直肠内挑战研究。这 研究还将侧重于了解免疫相关因素以进行保护。通过完成这些目标，我们 希望找到一种有效的黏膜疫苗接种方案，以加强对艾滋病毒的保护。

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.