Novel recombinant Rotavirus vaccine utilizing the probiotic microorganism Lactobacillus acidophilus

项目摘要/摘要 利用益生菌嗜酸乳杆菌构建新型重组轮状病毒疫苗 尽管2006年两种轮状病毒疫苗在全球范围内获得许可并可用，但据估计，轮状病毒 每年杀死大约215,000名儿童。发展中国家的5岁以下儿童 更有可能受到严重轮状病毒病的影响，因为目前可用的疫苗 与发达国家(85%)相比，有效率(50%-60%)。持续的发病率和死亡率以及 当前减毒活疫苗带来的风险，需要开发下一代人类 轮状病毒疫苗。我们已经开发了一种口服粘膜疫苗平台，它使用乳杆菌 嗜酸菌。我们将创新的佐剂和抗原表达策略结合在一起 被证明具有诱导强大的粘膜和系统免疫反应的潜力的构建体。这个平台 提供了几个重要的可行性优势，因为它采用了指定为GRAS(通常 被FDA认为是安全的)，生产成本低，不需要冷链，并且是无针的。我们会 利用适应嗜酸乳杆菌的新型CRISPR-Cas9系统来设计复杂的疫苗结构， 将使用同型小鼠轮状病毒挑战进行测试。同型和异型中和的诱导 抗体将使用一种新的、高度敏感的分析方法进行评估。我们将评估疫苗株的定植情况 以及对微生物组的潜在偏离目标的影响。最后，我们将直接比较我们的RLA 轮状病毒疫苗对抗人活疫苗的动物实验研究 移植人婴儿粪便微生物组。针对人类轮状病毒挑战的有效性将被确定。 在完成这些拟议的研究后，我们的目标是准备好RLA构建，以便向人类进行 临床试验。

PROJECT SUMMARY / ABSTRACT Novel recombinant Rotavirus vaccine utilizing the probiotic microorganism Lactobacillus acidophilus Despite the worldwide licensure and availability of two rotavirus vaccines in 2006, it is estimated that rotavirus kills approximately 215,000 children each year. Children under the age of 5 in developing countries are significantly more likely to be affected by severe rotavirus disease as the currently available vaccines have lower efficacy (50-60%) as compared to developed nations (>85%). The ongoing morbidity and mortality as well as the risk posed by current live-attenuated vaccines, necessitate the development of a next generation human rotavirus vaccine. We have developed an orally-delivered, mucosal vaccine platform that employs Lactobacillus acidophilus. We have brought together innovative adjuvant and antigen-expression strategies in unique constructs with demonstrated potential to induce robust mucosal and systemic immune responses. This platform offers several important feasibility advantages as it employs a commensal designated as GRAS (generally regarded as safe) by the FDA, is inexpensive to produce, does not require cold-chain, and is needleless. We will utilize a novel CRISPR-Cas9 system adapted to L. acidophilus to engineer sophisticated vaccine constructs that will be tested using a homotypic mouse rotavirus challenge. Induction of homotypic and heterotypic neutralizing antibodies will be assessed using a novel, highly sensitive assay. We will assess vaccine strain colonization and potential off-target effects on the microbiome. Finally, we will perform a direct comparison of our rLA rotavirus vaccine against a human live attenuated vaccine in the gnotobiotic pig model using animals transplanted with human infant fecal microbiome. Efficacy against human rotavirus challenge will be determined. At the completion of these proposed studies, we aim to have a rLA construct ready to proceed toward human clinical trials.