A STINGing vaccine for TB

项目摘要 结核病（TB）是全球单一传染病病原体导致死亡的主要原因。疫苗接种 是任何传染病最具成本效益的控制干预措施。Bacille Calmette-Guérin（BCG） 这是人类历史上使用最广泛的疫苗，但就目前使用而言，它未能控制结核病。因此 因此，开发改进的结核病疫苗仍然是全球高度优先事项。最近的研究表明 卡介苗，当修改，通过替代途径管理，或用于复种，提供改善， 保护，这表明它已经准备好卷土重来。 我们已经产生了一种新的重组BCG称为BCG-disA-OE， c-di-AMP，一种有效的STING激动剂我们的初步研究表明，BCG-disA-OE比 BCG-WT（野生型BCG）在预防TB攻击后的疾病（POD）以及作为免疫疗法 非肌肉浸润性膀胱癌（NMIBC），其中膀胱内BCG是目前的一线治疗。 接种BCG-disA-OE的豚鼠对气雾剂攻击的保护作用明显更好， 我们发现BCG-disA-OE在大鼠中也显示出上级功效 和NMIBC的小鼠模型。与BCG-WT相比，BCG-disA-OE导致更有效的促炎作用， 巨噬细胞和膀胱癌细胞的细胞因子反应，更高程度的促炎表观遗传 标记，以及更大的髓样细胞向M1表型极化-这些变化都与 增强的“训练免疫”，一种新发现的现象，其特征在于表观遗传和功能 先天免疫细胞的重编程。 在本申请中，我们的中心科学前提是，将STING激动剂过表达添加到 BCG将增强巨噬细胞中训练有素的免疫变化，并提供更有效的抗结核保护。 为了验证这些假设，在目标1中，我们将确定BCG-disA-OE与BCG-WT的保护效果。 针对非人灵长类动物（NHP）中的TB疾病和针对小鼠中潜伏性结核病的再活化。在 目的2：评价两株卡介苗诱导巨噬细胞系训练免疫功能的变化， 以及原代鼠、NHP和人（来自健康供体的白细胞）巨噬细胞。我们将包括串行 从血液和NHP的BAL以及获得的死后组织中采集NHP单核细胞 在目标1下。在目标3中，我们将描述结核病中骨髓和淋巴细胞群体的极化特征。 在TB的小鼠和NHP模型中，预先接种BCG-WT与BCG-disA-OE诱导的肉芽肿。

PROJECT SUMMARY Tuberculosis (TB) is the leading cause of death from a single infectious disease agent worldwide. Vaccination is the most cost-effective control intervention for any infectious disease. Bacille Calmette–Guérin (BCG) remains the most widely used vaccine in human history, but as currently used it has failed to control TB. Thus, the development of improved vaccines against TB therefore remains a high global priority. Recent studies indicate that BCG, when modified, administered through alternate routes, or used in revaccination, offers improved protection, suggesting that it is well poised to make comeback. We have generated a novel recombinant BCG known as BCG-disA-OE which is engineered to overexpress c-di-AMP, a potent STING agonist Our preliminary studies show that BCG-disA-OE is more effective than BCG-WT (wild type BCG) in prevention of disease (POD) following TB challenge and also as an immunotherapy for non-muscle invasive bladder cancer (NMIBC) where intravesical BCG is currently the first-line therapy. Guinea pigs vaccinated with BCG-disA-OE were significantly better protected against aerosol challenge with virulent M.tb than with BCG-WT, and we found that BCG-disA-OE also showed superior efficacy BCG-WT in rat and mouse models of NMIBC. Compared with BCG-WT, BCG-disA-OE leads to more potent pro-inflammatory cytokine responses in macrophage and bladder cancer cells, a higher degree of proinflammatory epigenetic marks, and greater myeloid cell polarization towards the M1 phenotype—changes that are all consistent with enhanced “trained immunity”, a newly discovered phenomenon characterized by epigenetic and functional reprogramming of innate immune cells. In this application, our central scientific premise is that the addition of STING agonist overexpression to BCG will augment trained immunity changes in macrophages and provide more effective protection against TB. To test these hypotheses, in Aim 1 we will determine the protective efficacy of BCG-disA-OE versus BCG-WT against TB disease in non-human primates (NHPs) and against reactivation of latent tuberculosis in mice. In Aim 2 we will evaluate trained immunity changes induced by the two BCG strains in macrophage cell lines, as well as primary murine, NHP, and human (leukopacs from healthy donors) macrophages. We will include serial sampling of NHP mononuclear cells from blood and BAL of NHPs as well as their post-mortem tissues obtained under Aim 1. In Aim 3, we will characterize polarization of myeloid and lymphoid cell populations in the TB granuloma induced by prior vaccination with BCG-WT versus BCG-disA-OE in mouse and NHP models of TB.