A STINGing vaccine for TB

项目总结 结核病(TB)是世界范围内单一传染病病原体造成的主要死亡原因。接种疫苗 是对任何传染病最具成本效益的控制干预措施。卡介苗(BCG)残留 这是人类历史上使用最广泛的疫苗，但目前使用的它未能控制结核病。因此， 因此，开发改进的结核病疫苗仍然是全球的高度优先事项。最近的研究表明 卡介苗，当修改，通过替代途径给药，或用于复种时，提供了更好的 保护，表明它已经做好了卷土重来的准备。 我们已经产生了一种名为BCG-DISA-OE的新型重组卡介苗，它被设计成过表达 一种有效的刺痛激动剂我们的初步研究表明BCG-DISA-OE比 BCG-WT(野生型BCG)在结核病挑战后预防疾病(POD)中的作用及其作为免疫治疗的作用 对于非肌肉浸润性膀胱癌(NMIBC)，目前BCG膀胱内灌注是一线治疗方法。 接种BCG-DISA-OE的豚鼠对气雾剂攻击的保护作用显著增强 BCG-DISA-OE对大鼠的药效也优于BCG-WT。 以及NMIBC的小鼠模型。与BCG-WT相比，BCG-DISA-OE具有更强的促炎作用 巨噬细胞和膀胱癌细胞的细胞因子反应，更高程度的促炎性表观遗传 标记和更大的髓系细胞向M1表型分化-这些变化都与 增强“训练免疫”--一种表观遗传性和功能性的新发现现象 先天免疫细胞的重新编程。 在这个应用中，我们的中心科学前提是将刺痛激动剂过度表达添加到 卡介苗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病保护。 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 抗非人类灵长类(NHP)中的结核病和小鼠中潜伏的结核病的重新激活。在……里面 目的评价两株卡介苗在巨噬细胞系中诱导的训练性免疫变化。 以及原代小鼠、NHP和人类(来自健康捐赠者的白质)巨噬细胞。我们将包括序列号 NHP患者血液和BAL及其死后组织中NHP单个核细胞的采样 在目标1下。在目标3中，我们将描述结核病中髓系和淋巴系细胞群的极化 BCG-WT与BCG-DISA-OE预先接种诱发的小鼠肉芽肿和NHP结核模型。

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.