Immunotherapeutic nanoparticles: Implications for the treatment of tuberculosis and HIV

据估计，人类携带者患结核病的风险要高出26到31倍 免疫缺陷病毒(HIV-1)。结核病的艰苦治疗方案(鸡尾酒至少6个月) 具有剂量限制副作用的抗生素)几十年来基本上没有变化。减少了 全球结核病负担主要集中在新的抗生素和疫苗上，对靶向巨噬细胞的关注很少 激活免疫系统以增加微生物清除的药物输送方法。来解决这个未得到满足的问题 根据需要，我们开发了一种靶向巨噬细胞疗法，可广泛应用于艾滋病毒和结核病。我们的配方 将模型利福平药物β-葡聚糖和壳聚糖包被的聚乳酸-羟基乙酸 (Glu-CS-PLGA)纳米粒子。我们的药代动力学研究发现，在单个纳米颗粒 通过口咽吸入给药，在健康小鼠的细胞片段中检测到利福平。 给药后最多10天进行支气管肺泡灌洗。此外，我们还发现了肿瘤坏死因子α的释放时间过程， 在给药后24小时恢复到基线。所有纳米粒在一定范围内都能刺激肿瘤坏死因子α的释放。 在500至2000pg/ml之间，与药物治疗后结核病小鼠模型中肿瘤坏死因子α的浓度相似。当我们有了 使用我们的纳米颗粒证明了利福平在小鼠肺泡巨噬细胞中的持续递送，我们已经 然而，要了解这种纳米颗粒如何在强毒结核分枝杆菌的环境中发挥作用 阿龙。此外，我们还没有发现这种纳米制剂在缺乏的情况下如何影响结核肉芽肿。 或HIV病毒蛋白的存在。为了解决这个问题，我们正在采取双管齐下的方法，独立 目标。目的1，在体外结核肉芽肿模型中测定GLU-CS-PLGA纳米粒 的存在，将分析GLU-CS-PLGA纳米粒子在 模特。我们还包括了一项探索性实验，以研究HIV-1蛋白对 Glu-CS-PLGA纳米粒的药效。目的2，测定GLU-CS-PLGA纳米粒在小鼠体内的药效 BALB/c结核小鼠模型“，将在下列情况下在肺内原位测定杀菌效果 GLU-CS-PLGA口咽抽吸治疗BALB/c小鼠结核模型因此，这个纳米平台是 与影响巨噬细胞的疾病广泛相关，包括结核病和艾滋病毒(艾滋病毒巨噬细胞储存库)。 确定这种纳米配方的有效性质对于决定是否(1)我们应该放弃 这种治疗方法或者(2)我们应该继续在先进的小鼠结核病模型中测试它 高度组织化的包裹性坏死性病变，即肉芽肿，含有大量细胞外 这种细菌更类似于人类的病变，在艾滋病毒-结核病混合感染模型中。这是一项创新 提出了促进免疫系统治疗的概念性和机械性知识 传染病和这种新型免疫刺激药物输送纳米颗粒平台。通过将 当谷氨酸进入纳米颗粒的表面时，产生的免疫刺激不依赖于其治疗货物。

The risk of developing tuberculosis (TB) is estimated to be 26 to 31 times greater in people living with Human Immunodeficiency Virus (HIV-1). The arduous treatment regimen for TB (minimum of six months on a cocktail of antibiotics with dose-limiting side-effects) has remained essentially unchanged for decades. Reducing the global TB burden has focused primarily on new antibiotics and vaccines with little focus on macrophage targeted drug delivery methods that activate the immune system to increase microbial eradication. To address this unmet need, we developed a targeted macrophage therapy with broad applications for HIV and TB. Our formulation delivers a model rifamycin drug, rifampin, loaded into β-glucan and chitosan coated poly(lactic-co-glycolic) acid (GLU-CS-PLGA) nanoparticles. Our pharmacokinetic studies found that after a single nanoparticle administration via oropharyngeal aspiration to healthy mice, rifampin was detected in the cellular fraction of the bronchoalveolar lavage up to 10 days post-dosing. Furthermore, we found a time course of release of TNFα that returns to baseline at 24 hr post administration. All nanoparticles stimulated the release of TNFα in the range of 500 to 2000 pg/ml, similar to TNFα concentrations in a TB mouse model after drug treatment. While we have demonstrated sustained delivery of rifampin to alveolar macrophage in mice using our nanoparticles, we have yet to learn how this nanoparticle functions in the setting of virulent mycobacterium tuberculosis in the lung. Furthermore, we have yet to discover how this nanoformulation affects a TB granuloma in the absence or presence of HIV viral proteins. To address this, we are taking a two-pronged approach with independent aims. Aim 1, “Determine the GLU-CS-PLGA nanoparticles in an in vitro TB granuloma model in the absence or presence of HIV proteins”, will analyze the bactericidal efficacy elicited by GLU-CS-PLGA nanoparticles in this model. We also have included an exploratory experiment to investigate the effects of HIV-1 proteins on GLU-CS-PLGA nanoparticle efficacy. Aim 2, “Determine the in vivo efficacy of GLU-CS-PLGA nanoparticles in the BALB/c TB mouse model”, will determine the bactericidal efficacy in situ in the lungs following oropharyngeal aspiration with GLU-CS-PLGA in the BALB/c TB mouse model. Therefore, this nanoplatform is broadly relevant to disorders that affect macrophage, including TB and HIV (HIV macrophage reservoirs). Determining the efficacious nature of this nanoformulation is essential to decide whether (1) we should abandon this therapeutic approach or (2) we should move forward to test it in an advanced mouse TB model that develops highly organized encapsulated necrotic lesions, i.e., granulomas, that contain large numbers of extracellular bacilli that more closely resembled human lesions and, in an HIV-TB co-infection model. This innovative proposal advances the conceptual and mechanistic knowledge of stimulation of the immune system to treat infectious diseases and this novel immune-stimulating drug delivery nanoparticle platform. By incorporating GLU onto the nanoparticle's surface, the resulting immune stimulation is independent of its therapeutic cargo.