High-Throughput Evaluation of Dendritic Cell-Targeting Vaccine Particles for the

描述（由申请人提供）：这项工作代表了一个基于生物材料的生物医学工程研究计划，与免疫学相结合，旨在实现耐受性。具体而言，该项目的重点是工程技术，以提供个性化的高通量筛选免疫细胞对基于微粒的疫苗的反应，使用有限数量的细胞。基于微粒的疫苗系统可以在体内将抗原和相关免疫调节因子递送至靶向吞噬细胞群体，具体地，树突状细胞，一种关键的免疫调节剂。对诱导耐受性疫苗的典型评估依赖于一次测试一种制剂，希望发现能够产生长期免疫耐受性的单一因素。然而，多个关键信号可能联合收割机以促进稳健的、持久的抗原特异性耐受。缺乏对不同免疫调节因子之间相互作用的理解，以及缺乏测试大量因子组合的有效手段，是新疫苗技术开发的重大障碍。为了克服这一障碍，我们正在开发一种高通量的基于细胞的微阵列方法，用于检测包含靶向树突状细胞（一种关键的抗原呈递细胞类型）的多种组分的微粒。我们的初步数据表明，我们正在开发的独特的高通量体外平台是可行的，并且微粒制剂的体外筛选可用于提示对注射微粒的体内反应。我们的长期试验床应用是通过注射微粒在糖尿病小鼠模型中预防1型糖尿病。我们正在优化多组分颗粒制剂以将DC导向致耐受性表型并诱导调节性T细胞用于抗原特异性免疫抑制。我们的小型化技术只需要少量的细胞，朝着个性化疫苗的发展迈出了一步。 公共卫生相关性：我们正在体外快速评估和优化抗原递送、免疫调节微粒作为基于微粒的可注射疫苗，旨在通过树突状细胞在体内靶向摄取，用于未来治疗1型糖尿病的研究。我们的体外系统包括制造基于细胞的免疫细胞微阵列，用于微粒制剂的高通量筛选，并且将评估制剂产生免疫细胞表型的能力，所述免疫细胞表型与诱导抗原特异性耐受有关。这种小型化的方法只使用少量的细胞，并朝着个性化疫苗的开发方向发展，这种疫苗可以针对患者的特异性免疫细胞反应进行筛选。

DESCRIPTION (provided by applicant): This work represents a biomaterials-based biomedical engineering research program integrated with immunology directed toward tolerance. Specifically, this project focuses on the engineering of technologies to provide personalized high-throughput screening of immune cell response to microparticle-based vaccines, using a limited number of cells. Microparticle-based vaccine systems can, in vivo, deliver antigen and relevant immuno- modulatory factors to targeted phagocytic cell population, specifically, dendritic cells, a key immune regulator. Typical assessment of a tolerance-inducing vaccine relies on testing one formulation at a time, hoping to uncover a single factor capable of generating long-lived immune tolerance. However, multiple critical signals are likely to combine to promote robust, enduring antigen-specific tolerance. A lack of understanding of the interactions between different immunomodulatory factors, and the lack of an efficient means to test large numbers of combinations of factors represents a significant blockade for the development of new vaccine technologies. In order to overcome this barrier, we are developing a high-throughput cell-based microarray approach for the testing of microparticles incorporating multiple components targeted to dendritic cells, a key antigen presenting cell type. Our preliminary data indicates that the unique high- throughput in vitro platform we are developing is feasible, and that in vitro screening of microparticle formulations can be useful for suggesting in vivo responses to injected microparticles. Our long-term test-bed application is the prevention of type-1 diabetes in a diabetic mouse model by injection of microparticles. We are optimizing multi- component particle formulations to direct DCs toward a tolerogenic phenotype and the induction of regulatory T-cells for antigen-specific immune suppression. Our miniaturized technology requires only small numbers of cells, taking steps toward the development of personalized vaccines. PUBLIC HEALTH RELEVANCE: We are rapidly in vitro assessing and optimizing antigen-delivering, immuno-modulatory microparticles as an injectable microparticle-based vaccine, intended for targeted uptake in vivo by dendritic cells for future studies for the treatment of type 1 diabetes. Our in vitro system consists of fabricating cell-based microarrays of immune cells for high-throughput screening of microparticle formulations, and formulations will be assessed for their ability to generate immune cell phenotypes which have been linked to the induction of antigen-specific tolerance. This miniaturized approach uses only a small number of cells, and moves toward the development of personalized vaccines, which may be screened for a patient's specific immune cell response.