A high-throughput microparticle microarray platform for dendritic cell-targeting vaccines
A high-throughput microparticle microarray platform for dendritic cell-targeting vaccines
复制标题
DOI：
10.1016/j.biomaterials.2009.04.032
复制
发表时间：
2009-09-01
期刊：
影响因子：
14
通讯作者：
Keselowsky, Benjamin G.
中科院分区：
文献类型：
--
作者：
Acharya, Abhinav P.;Clare-Salzler, Michael J.;Keselowsky, Benjamin G.
免疫基因组学方法结合佐剂免疫学的进展正在指导疫苗的合理设计。此外，药物递送平台（例如，合成颗粒）显示出增加疫苗效力的前景。目前，存在许多已知的抗原表位和佐剂，以及许多可用于配制用于各种应用的疫苗的合成递送系统。然而，缺乏有效的手段来测试免疫细胞对大量可用组合的反应，这代表了新疫苗开发的重大障碍。为了克服这一障碍，我们报告制造的一类新的微阵列组成的抗原/抗肿瘤剂-聚（D，L丙交酯-共-乙交酯）微粒（PLGA MP），确定为一个有前途的载体免疫治疗，这是共定位与树突状细胞（DC），免疫系统的关键调节和疫苗的主要目标。其目的是利用这种高通量平台来优化基于颗粒的疫苗，该疫苗旨在体内靶向DC以治疗免疫系统相关疾病，如自身免疫性疾病，癌症和感染。DC/MP阵列的制造利用标准接触印刷微型阵列设备的使用结合表面改性以实现粒子/细胞在孤立岛上的共定位，同时提供背景非粘性表面以防止岛外细胞迁移。我们优化了MP overspotting针直径，占对齐误差，允许建设大型，高保真阵列。证明了每个点少至16+/-2 MP的可再现定量递送，并构建了双组分MP给药阵列，实现了与制剂无关的MP递送，交叉污染最小。此外，定量的斑点，表面吸附MP降解被证明，可能用于优化MP释放性能。最后，我们证明了DC与PLGA MP在孤立岛屿上的共定位，并且DC在岛屿之间不迁移长达24小时。使用这个平台，我们打算分析调制的DC功能，通过提供多参数的组合线索的形式，蛋白质，肽和其他免疫调节分子封装或拴在MP。重要的是，所获得的小型化能够通过将对细胞和试剂的需求降低许多倍来实现稀有细胞群体的高通量研究，从而促进体内靶向DC的个性化疫苗的进展。(C)2009爱思唯尔有限公司保留所有权利。
Immunogenomic approaches combined with advances in adjuvant immunology are guiding progress toward rational design of vaccines. Furthermore, drug delivery platforms (e.g., synthetic particles) are demonstrating promise for increasing vaccine efficacy. Currently there are scores of known antigenic epitopes and adjuvants, and numerous synthetic delivery systems accessible for formulation of vaccines for various applications. However, the lack of an efficient means to test immune cell responses to the abundant combinations available represents a significant blockade on the development of new vaccines. In order to overcome this barrier, we report fabrication of a new class of microarray consisting of antigen/adjuvant-loadable poly(D,L lactide-co-glycolide) microparticles (PLGA MPs), identified as a promising carrier for immunotherapeutics, which are co-localized with dendritic cells (DCs), key regulators of the immune system and prime targets for vaccines. The intention is to utilize this high-throughput platform to optimize particle-based vaccines designed to target DCs in vivo for immune system-related disorders, such as autoimmune diseases, cancer and infection. Fabrication of DC/MP arrays leverages the use of standard contact printing miniarraying equipment in conjunction with surface modification to achieve co-localization of particles/cells on isolated islands while providing background non-adhesive surfaces to prevent off-island cell migration. We optimized MP overspotting pin diameter, accounting for alignment error, to allow construction of large, high-fidelity arrays. Reproducible, quantitative delivery of as few as 16+/-2 MPs per spot was demonstrated and two-component MP dosing arrays were constructed, achieving MP delivery which was independent of formulation, with minimal cross-contamination. Furthermore, quantification of spotted, surface-adsorbed MP degradation was demonstrated, potentially useful for optimizing MP release properties. Finally, we demonstrate DC co-localization with PLGA MPs on isolated islands and that DCs do not migrate between islands for up to 24 h. Using this platform, we intend to analyze modulation of DC function by providing multi-parameter combinatorial cues in the form of proteins, peptides and other immuno-modulatory molecules encapsulated in or tethered on MPs. Critically, the miniaturization attained enables high-throughput investigation of rare cell populations by reducing the requirement for cells and reagents by many-fold, facilitating advances in personalized vaccines which target DCs in vivo. (C) 2009 Elsevier Ltd. All rights reserved.