Lewis Jamal S, Roche Chris, Zhang Ying, Brusko Todd M, Wasserfall Clive H, Atkinson Mark, Clare-Salzler Michael J, Keselowsky Benjamin G
J. Crayton Pruitt Family Department of Biomedical Engineering, University of Florida, Gainesville, FL 32611.
Department of Pathology, Immunology and Laboratory Medicine, University of Florida, Gainesville, FL 32611.
J Mater Chem B. 2014 May 7;2(17):2562-2574. doi: 10.1039/C3TB21460E.
Microparticulate systems are beginning to show promise for delivery of modulatory agents for immunotherapeutic applications which modulate dendritic cell (DC) functions. Co-administration of multiple factors is an emerging theme in immune modulation which may prove beneficial in this setting. Herein, we demonstrate that localized, controlled delivery of multiple factors can be accomplished through poly (lactic-co-glycolic acid) (PLGA) microparticle systems fabricated in two size classes of phagocytosable and unphagocytosable microparticles (MPs). The immunosuppressive ability of combinatorial multi-factor dual MP systems was evaluated by investigating effects on DC maturation, DC resistance to LPS-mediated maturation and proliferation of allogeneic T cells in a mixed lymphocyte reaction. Phagocytosable MPs (2 μm) were fabricated encapsulating either rapamycin (RAPA) or all-trans retinoic acid (RA), and unphagocytosable MPs (30 μm) were fabricated encapsulating either transforming growth factor beta-1 (TGF-β1) or interleukin-10 (IL-10). Combinations of these MP classes reduced expression of stimulatory/costimulatory molecules (MHC-II, CD80 and CD86) in comparison to iDC and soluble controls, but not necessarily to single factor MPs. Dual MP-treated DCs resisted LPS-mediated activation, in a manner driven by the single factor phagocytosable MPs used. Dendritic cells treated with dual MP systems suppressed allogeneic T cell proliferation, generally demonstrating greater suppression by combination MPs than single factor formulations, particularly for the RA/IL-10 MPs. This work demonstrates feasibility of simultaneous targeted delivery of immunomodulatory factors to cell surface receptors and intracellular locations, and indicates that a combinatorial approach can boost immunoregulatory responses for therapeutic application in autoimmunity and transplantation.
微粒系统在递送用于免疫治疗应用的调节因子方面开始展现出前景，这些调节因子可调节树突状细胞（DC）的功能。多种因子的联合给药是免疫调节中一个新兴的主题，在这种情况下可能被证明是有益的。在此，我们证明通过聚（乳酸-乙醇酸）（PLGA）微粒系统可以实现多种因子的局部、可控递送，该系统由可吞噬和不可吞噬的两种尺寸类别的微粒（MPs）制成。通过研究对DC成熟、DC对LPS介导的成熟的抗性以及混合淋巴细胞反应中同种异体T细胞增殖的影响，评估了组合多因子双MP系统的免疫抑制能力。制备了可吞噬的MPs（约2μm），包封雷帕霉素（RAPA）或全反式维甲酸（RA），以及不可吞噬的MPs（约30μm），包封转化生长因子β-1（TGF-β1）或白细胞介素-10（IL-10）。与未成熟DC和可溶性对照相比，这些MP类别组合降低了刺激/共刺激分子（MHC-II、CD80和CD86）的表达，但不一定比单因子MPs降低得更多。双MP处理的DC以所用单因子可吞噬MPs驱动的方式抵抗LPS介导的激活。用双MP系统处理的树突状细胞抑制同种异体T细胞增殖，通常组合MPs比单因子制剂表现出更大的抑制作用，特别是对于RA/IL-10 MPs。这项工作证明了将免疫调节因子同时靶向递送至细胞表面受体和细胞内位置的可行性，并表明组合方法可以增强免疫调节反应，用于自身免疫和移植的治疗应用。
