Acharya Abhinav P, Carstens Matthew R, Lewis Jamal S, Dolgova Natalia, Xia C Q, Clare-Salzler Michael J, Keselowsky Benjamin G
J. Crayton Pruitt Family Department of Biomedical Engineering, University of Florida, 130 BME/PO Box 116131, Gainesville, Florida, 32611-6131, USA; Department of Materials Science and Engineering, University of Florida, USA.
J. Crayton Pruitt Family Department of Biomedical Engineering, University of Florida, 130 BME/PO Box 116131, Gainesville, Florida, 32611-6131, USA.
J Mater Chem B. 2016 Mar 7;4(9):1672-1685. doi: 10.1039/C5TB01754H. Epub 2015 Sep 30.
Experimental vaccine adjuvants are being designed to target specific toll-like receptors (TLRs) alone or in combination, expressed by antigen presenting cells, notably dendritic cells (DCs). There is a need for high-content screening (HCS) platforms to explore how DC activation is affected by adjuvant combinations. Presented is a cell-based microarray approach, "immunoarray", exposing DCs to a large number of adjuvant combinations. Microparticles encapsulating TLR ligands are printed onto arrays in a range of doses for each ligand, in all possible dose combinations. Dendritic cells are then co-localized with physisorbed microparticles on the immunoarray, adherent to isolated islands surrounded by a non-fouling background, and DC activation is quantified. Delivery of individual TLR ligands was capable of eliciting high levels of specific DC activation markers. For example, either TLR9 ligand, CpG, or TLR3 ligand, poly I:C, was capable of inducing among the highest 10% expression levels of CD86. In contrast, MHC-II expression in response to TLR4 agonist MPLA was among the highest, whereas either MPLA or poly I:C, was capable of producing among the highest levels of CCR7 expression, as well as inflammatory cytokine IL-12. However, in order to produce robust responses across all activation markers, adjuvant combinations were required, and combinations were more represented among the high responders. The immunoarray also enables investigation of interactions between adjuvants, and each TLR ligand suggested antagonism to other ligands, for various markers. Altogether, this work demonstrates feasibility of the immunoarray platform to screen microparticle-encapsulated adjuvant combinations for the development of improved and personalized vaccines.
实验性疫苗佐剂旨在单独或联合靶向抗原呈递细胞（尤其是树突状细胞，DCs）表达的特定Toll样受体（TLRs）。需要高内涵筛选（HCS）平台来探索佐剂组合如何影响DC激活。本文介绍了一种基于细胞的微阵列方法“免疫阵列”，将DCs暴露于大量佐剂组合中。将包裹TLR配体的微粒以每种配体的一系列剂量打印到阵列上，形成所有可能的剂量组合。然后将树突状细胞与免疫阵列上物理吸附的微粒共定位，这些微粒附着在由防污背景包围的孤立岛屿上，并对DC激活进行定量。递送单个TLR配体能够引发高水平的特异性DC激活标志物。例如，TLR9配体CpG或TLR3配体聚肌苷酸胞苷酸（poly I:C）能够诱导CD86表达水平处于最高的10%之中。相比之下，响应TLR4激动剂单磷酰脂质A（MPLA）的MHC-II表达处于最高水平，而MPLA或poly I:C能够产生最高水平的CCR7表达以及炎性细胞因子IL-12。然而，为了在所有激活标志物上产生强烈反应，需要佐剂组合，并且高反应者中组合的情况更为常见。免疫阵列还能够研究佐剂之间的相互作用，并且每种TLR配体对于各种标志物都显示出与其他配体的拮抗作用。总之，这项工作证明了免疫阵列平台筛选微粒包裹的佐剂组合以开发改进型和个性化疫苗的可行性。
