Bernal Laura, Alvarado-Vázquez Abigail, Ferreira David Wilson, Paige Candler A, Ulecia-Morón Cristina, Hill Bailey, Caesar Marina, Romero-Sandoval E Alfonso
Department of Pharmaceutical and Administrative Sciences, Presbyterian College School of Pharmacy, 307 N. Broad St, Clinton, SC 29325, USA; Department of Systems' Biology, School of Medicine, University of Alcala. Campus Universitario - C/ 19, Carretera Madrid-Barcelona, Km 33,600, 28871 Alcalá de Henares, Madrid, Spain.
Department of Pharmaceutical and Administrative Sciences, Presbyterian College School of Pharmacy, 307 N. Broad St, Clinton, SC 29325, USA.
Immunobiology. 2017 Feb;222(2):399-408. doi: 10.1016/j.imbio.2016.08.010. Epub 2016 Sep 5.
Macrophages orchestrate the initiation and resolution of inflammation by producing pro- and anti-inflammatory products. An imbalance in these mediators may originate from a deficient or excessive immune response. Therefore, macrophages are valid therapeutic targets to restore homeostasis under inflammatory conditions. We hypothesize that a specific mannosylated nanoparticle effectively induces gene expression in human macrophages under inflammatory conditions without undesirable immunogenic responses. THP-1 macrophages were challenged with lipopolysaccharide (LPS, 5μg/mL). Polyethylenimine (PEI) nanoparticles grafted with a mannose receptor ligand (Man-PEI) were used as a gene delivery method. Nanoparticle toxicity, Man-PEI cellular uptake rate and gene induction efficiency (GFP, CD14 or CD68) were studied. Potential immunogenic responses were evaluated by measuring the production of tumor necrosis factor-alpha (TNF-α), Interleukin (IL)-6 and IL-10. Man-PEI did not produce cytotoxicity, and it was effectively up-taken by THP-1 macrophages (69%). This approach produced a significant expression of GFP (mRNA and protein), CD14 and CD68 (mRNA), and transiently and mildly reduced IL-6 and IL-10 levels in LPS-challenged macrophages. Our results indicate that Man-PEI is suitable for inducing an efficient gene overexpression in human macrophages under inflammatory conditions with limited immunogenic responses. Our promising results set the foundation to test this technology to induce functional anti-inflammatory genes.
巨噬细胞通过产生促炎和抗炎产物来协调炎症的起始和消退。这些介质的失衡可能源于免疫反应不足或过度。因此，巨噬细胞是在炎症条件下恢复体内平衡的有效治疗靶点。我们假设一种特定的甘露糖基化纳米颗粒在炎症条件下能有效诱导人巨噬细胞中的基因表达，且不会产生不良免疫原性反应。用脂多糖（LPS，5μg/mL）刺激THP-1巨噬细胞。将接枝有甘露糖受体配体的聚乙烯亚胺（PEI）纳米颗粒（Man-PEI）用作基因递送方法。研究了纳米颗粒毒性、Man-PEI细胞摄取率和基因诱导效率（绿色荧光蛋白、CD14或CD68）。通过测量肿瘤坏死因子-α（TNF-α）、白细胞介素（IL）-6和IL-10的产生来评估潜在的免疫原性反应。Man-PEI未产生细胞毒性，并且能被THP-1巨噬细胞有效摄取（69%）。这种方法使绿色荧光蛋白（mRNA和蛋白）、CD14和CD68（mRNA）显著表达，并使LPS刺激的巨噬细胞中IL-6和IL-10水平短暂且轻度降低。我们的结果表明，Man-PEI适合在炎症条件下诱导人巨噬细胞中高效的基因过表达，且免疫原性反应有限。我们的 promising 结果为测试该技术以诱导功能性抗炎基因奠定了基础。
