Boehler R M, Kuo R, Shin S, Goodman A G, Pilecki M A, Gower R M, Leonard J N, Shea L D
Department of Chemical and Biological Engineering, Northwestern University, Evanston, IL.
Biotechnol Bioeng. 2014 Jun;111(6):1210-21. doi: 10.1002/bit.25175. Epub 2014 Feb 12.
Gene delivery from biomaterials can create an environment that promotes and guides tissue formation. However, the immune response induced upon biomaterial implantation can be detrimental to tissue regeneration. Macrophages play a central role in mediating early phases of this response, and functional "polarization" of macrophages towards M1 (inflammatory) or M2 (anti-inflammatory) phenotypes may bias the local immune state at the implant site. Since gene delivery from biomaterial scaffolds can confer transgene expression in macrophages in vivo, we investigated whether transduction of macrophages with an IL-10 encoding lentivirus can (1) induce macrophage polarization toward an M2 phenotype even in an pro-inflammatory environment, and (2) prevent a shift in polarization from M2 to M1 following exposure to pro-inflammatory stimuli. IL-10 lentivirus delivery to pre-polarized M1 macrophages reduced TNF-α production 1.5-fold when compared to cells treated with either a control virus or a bolus delivery of recombinant IL-10 protein. IL-10 lentivirus delivery to naïve macrophages reduced the amount of TNF-α produced following an inflammatory challenge by 2.5-fold compared to cells treated with both the control virus and recombinant IL-10. At a mechanistic level, IL-10 lentivirus delivery mediated sustained reduction in NF-κB activation and, accordingly, reduced transcription of TNF-α. In sum, lentiviral delivery of IL-10 to macrophages represents a promising strategy for directing and sustaining macrophage polarization towards an M2 phenotype in order to promote local immune responses that facilitate tissue engineering.
生物材料介导的基因传递能够营造出促进并引导组织形成的环境。然而，生物材料植入后引发的免疫反应可能对组织再生不利。巨噬细胞在介导该反应的早期阶段发挥着核心作用，巨噬细胞向M1（促炎）或M2（抗炎）表型的功能性“极化”可能会使植入部位的局部免疫状态产生偏差。由于生物材料支架介导的基因传递可使体内巨噬细胞实现转基因表达，我们研究了用编码白细胞介素-10（IL-10）的慢病毒转导巨噬细胞是否能够：（1）即使在促炎环境中也能诱导巨噬细胞向M2表型极化；（2）在暴露于促炎刺激后防止极化状态从M2转变为M1。与用对照病毒或重组IL-10蛋白大剂量处理的细胞相比，向预极化的M1巨噬细胞递送IL-10慢病毒可使肿瘤坏死因子-α（TNF-α）的产生减少1.5倍。与用对照病毒和重组IL-10处理的细胞相比，向未极化的巨噬细胞递送IL-10慢病毒可使炎症刺激后产生的TNF-α量减少2.5倍。在机制层面，IL-10慢病毒递送介导了核因子-κB（NF-κB）激活的持续降低，从而减少了TNF-α的转录。总之，向巨噬细胞递送IL-10慢病毒是一种很有前景的策略，可引导并维持巨噬细胞向M2表型极化，以促进有利于组织工程的局部免疫反应。
