Department of Health Sciences, Wilfrid Laurier University, Waterloo, ON, Canada.
Glysantis Inc., Guelph, ON, Canada.
Sci Rep. 2019 Sep 20;9(1):13619. doi: 10.1038/s41598-019-49931-2.
Innate immunity is induced when pathogen-associated molecular patterns (PAMPs) bind host pattern recognition receptors (PRRs). Polyinosinic:polycytidylic acid [poly(I:C)] is a synthetic analogue of viral dsRNA that acts as a PAMP, inducing type I interferons (IFNs) in vertebrates. In the present study, the immunostimulatory effects of high molecular weight (HMW) poly(I:C) in rainbow trout cells were measured when bound to a cationic phytoglycogen nanoparticle (Nano-HMW). The physical characteristics of the nanoparticle itself, when bound to different lengths of dsRNA and when cell associated was evaluated. Optimal concentration and timing for innate immune stimulation was measured using the RTG-P1 reporter cell line. The immunostimulatory effects of HMW poly (I:C) was compared to Nano-HMW in vitro using the RTgutGC cell line cultured in a conventional monolayer or a transwell culture system. The ability of an activated intestinal epithelium to transmit an antiviral signal to macrophages was evaluated using a co-culture of RTgutGC cells and RTSll (a monocyte/macrophage cell). In all culture conditions, Nano-HMW was a more effective inducer of IFN-related antiviral immune responses compared to HMW poly (I:C) alone. This study introduces the use of cationic phytoglycogen nanoparticles as a novel delivery system for immunomodulatory molecules to enhance immune responses in aquatic vertebrates.
当病原体相关分子模式 (PAMPs) 与宿主模式识别受体 (PRRs) 结合时,会诱导先天免疫。聚肌苷酸:聚胞苷酸[poly(I:C)]是一种病毒 dsRNA 的合成类似物,作为一种 PAMP,在脊椎动物中诱导 I 型干扰素 (IFNs)。在本研究中,当与阳离子植物糖纳米颗粒 (Nano-HMW) 结合时,测量了高分子量 (HMW) poly(I:C) 在虹鳟鱼细胞中的免疫刺激作用。评估了纳米颗粒本身的物理特性,以及与不同长度的 dsRNA 结合时和与细胞结合时的特性。使用 RTG-P1 报告细胞系测量了先天免疫刺激的最佳浓度和时间。使用传统单层或 Transwell 培养系统培养的 RTgutGC 细胞系,在体外比较了 HMW poly (I:C) 和 Nano-HMW 的免疫刺激作用。使用 RTgutGC 细胞和 RTSll(单核细胞/巨噬细胞细胞)的共培养物评估了激活的肠上皮向巨噬细胞传递抗病毒信号的能力。在所有培养条件下,与单独的 HMW poly (I:C) 相比,Nano-HMW 是诱导 IFN 相关抗病毒免疫反应的更有效诱导剂。本研究介绍了使用阳离子植物糖纳米颗粒作为免疫调节分子的新型递送系统,以增强水生脊椎动物的免疫反应。