In the last few years, interest has grown in the use of nucleic acids as an ocular therapy for retinal genetic diseases. Recently, our research group has demonstrated that mRNA delivery could result in effective protein expression in ocular cells following subretinal injection. Yet, although mRNA therapy comes with many advantages, its immunogenicity resulting in hampered mRNA translation delays development to the clinic. Therefore, several research groups investigate possible strategies to reduce this innate immunity. In this study, we focus on B18R, an immune inhibitor to suppress the mRNA-induced innate immune responses in two ocular cell types. We made use of retinal pigment epithelial (RPE) cells and Müller cells both as immortalized cell lines and primary bovine cells. When cells were co-incubated with both B18R and mRNA-MessengerMAX lipoplexes we observed an increase in transfection efficiency accompanied by a decrease in interferon-β production, except for the Müller cells. Moreover, uptake efficiency and cell viability were not hampered. Taken together, we showed that the effect of B18R is cell type-dependent but remains a possible strategy to improve mRNA translation in RPE cells.

中文翻译：

---

痘苗病毒蛋白 B18R：不同眼细胞类型中非病毒传递对 mRNA 免疫原性和翻译的影响

在过去的几年中，人们对使用核酸作为视网膜遗传疾病的眼部疗法的兴趣日益浓厚。最近，我们的研究小组证明，视网膜下注射后，mRNA 递送可以在眼细胞中产生有效的蛋白质表达。然而，尽管mRNA疗法具有许多优点，但其免疫原性导致mRNA翻译受阻，延迟了临床开发。因此，几个研究小组研究了减少这种先天免疫的可能策略。在这项研究中，我们重点关注 B18R，这是一种免疫抑制剂，可抑制两种眼细胞类型中 mRNA 诱导的先天免疫反应。我们使用视网膜色素上皮 (RPE) 细胞和 Müller 细胞作为永生化细胞系和原代牛细胞。当细胞与 B18R 和 mRNA-MessengerMAX lipoplex 共孵育时，我们观察到转染效率增加，同时干扰素-β 产量减少（Müller 细胞除外）。此外，摄取效率和细胞活力没有受到阻碍。综上所述，我们发现 B18R 的作用具有细胞类型依赖性，但仍然是改善 RPE 细胞中 mRNA 翻译的可能策略。