Department of Clinical Analysis, School of Pharmaceutical Sciences, São Paulo State University (UNESP), Araraquara 14800-903, São Paulo, Brazil.
Pharmacology Department, Faculty of Medicine, Universitat de Valencia, 46010 Valencia, Spain.
Int J Mol Sci. 2021 Mar 27;22(7):3470. doi: 10.3390/ijms22073470.
In recent years, there has been great interest in developing molecular adjuvants based on antisense oligonucleotides (ASOs) targeting immunosuppressor pathways with inhibitory effects on regulatory T cells (Tregs) to improve immunogenicity and vaccine efficacy. We aim to evaluate the immunostimulating effect of 2'OMe phosphorothioated Foxp3-targeted ASO in an antifungal adjuvanted recombinant vaccine.
The uptake kinetics of Foxp3 ASO, its cytotoxicity and its ability to deplete Tregs were evaluated in murine splenocytes in vitro. Groups of mice were vaccinated with recombinant enolase (Eno) of in Montanide Gel 01 adjuvant alone or in combination with either 1 µg or 8 µg of Foxp3 ASO. The titers of antigen-specific antibody in serum samples from vaccinated mice (male C57BL/6) were determined by ELISA (enzyme-linked immunosorbent assay). Cultured splenocytes from each group were activated in vitro with Eno and the levels of IFN-γ and IL-12 were also measured by ELISA. The results showed that the anti-Eno antibody titer was significantly higher upon addition of 8 µM Foxp3 ASO in the vaccine formulation compared to the standard vaccine without ASO. In vitro and in vivo experiments suggest that Foxp3 ASO enhances specific immune responses by means of Treg depletion during vaccination.
Foxp3 ASO significantly enhances immune responses against co-delivered adjuvanted recombinant Eno vaccine and it has the potential to improve vaccine immunogenicity.
近年来，人们对开发基于免疫抑制途径的反义寡核苷酸（ASO）作为分子佐剂产生了浓厚的兴趣，这些 ASO 对调节性 T 细胞（Tregs）具有抑制作用，旨在提高免疫原性和疫苗效力。我们旨在评估靶向 Foxp3 的 2'OMe 硫代磷酸化 ASO 在抗真菌佐剂重组疫苗中的免疫刺激作用。
在体外评估 Foxp3 ASO 的摄取动力学、细胞毒性及其耗尽 Tregs 的能力。将重组烯醇化酶（Eno）单独或与 1µg 或 8µg Foxp3 ASO 联合在 Montanide Gel 01 佐剂中免疫雄性 C57BL/6 小鼠。通过酶联免疫吸附试验（ELISA）测定血清样品中抗原特异性抗体的滴度。用 Eno 体外激活每组培养的脾细胞，并通过 ELISA 测量 IFN-γ 和 IL-12 的水平。结果表明，与不含 ASO 的标准疫苗相比，疫苗制剂中添加 8µM Foxp3 ASO 可显著提高抗 Eno 抗体滴度。体外和体内实验表明，Foxp3 ASO 通过在接种过程中耗尽 Treg 来增强特异性免疫反应。
Foxp3 ASO 可显著增强针对共递呈佐剂重组 Eno 疫苗的免疫反应，具有提高疫苗免疫原性的潜力。
