Department of Pharmaceutical Technology, University of Regensburg, Regensburg, Germany.
Institute of Virology, Technical University of Munich / Helmholtz Munich, Munich, Germany.
Eur J Pharm Biopharm. 2023 Nov;192:41-55. doi: 10.1016/j.ejpb.2023.09.015. Epub 2023 Sep 27.
Safe and effective vaccines have been regarded early on as critical in combating the COVID-19 pandemic. Among the deployed vaccine platforms, subunit vaccines have a particularly good safety profile but may suffer from a lower immunogenicity compared to mRNA based or viral vector vaccines. In fact, this phenomenon has also been observed for SARS-CoV-2 subunit vaccines comprising the receptor-binding domain (RBD) of the spike (S) protein. Therefore, RBD-based vaccines have to rely on additional measures to enhance the immune response. It is well accepted that displaying antigens on nanoparticles can improve the quantity and quality of vaccine-mediated both humoral and cell-mediated immune responses. Based on this, we hypothesized that SARS-CoV-2 RBD as immunogen would benefit from being presented to the immune system via silica nanoparticles (SiNPs). Herein we describe the preparation, in vitro characterization, antigenicity and in vivo immunogenicity of SiNPs decorated with properly oriented RBD in mice. We found our RBD-SiNP conjugates show narrow, homogeneous particle distribution with optimal size of about 100 nm for efficient transport to and into the lymph node. The colloidal stability and binding of the antigen was stable for at least 4 months at storage- and in vivo-temperatures. The antigenicity of the RBD was maintained upon binding to the SiNP surface, and the receptor-binding motif was readily accessible due to the spatial orientation of the RBD. The particles were efficiently taken up in vitro by antigen-presenting cells. In a mouse immunization study using an mRNA vaccine and spike protein as benchmarks, we found that the SiNP formulation was able to elicit a stronger RBD-specific humoral response compared to the soluble protein. For the adjuvanted RBD-SiNP we found strong S-specific multifunctional CD4 T cell responses, a balanced T helper response, improved auto- and heterologous virus neutralization capacity, and increased serum avidity, suggesting increased affinity maturation. In summary, our results provide further evidence for the possibility of optimizing the cellular and humoral immune response through antigen presentation on SiNP.
安全有效的疫苗被视为对抗 COVID-19 大流行的关键。在已部署的疫苗平台中,亚单位疫苗具有特别良好的安全性,但与基于 mRNA 或病毒载体的疫苗相比,其免疫原性可能较低。事实上,这种现象也在包含刺突 (S) 蛋白受体结合域 (RBD) 的 SARS-CoV-2 亚单位疫苗中观察到。因此,基于 RBD 的疫苗必须依赖额外的措施来增强免疫反应。人们普遍认为,将抗原呈现在纳米颗粒上可以提高疫苗介导的体液和细胞介导免疫反应的数量和质量。基于此,我们假设 SARS-CoV-2 RBD 作为免疫原,通过硅纳米颗粒 (SiNP) 递呈给免疫系统将从中受益。在此,我们描述了在小鼠中制备、体外表征、抗原性和体内免疫原性的 SiNP 上用适当取向的 RBD 修饰的 SiNP。我们发现我们的 RBD-SiNP 缀合物具有窄且均匀的颗粒分布,最佳尺寸约为 100nm,可有效递送至淋巴结并进入淋巴结。在储存和体内温度下,抗原的胶体稳定性和结合至少稳定 4 个月。RBD 结合到 SiNP 表面后,其抗原性得以维持,并且由于 RBD 的空间取向,受体结合基序易于接近。在体外,这些颗粒被抗原呈递细胞有效摄取。在使用 mRNA 疫苗和 Spike 蛋白作为基准的小鼠免疫研究中,我们发现 SiNP 配方能够比可溶性蛋白引发更强的 RBD 特异性体液反应。对于佐剂的 RBD-SiNP,我们发现强烈的 S 特异性多功能 CD4 T 细胞反应、平衡的辅助 T 细胞反应、提高的自身和异源病毒中和能力以及增加的血清亲合力,提示亲和力成熟增加。总之,我们的结果为通过 SiNP 上的抗原呈递优化细胞和体液免疫反应的可能性提供了进一步的证据。
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