Dowlath Sasheen, Campbell Katrin, Al-Barwani Farah, Young Vivienne L, Young Sarah L, Walker Greg F, Ward Vernon K
Department of Microbiology & Immunology, School of Biomedical Sciences, University of Otago, P.O. Box 56, 720 Cumberland St, Dunedin 9054, New Zealand.
Department of Pathology, Dunedin School of Medicine, University of Otago, P.O. Box 56, 720 Cumberland St, Dunedin 9054, New Zealand.
Vaccines (Basel). 2021 Mar 3;9(3):213. doi: 10.3390/vaccines9030213.
Biologics can be combined with liquid polymer materials and electrospun to produce a dry nanofibrous scaffold. Unlike spray-drying and freeze-drying, electrospinning minimizes the physiological stress on sensitive materials, and nanofiber mat properties such as hydrophobicity, solubility, and melting temperature can be tuned based on the polymer composition. In this study, we explored the dry formulation of a virus-like particle (VLP) vaccine by electrospinning VLP derived from rabbit hemorrhagic disease virus modified to carry the MHC-I gp100 tumor-associated antigen epitope. VLP were added to a polyvinylpyrrolidone (PVP) solution (15% /) followed by electrospinning at 24 kV. Formation of a nanofibrous mat was confirmed by scanning electron microscopy, and the presence of VLP was confirmed by transmission electron microscopy and Western blot. VLP from the nanofibers induced T-cell activation and interferon- (IFN-) γ production in vitro. To confirm in vivo cytotoxicity, Pmel mice treated by injection with gp100 VLP from nanofibers induced a gp100 specific immune response, lysing approximately 65% of gp100-pulsed target cells, comparable to mice vaccinated with gp100 VLP in PBS. VLP from nanofibers also induced an antibody response. This work shows that electrospinning can be used to dry-formulate VLP, preserving both humoral and cell-mediated immunity.
生物制剂可以与液体聚合物材料结合并通过静电纺丝制备出干燥的纳米纤维支架。与喷雾干燥和冷冻干燥不同，静电纺丝能将对敏感材料的生理压力降至最低，并且纳米纤维垫的性质，如疏水性、溶解性和熔点，可以根据聚合物组成进行调整。在本研究中，我们通过静电纺丝源自经修饰携带MHC-I gp100肿瘤相关抗原表位的兔出血病病毒的病毒样颗粒（VLP），探索了VLP疫苗的干燥制剂。将VLP添加到聚乙烯吡咯烷酮（PVP）溶液（15%/）中，然后在24 kV下进行静电纺丝。通过扫描电子显微镜确认了纳米纤维垫的形成，并通过透射电子显微镜和蛋白质免疫印迹法确认了VLP的存在。纳米纤维中的VLP在体外诱导T细胞活化和干扰素-（IFN-）γ产生。为了确认体内细胞毒性，通过注射纳米纤维中的gp100 VLP处理的Pmel小鼠诱导了gp100特异性免疫反应，裂解了约65%的gp100脉冲靶细胞，这与用PBS中的gp100 VLP接种的小鼠相当。纳米纤维中的VLP还诱导了抗体反应。这项工作表明，静电纺丝可用于VLP的干燥制剂，同时保留体液免疫和细胞介导的免疫。
