Active virosomes (AVs) are derivatives of viruses, broadly similar to ‘parent’ pathogens, with an outer envelope that contains a bespoke genome coding for four to five viral proteins capable of eliciting an antigenic response. AVs are essentially novel vaccine formulations that present on their surface selected viral proteins as antigens. Once administered, they elicit an initial ‘anti-viral’ immune response. AVs are also internalised by host cells where their cargo viral genes are used to express viral antigen(s) intracellularly. These can then be transported to the host cell surface resulting in a second wave of antigen exposure and a more potent immuno-stimulation. A new 3D correlative microscopy approach is used here to provide a robust analytical method for characterisation of Zika- and Chikungunya-derivatised AV populations including vesicle size distribution and variations in antigen loading. Manufactured batches were compared to assess the extent and nature of batch-to-batch variations. We also show preliminary results that verify antigen expression on the surface of host cells. We present here a reliable and efficient high-resolution 3D imaging regime that allows the evaluation of the microstructure and biochemistry of novel vaccine formulations such as AVs.

中文翻译：

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在低温下使用超分辨率荧光显微镜和软 X 射线断层扫描的相关成像提供了一种评估病毒体解决方案以用于疫苗开发的新方法

活性病毒体 (AVs) 是病毒的衍生物，与“亲本”病原体大体相似，具有包含定制基因组的外壳，该基因组编码四到五个能够引发抗原反应的病毒蛋白。AVs 本质上是一种新型疫苗制剂，在其表面存在选择的病毒蛋白作为抗原。一旦给药，它们就会引发最初的“抗病毒”免疫反应。AV 也被宿主细胞内化，其中它们的货物病毒基因用于在细胞内表达病毒抗原。然后这些可以被转运到宿主细胞表面，导致第二波抗原暴露和更有效的免疫刺激。这里使用一种新的 3D 相关显微镜方法来提供一种可靠的分析方法，用于表征寨卡病毒和基孔肯雅病毒衍生的 AV 种群，包括囊泡大小分布和抗原负载的变化。比较制造的批次以评估批次间差异的程度和性质。我们还展示了验证宿主细胞表面抗原表达的初步结果。我们在这里提出了一种可靠且高效的高分辨率 3D 成像方案，该方案允许评估新型疫苗制剂（如 AV）的微观结构和生物化学。