Division of Infectious Diseases and Immunology, Department of Biomolecular Health Sciences, Faculty of Veterinary Medicine, Utrecht University, Utrecht, The Netherlands.
Intravacc, Bilthoven, The Netherlands.
Sci Rep. 2022 Aug 4;12(1):13418. doi: 10.1038/s41598-022-16457-z.
Mandatory potency testing of Leptospira vaccine batches relies partially on in vivo procedures, requiring large numbers of laboratory animals. Cell-based assays could replace in vivo tests for vaccine quality control if biomarkers indicative of Leptospira vaccine potency are identified. We investigated innate immune responsiveness induced by inactivated L. interrogans serogroups Canicola and Icterohaemorrhagiae, and two bivalent, non-adjuvanted canine Leptospira vaccines containing the same serogroups. First, the transcriptome and proteome analysis of a canine monocyte/macrophage 030-D cell line stimulated with Leptospira strains, and vaccine B revealed more than 900 DEGs and 23 DEPs in common to these three stimuli. Second, comparison of responses induced by vaccine B and vaccine D revealed a large overlap in DEGs and DEPs as well, suggesting potential to identify biomarkers indicative of Leptospira vaccine quality. Because not many common DEPs were identified, we selected seven molecules from the identified DEGs, associated with pathways related to innate immunity, of which CXCL-10, IL-1β, SAA, and complement C3 showed increased secretion upon stimulation with both Leptospira vaccines. These molecules could be interesting targets for development of biomarker-based assays for Leptospira vaccine quality control in the future. Additionally, this study contributes to the understanding of the mechanisms by which Leptospira vaccines induce innate immune responses in the dog.
疫苗批次的强制性效力测试部分依赖于体内程序,需要大量的实验动物。如果能确定指示莱姆病疫苗效力的生物标志物,则可以用基于细胞的测定法来替代体内试验,用于疫苗质量控制。我们研究了犬单核细胞/巨噬细胞 030-D 细胞系对灭活的钩端螺旋体血清群堪萨斯和伊氏出血群,以及含有相同血清群的两种非佐剂的二价犬用莱姆病疫苗产生的固有免疫反应。首先,用莱姆螺旋体株和疫苗 B 刺激犬单核细胞/巨噬细胞 030-D 细胞系,对其进行转录组和蛋白质组分析,发现这三种刺激物有超过 900 个 DEGs 和 23 个 DEPs 是共同的。其次,比较疫苗 B 和疫苗 D 诱导的反应,发现 DEGs 和 DEPs 也有很大的重叠,这表明有可能确定指示莱姆病疫苗质量的生物标志物。由于很少有共同的 DEPs 被识别,我们从鉴定的 DEGs 中选择了七个与先天免疫途径相关的分子,其中 CXCL-10、IL-1β、SAA 和补体 C3 在受到两种莱姆病疫苗刺激时,分泌量增加。这些分子可能是未来开发基于生物标志物的莱姆病疫苗质量控制测定法的有趣靶标。此外,本研究有助于了解莱姆病疫苗在犬中诱导固有免疫反应的机制。