Project description:We used RNAseq to investigate innate immune responsiveness in canine 030-D cell line induced by inactivated L. interrogans serogroups Canicola and Icterohaemorrhagiae, and two bivalent, non-adjuvanted canine Leptospira vaccines containing the same serogroups. We identified more than 900 DEGs associated with pathways related to innate immune responses in common to these three stimuli. Several molecules including CXCL-10, SAA, and complement factor C3 were identified that could serve as targets for development of a biomarker-based in vitro assay to assess Leptospira vaccine quality. In vitro assay could replace the current animal vaccine-challenge potency assay and contribute to reduction of animal use in vaccine manufacturing.
Project description:A mixed-aerosol pH1N1 (Cal04) challenge of rhesus macaques was establised to serve as a pre-clinical model for the evaluation of candidate vaccines. After characterizing the clinical signs and immune responses associated with pH1N1 challenge in naïve rhesus macaques, a follow-up study assessing 2 candidate vaccines was performed. This study has 2 phases: 1) Model Establisment consisting of 3 groups: Unvaccinated Live Challenge (n=3, Unvaccinated UV-inactivated Challenge (n=3), Previously Vaccinated Live Challenge (n=3) which were sampled at 2 baseline timepoints Day -7 and Day 0. Following the H1N1 Challenge, samples were collected at day 1,2,5,8,14,20. 2) Candidate Vaccine Assessment consisting of four groups: Previously Vaccinted with anti-CD40-NP5+PolyICLC (n=4), Previously vaccinated with CD40-HA+PolyICLC (n=4), Previously vaccinated with commercial mismatched Fluzone (n=4), Previously vaccinated with Media+PolyICLC alone (n=4). Daseline samples were collected at Day -7 and 0 (baseline) and Day 1,3,6,14,20 post-challenge.
Project description: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 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 canine 030-D cells stimulated with Leptospira strains, and the corresponding vaccine revealed more than 900 DEGs and 23 DEPs in common to these three stimuli. Second, comparison of responses induced by this Leptospira vaccine and a vaccine from another manufacturer revealed a large overlap in DEGs and DEPs as well, suggesting potential to identify biomarkers of Leptospira vaccine activity. 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 in the future. Additionally, this study contributes to the understanding of the mechanisms by which Leptospira vaccines induce innate immune responses in the dog.