Biomarkers for canine Leptospira vaccine potency
Ontology highlight
ABSTRACT:
PROVIDER: PRJNA793869 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| SRR17413122.fastq.gz | Fastqsanger.gz | |||
| SRR17413123.fastq.gz | Fastqsanger.gz | |||
| SRR17413124.fastq.gz | Fastqsanger.gz | |||
| SRR17413125.fastq.gz | Fastqsanger.gz | |||
| SRR17413126.fastq.gz | Fastqsanger.gz |
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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.
2022-03-06 | PXD031875 | Pride
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.
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Project description:Identification of Biomarkers of Vaccine Reactogenicity
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Project description:The overall goal of these experiments was to determine how human endothelial cells respond to pathogenic Leptospira interrogans. Leptospira interrogans causes leptospirosis, the most widespread zoonotic infection in the world. A hallmark of leptospirosis is widespread endothelial damage, which in severe cases leads to hemorrhage. In these experiments, we infected two endothelial cell lines with pathogenic Leptospira interrogans serovar Canicola strain Ca12-005, and as controls, with the non-pathogenic Leptospira biflexa serovar Patoc strain Pfra. As additional controls, uninfected cells were also included in the analyses.
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Project description:The overall goal of these experiments was to determine how human endothelial cells respond to pathogenic Leptospira interrogans. Leptospira interrogans causes leptospirosis, the most widespread zoonotic infection in the world. A hallmark of leptospirosis is widespread endothelial damage, which in severe cases leads to hemorrhage. In these experiments, we infected two endothelial cell lines with pathogenic Leptospira interrogans serovar Canicola strain Ca12-005, and as controls, with the non-pathogenic Leptospira biflexa serovar Patoc strain Pfra. As additional controls, uninfected cells were also included in the analyses.
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Project description:BioVacSafe (Biomarkers for enhanced vaccine immunosafety) Influenza challenge
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Project description:Systems vaccinology approaches have been used to successfully define early signatures of the vaccine-induced immune response. However, the possibility that transcriptomics can also predict vaccine reactogenicity has not been fully explored. We have compared four licensed vaccines that have a demonstrated safety profile, as well as three agonists of TLRs that have a known inflammatory potential, to elucidate the transcriptomic profile of an acceptable response to a vaccination versus those which push the inflammatory envelope. In mice, we looked at the transcriptomic changes in the injected muscle, the lymph node that drained the muscle and the PBMC isolated from the circulating blood from the very early timepoint of 4 hours to over the period of one week. A detailed examination and comparative analysis of the transcriptome from each of these tissues, at all the timepoints of 4, 8, 24, 48, 72 and 168 hr and with these eight different vaccines, TLR or saline control injections, revealed a set of novel biomarkers that are reflective of inflammation after vaccination. These biomarkers are easily sampled and readily measurable in the peripheral blood, providing a useful tool to predict levels of reactogenicity, as a way to select candidates with acceptable immunogenicity and safety profiles.
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Project description:Pathogenic Leptospira spp. are the causative agents of the zoonotic disease leptospirosis. During infection, Leptospira are confronted with deadly reactive oxygen species (ROS). Withstanding ROS produced by the host innate immunity is an important strategy evolved by pathogenic Leptospira for persisting in and colonizing hosts. The peroxide stress regulator, PerR, represses genes involved in ROS defenses in L. interrogans. We have performed RNA sequencing in WT and perR mutant strains to characterize the L. interrogans adaptive response to hydrogen peroxide. We showed that Leptospira solicit three main peroxidase machineries (catalase, cytochrome C peroxidase and peroxiredoxin) and heme to adapt to peroxide stress as well as canonical chaperones of the heat shock response, and DNA repair. Determining the PerR regulon allowed to identify the PerR-dependent mechanisms of the peroxide adaptive response and has revealed a regulatory network involving other transcriptional regulators, two-component systems and sigma factors as well as non-coding RNAs that putatively orchestrate, in concert with PerR, this adaptive response. Our findings provide comprehensive insight into the mechanisms required by pathogenic Leptospira to overcome infection-related oxidants. This will participate in framing future hypothesis-driven studies to identify and decipher novel virulence mechanisms.
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Project description:Leptospirosis, caused by bacteria of the genus Leptospira, is a zoonotic disease affecting humans, companion animals, and all major livestock species. Typical propagation of the highly fastidious Leptospira borgepetesenii serovar Hardjo is limited to 29°C. However, newer culture media formulations now facilitate isolation and propagation at 37°C, a temperature that more closely emulates in vivo conditions and is hypothesized to regulate the expression of virulence factors during host infection. Since protein expression by leptospires is temperature dependent, and therefore the proteome of bacterin vaccines can differ whether grown at 37°C compared to 29°C, we compared the proteome of strains of Leptospira borgpetersenii serovar Hardjo at each temperature; two well-established strains that causes acute (strain JB197) or chronic asymptomatic disease (strain HB203) in the hamster challenge model of leptospirosis and two more recently isolated strains designated TC129 and TC273 (both of which cause chronic asymptomatic disease in the hamster). We found proteomic expression differences within strains propagated at the routine temperature of 29°C, and compared to the newly achieved culture temperature of 37°C. Results highlight significant differential protein expression, including virulence factors, amongst identical serovars of L. borgpetersenii when propagated at 29oC, the collective variation of which can be diminished when propagated at 37oC. Collectively, there is increasingly more evidence available to suggest bacterin vaccine design would benefit from consideration of strains employed, and potential effects of growth temperature related to specific behavior of pathogens in vaccine composition.
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Project description:Urothelial carcinoma (UC) is the most common tumour type in canine bladder cancer. Current diagnostic methods are technically challenging or can lack specificity, hence there is a need for novel biomarkers of UC. To this end, we analysed the microRNA (miRNA) cargo of extracellular vesicles (EVs) from urine samples of dogs with UC to identify miRNAs that could be utilised as biomarkers. Urine was fractionated using ultrafiltration combined with size-exclusion chromatography and small RNA sequencing analysis was performed on both the EV enriched and (EV free) protein fractions. A greater number of candidate miRNA biomarkers were detected in the EV fraction than the protein fraction, and further validation using droplet digital PCR (ddPCR) was performed on the EV enriched fraction of a second cohort of dogs with bladder cancer which validated three miRNAs as candidate biomarkers; miR-182, miR-221 and miR-222 as being significantly overrepresented in dogs with UC when compared with healthy dogs and dogs with urinary tract infections. Pathway analysis confirmed that these three miRNAs are involved in cancer. In addition, their potential downstream gene targets were predicted and PIK3R1, a well-known oncogene is likely to be a shared target between miRNA-182 and miRNA-221/222. In summary, this study highlights the potential of urinary EV-associated miRNAs as a source of biomarkers for the diagnosis of canine UC.
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