Early immune responses in skin and lymph node after skin delivery of Toll-like receptor agonists in neonatal and adult pigs

Vreman, Sandra; Rebel, Johanna M.J.; McCaffrey, Joanne; Ledl, Kristina; Arkhipova, Ksenia; Collins, Damien; McDaid, Dennis; Savelkoul, H.F.J.; Skovgaard, Kerstin; Moore, Anne; Stockhofe-Zurwieden, Norbert

doi:10.1016/j.vaccine.2021.02.028

Cited by 5 publications

(10 citation statements)

References 57 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…cDNA synthesis was performed using QuantiTect Reverse Transcription Kit (Qiagen) according to manufacturer's recommendations and as previously described (32), with an input of 500 ng total RNA. As control for genomic DNA (gDNA) contamination, samples with high RIN value were chosen as non-reverse transcription controls.…”

Section: Immune Gene Expression By High Throughput Microfluidic Qpcrmentioning

confidence: 99%

“…Pre-amplification was performed using TaqMan PreAmp Master Mix (Applied Biosystems) as previously described (32). Briefly, preamplification was performed with a 20 µM primer mix containing all assays as shown in Table 3, with the following thermal cycling conditions: 95 • C for 10 min, 20 cycles: 95 • C for 15 s and 60 • C for 4 min.…”

Section: Immune Gene Expression By High Throughput Microfluidic Qpcrmentioning

confidence: 99%

See 1 more Smart Citation

Decreased water temperature enhance Piscine orthoreovirus genotype 3 replication and severe heart pathology in experimentally infected rainbow trout

Sørensen

Cuenca²,

Olsen³

et al. 2023

Front. Vet. Sci.

Self Cite

View full text Add to dashboard Cite

Piscine orthoreovirus genotype 3 (PRV-3) was first discovered in Denmark in 2017 in relation to disease outbreaks in rainbow trout (Oncorhynchus mykiss). While the virus appears to be widespread in farmed rainbow trout, disease outbreaks associated with detection of PRV-3 have only occurred in recirculating aquaculture systems, and has predominantly been observed during the winter months. To explore the possible effects of water temperature on PRV-3 infection in rainbow trout, an in vivo cohabitation trial was conducted at 5, 12, and 18°C. For each water temperature, a control tank containing mock-injected shedder fish and a tank with PRV-3 exposed fish were included. Samples were collected from all experimental groups every 2nd week post challenge (WPC) up until trial termination at 12 WPC. PRV-3 RNA load measured in heart tissue of cohabitants peaked at 6 WPC for animals maintained at 12 and 18°C, while it reached its peak at 12 WPC in fish maintained at 5°C. In addition to the time shift, significantly more virus was detected at the peak in fish maintained at 5°C compared to 12 and 18°C. In shedders, fish at 12 and 18°C cleared the infection considerably faster than the fish at 5°C: while shedders at 18 and 12°C had cleared most of the virus at 4 and 6 WPC, respectively, high virus load persisted in the shedders at 5°C until 12 WPC. Furthermore, a significant reduction in the hematocrit levels was observed in the cohabitants at 12°C in correlation with the peak in viremia at 6 WPC; no changes in hematocrit was observed at 18°C, while a non-significant reduction (due to large individual variation) trend was observed at cohabitants held at 5°C. Importantly, isg15 expression was positively correlated with PRV-3 virus load in all PRV-3 exposed groups. Immune gene expression analysis showed a distinct gene profile in PRV-3 exposed fish maintained at 5°C compared to 12 and 18°C. The immune markers mostly differentially expressed in the group at 5°C were important antiviral genes including rigi, ifit5 and rsad2 (viperin). In conclusion, these data show that low water temperature allow for significantly higher PRV-3 replication in rainbow trout, and a tendency for more severe heart pathology development in PRV-3 injected fish. Increased viral replication was mirrored by increased expression of important antiviral genes. Despite no mortality being observed in the experimental trial, the data comply with field observations of clinical disease outbreaks during winter and cold months.

show abstract

Section: Immune Gene Expression By High Throughput Microfluidic Qpcrmentioning

confidence: 99%

Section: Immune Gene Expression By High Throughput Microfluidic Qpcrmentioning

confidence: 99%

Decreased water temperature enhance Piscine orthoreovirus genotype 3 replication and severe heart pathology in experimentally infected rainbow trout

Sørensen

Cuenca²,

Olsen³

et al. 2023

Front. Vet. Sci.

Self Cite

View full text Add to dashboard Cite

show abstract

“…Column-based extraction of RNA with DNAse treatment was performed manually with the RNeasy mini kit (Qiagen, Sweden) according to the manufacturer's protocol [56]. Duplicate cDNA synthesis and preamplification was conducted as described previously [57] using 500 ng total RNA and 19 cycles of preamplification for all four tissues. qPCR of the preamplified cDNA was performed in 192.24 Dynamic Arrays (Fluidigm), combining 192 preamplified samples with 23 primer sets (Supplementary Table S4) in 4608 individual and simultaneous qPCR reactions as described previously in Barington et al [58].…”

Section: Gene-expression Analysis Using Microfluidic Qpcrmentioning

confidence: 99%

“…qPCR data analysis was performed as described previously in Vreman et al [57] using the following validated reference genes: RPL13a and FAU (kidney, spleen, liver) and ElF1a, RPL13a, and FAU (brain). GeNorm [59] and GeNormFinder [60] were used to identify the most stable reference genes in GenEx6 (MultiD Analyses AB).…”

Section: Gene-expression Data Analysismentioning

confidence: 99%

Nervous Necrosis Virus-like Particle (VLP) Vaccine Stimulates European Sea Bass Innate and Adaptive Immune Responses and Induces Long-Term Protection against Disease

et al. 2021

Self Cite

View full text Add to dashboard Cite

The rapidly increasing Mediterranean aquaculture production of European sea bass is compromised by outbreaks of viral nervous necrosis, which can be recurrent and detrimental. In this study, we evaluated the duration of protection and immune response in sea bass given a single dose of a virus-like particle (VLP)-based vaccine. Examinations included experimental challenge with nervous necrosis virus (NNV), serological assays for NNV-specific antibody reactivity, and immune gene expression analysis. VLP-vaccinated fish showed high and superior survival in challenge both 3 and 7.5 months (1800 and 4500 dd) post-vaccination (RPS 87 and 88, OR (surviving) = 16.5 and 31.5, respectively, p < 0.01). Although not providing sterile immunity, VLP vaccination seemed to control the viral infection, as indicated by low prevalence of virus in the VLP-vaccinated survivors. High titers of neutralizing and specific antibodies were produced in VLP-vaccinated fish and persisted for at least ~9 months post-vaccination as well as after challenge. However, failure of immune sera to protect recipient fish in a passive immunization trial suggested that other immune mechanisms were important for protection. Accordingly, gene expression analysis revealed that VLP-vaccination induced a mechanistically broad immune response including upregulation of both innate and adaptive humoral and cellular components (mx, isg12, mhc I, mhc II, igm, and igt). No clinical side effects of the VLP vaccination at either tissue or performance levels were observed. The results altogether suggested the VLP-based vaccine to be suitable for clinical testing under farming conditions.

show abstract

“…Because the immune system develops during early-life, the vaccination of young animals is less successful at the moment than that of adults. In her PhD research, Sandra Vreman showed that vaccine formulations could be specified for particular ages and administration routes (Vreman et al, 2021). Skin vaccination is a promising method for triggering immune cells in pigs.…”

Section: Vaccination Strategiesmentioning

confidence: 99%

Healthy and resilient livestock

Rebel¹

2022

Preprint

View full text Add to dashboard Cite

Early immune responses in skin and lymph node after skin delivery of Toll-like receptor agonists in neonatal and adult pigs

Cited by 5 publications

References 57 publications

Decreased water temperature enhance Piscine orthoreovirus genotype 3 replication and severe heart pathology in experimentally infected rainbow trout

Decreased water temperature enhance Piscine orthoreovirus genotype 3 replication and severe heart pathology in experimentally infected rainbow trout

Nervous Necrosis Virus-like Particle (VLP) Vaccine Stimulates European Sea Bass Innate and Adaptive Immune Responses and Induces Long-Term Protection against Disease

Healthy and resilient livestock

Contact Info

Product

Resources

About