Mechanism of protective immunity against Vibrio anguillarum infection in zebrafish bath-vaccinated with MVAV6203
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ABSTRACT: MVAV6203, an live attenuated vaccine showed a strong protection against V. anguillarum for Paralichthys olivaceus, Epinephelus coioides and zebrafish. However the mechanism of its protection, especially the mucosal-related response induced by immersion vaccination, is not fully understood. This study was conducted to reveal the changes of genes involved in both innative and adaptive immunity. For gene expression analysis, spleen of zebrafish from 3 vaccinated groups and 3 control groups were hybridized on a 4M-CM-^W44K aglient whole zebrafsih genome oligo microarray. Functional analysis of the microarray data was performed using KEGG and Gene Ontology (GO) analysis. Microarray gene expression analysis showed that the Th17-like immune response may play vital role in orchestrating the mucosal barrier against pathogen. Zebrafish were randomly divided into three vaccinated groups and three control groups. V.anguillarum MVAV6203 was cultured in high-salt Luria (LB) medium at 30 for 16h. The cells were harvested by centrifugation and rinsed twice in 2% saline. The desired number of cells was adjusted to 1.0E+08 CFU/mL with 2% saline. Six groups of 70 zebrafish were immersed in the aerated cell-resuspended saline or 2% saline for 10min at 24M-bM-^DM-^C. At 28 days post-vaccination pool of spleen tissue of 10 zebrafish from each group was harvested for microarray hybridization.
Project description:MVAV6203, an live attenuated vaccine showed a strong protection against V. anguillarum for Paralichthys olivaceus, Epinephelus coioides and zebrafish. However the mechanism of its protection, especially the mucosal-related response induced by immersion vaccination, is not fully understood. This study was conducted to reveal the changes of genes involved in both innative and adaptive immunity. For gene expression analysis, spleen of zebrafish from 3 vaccinated groups and 3 control groups were hybridized on a 4×44K aglient whole zebrafsih genome oligo microarray. Functional analysis of the microarray data was performed using KEGG and Gene Ontology (GO) analysis. Microarray gene expression analysis showed that the Th17-like immune response may play vital role in orchestrating the mucosal barrier against pathogen.
Project description:Mice were vaccinated with different Lentivirus vector-plaforms or PBS. Transcriptomes of sorted dendritic cells 6 hours after vaccination were used to build a model predictive of the vector-platform efficacy measured as the antigen-specific response 5, 7 and 10 days after the vaccination Groups of 7-week-old female C57BL/6 mice (Charles River, France and Germany) were immunized either intravenously or sub-cutaneously with a controlled quantity of vector particles as defined in CompuVac assay protocols (www.compuvac.eu). Control mice were injected with 100 µL of phosphate buffered saline solution (PBS).
Project description:In this study we attempt to elucidate some of the pathways involved in the immune response to vaccination and subsequent disease challenge using a transcriptomic approach. We exposed, via intra-peritoneal injection, three months old Asian seabass (Lates calcarifer) to a Streptococcus iniae vaccine. A control group was also set up where no vaccine was injected. Spleen and head kidney samples were collected at one and seven days post vaccination for transcriptomic analysis. At this point, there are four groups per organ: Day 1 vaccinated, Day 1 control, Day 7 vaccinated and Day 7 control. Subsequently, a pathogen challenge was carried out three weeks later and spleen and head kidneys were sampled at 25-29 hours post challenge for transcriptomic analysis. For control, mock challenged was carried out. At this point, there are four groups per organ: unvaccinated challenged, unvaccinated mock challenged, vaccinated challenged and vaccinated mock challenged. Total 57 samples. Spleen samples: At Day 1 and Day 7 post vaccination, 4 spleens were analyzed for each of the D1 control and D1 vaccinated groups and 3 spleens were analyzed for each of the D7 control and D7 vaccinated groups (total = 14 samples). At post pathogen challenge, 4 spleens were analyzed for each of these three groups - unvaccinated mock challenged, vaccinated challenged and vaccinated mock challenged and 3 spleens for the unvaccinated challenged group (total = 15 samples). Head Kidney samples: At Day 1 and Day 7 post vaccination, 3 head kidneys were analyzed for the control and vaccinated groups for both time points (total = 12 samples). At post pathogen challenge, 4 head kidneys were analyzed for each of the groups: unvaccinated challenged, unvaccinated mock challenged, vaccinated challenged and vaccinated mock challenged (total = 16 samples).
Project description:Systemic vaccination with the attenuated virus SIVmac239-∆Nef provides sterilizing or partial protection to rhesus monkeys challenged with WT SIV strains, providing important opportunities to study key immunological components of a protective host response. Here we show that intravenous vaccination with SIVmac239-∆Nef provides two potentially crucial immunological barriers localized at mucosal surfaces that correlate with the vaccine’s protective effects against WT SIVmac251 vaginal challenge: 1) a conditioned and coordinated response from the mucosal epithelium that blunts the early inflammatory and chemotactic signalling cascade that aids virus propagation and expansion; 2) early on-site generation/diversification of SIV-specific Abs from ectopic germinal center-like lymphoid aggregates. This unique host response to WT SIVmac251 in the female reproductive tract of SIVmac239-∆Nef-vaccinated animals points to a multi-layered strategy for a protective host response during immunodeficiency virus exposure—rapid induction of humroal immunity at mucosal surfaces without the deleterious inflammatory side effects tied to innate recognition of virus. This vaccine-induced host response highlights potential key protective mechanisms needed for an effective HIV vaccine Total RNA was isolated from the cervix of 17 Indian Rhesus macaques (3 uninfected animals; 5 unvaccinated animals 4-5 days post vaginal exposure with SIVmac251; 4 SIVmac239-∆Nef-vaccinated animals before challenge; 5 SIVmac239-∆Nef-vaccinated animals 4-5 days post vaginal exposure with SIVmac251) and prepared for hybridization on Affymetrix GeneChip Rhesus Macaque Genome Arrays. Replicate arrays were performed for a number of the samples to minimize assay noise and significant host genes altered during virus exposure in female reproductive tract tissue were identified by their associated q-values (< 0.2) and fold change in expression (> 1.2).
Project description:Salmon alphavirus (SAV) and Moritella viscosa causing respectively pancreatic disease and winter ulcer are among the most important pathogens threatening Atlantic salmon aquaculture. Fish is protected by vaccination with different rate of success. Here, responses to vaccination were assessed followed with pathogen challenges of vaccinated salmon and saline injected control.
Project description:Sequential window acquisition of all theoretical mass spectra (SWATH-MS) requires a spectral library to extract quantitative measurements from the mass spectrometry data acquired in data-independent acquisition mode (DIA). Large combined spectral libraries containing SWATH assays have been generated for humans and several other organisms, but so far no publicly available library exists for measuring the proteome of zebrafish, a rapidly emerging model system in biomedical research. Here, we present a large zebrafish SWATH spectral library to measure the abundance of 104’185 proteotypic peptides from 10’405 proteins. The library includes proteins expressed in 9 different zebrafish tissues (brain, eye, heart, intestine, liver, muscle, ovaries, spleen, and testes) and provides an important new resource to quantify 40% of the protein-coding zebrafish genes.
Project description:Pre-vaccination SARS-CoV-2 infection can boost protection elicited by COVID-19 vaccination and post-vaccination breakthrough SARS-CoV-2 infection can boost existing immunity conferred by COVID-19 vaccination. Such ‘hybrid immunity’ is effective against SARS-CoV-2 variants. In order to understand ‘hybrid immunity’ at the molecular level we studied the complementarity determining regions (CDR) of anti-RBD (receptor binding domain) antibodies isolated from individuals with ‘hybrid immunity’ as well as from ‘naive’ (notSARS-CoV-2 infected) vaccinated individuals. CDR analysis was done by liquid chromatography/mass spectrometry-mass spectrometry.
Project description:Crimean-Congo hemorrhagic fever (CCHF), caused by Crimean-Congo hemorrhagic fever virus (CCHFV), is on the World Health Organizations list over emerging pathogens and prioritized diseases. With global distribution, high fatality rate and no approved treatment or vaccine, CCHF constitute a treat against the global health. In the current study we show full protection of mice against lethal CCHFV infection due to mRNA-LNP vaccination. IFNAR-/- mice received two immunizations with either mRNA-LNP encoding for the CCHFV nucleoprotein, glycoproteins or a combination of both. While unvaccinated mice showed clear signs of severe disease, vaccinated mice was significantly protected. Vaccine induced immune responses due to vaccination was evaluated both in IFNAR-/- and immunocompetent mice and a strong humoral and cellular immune response was observed in both mouse models with high titers of neutralizing antibodies and primed T-cells. In addition, we conducted a proteomic analysis on liver samples from vaccinated and unvaccinated mice after CCHFV infection to determine the effect of vaccination on the protein profile. Similar to what has been observed in humans due to vaccination, there was an effect on metabolic pathways. In conclusion, this study shows very promising results regarding development of a vaccine against CCHFV.
Project description:Systemic vaccination with the attenuated virus SIVmac239-∆Nef provides sterilizing or partial protection to rhesus monkeys challenged with WT SIV strains, providing important opportunities to study key immunological components of a protective host response. Here we show that intravenous vaccination with SIVmac239-∆Nef provides two potentially crucial immunological barriers localized at mucosal surfaces that correlate with the vaccine’s protective effects against WT SIVmac251 vaginal challenge: 1) a conditioned and coordinated response from the mucosal epithelium that blunts the early inflammatory and chemotactic signalling cascade that aids virus propagation and expansion; 2) early on-site generation/diversification of SIV-specific Abs from ectopic germinal center-like lymphoid aggregates. This unique host response to WT SIVmac251 in the female reproductive tract of SIVmac239-∆Nef-vaccinated animals points to a multi-layered strategy for a protective host response during immunodeficiency virus exposure—rapid induction of humroal immunity at mucosal surfaces without the deleterious inflammatory side effects tied to innate recognition of virus. This vaccine-induced host response highlights potential key protective mechanisms needed for an effective HIV vaccine
Project description:Memory T and B cells in lymphoid and mucosal tissues maintain long-term protection, though their generation following vaccination remains challenging to assess in humans. Here, we investigated immune memory generated to COVID-19 mRNA vaccines across blood, lymphoid organs, and lungs from 42 vaccinated organ donors aged 23-86, of whom 57% were previously infected with SARS-CoV-2. Using high-dimensional profiling, we reveal that Spike (S)-reactive memory T cells distribute in lymphoid organs and lungs, variably express tissue resident markers based on infection history, and exhibit site-specific compositions of effector and regulatory memory T cells. S-reactive B cells are mostly class-switched memory cells localized to lymphoid organs correlating with circulating antibodies. Importantly, tissue memory T cells are more stably maintained post-vaccination and over age and exhibit a bias towards regulatory cell functional profiles compared to circulating populations. Our results show that mRNA vaccines can induce heterogeneous memory populations across sites for protection and controlling immune pathology.