Project description:Female mice aged 6-7 weeks old were infected with the T. cruzi H1 strain. Mice were randomly divided into groups with 15 animals in each, with age matched naïve mice included as controls. Approximately 75 days post infection, mice were randomly divided into groups and treated with different treatment combinations: BNZ + Tc24 therapeutic vaccine + the STAT-3 inhibitor TTI-101, BNZ + Tc24 vaccine, and BNZ or TTI alone. Mice were humanely euthanized at 50, 75, 120 or 142 days post infection (DPI), and hearts samples were collected, divided into left atrium (LA), right atrium (RA), left ventricle top (LVT), left ventricle bottom (LVB), right ventricle top (RVT), right ventricle bottom (RVB). Heart tissue samples were extracted by 1:1 (v/v) methanol to water for aqueous phase and 3:1 (v/v) dichloromethane to methanol for organic phase. LC-MS/MS data acquisition was performed on a Q Exactive Plus Hybrid Quadrupole-Orbitrap Mass Spectrometer coupled to a Thermo Scientific Vanquish UHPLC with 1.7 um 100 Å Kinetex C8 column at 40°C. Mobile phase A was water with 0.1% (v/v) formic acid and mobile phase B was acetonitrile with 0.1% (v/v) formic acid.
Project description:Female mice aged 6-7 weeks old were infected with the T. cruzi H1 strain. Mice were randomly divided into groups with 15 animals in each, with age matched naïve mice included as controls. Approximately 75 days post infection, mice were randomly divided into groups and treated with different treatment combinations: BNZ + Tc24 therapeutic vaccine + the STAT-3 inhibitor TTI-101, BNZ + Tc24 vaccine, and BNZ or TTI alone. Mice were humanely euthanized at 50, 75, 120 or 142 days post infection (DPI), and hearts samples were collected, divided into left atrium (LA), right atrium (RA), left ventricle top (LVT), left ventricle bottom (LVB), right ventricle top (RVT), right ventricle bottom (RVB). Heart tissue samples were extracted by 1:1 (v/v) methanol to water for aqueous phase and 3:1 (v/v) dichloromethane to methanol for organic phase. LC-MS/MS data acquisition was performed on a Q Exactive Plus Hybrid Quadrupole-Orbitrap Mass Spectrometer coupled to a Thermo Scientific Vanquish UHPLC with 1.7 um 100 Å Kinetex C8 column at 40°C. Mobile phase A was water with 0.1% (v/v) formic acid and mobile phase B was acetonitrile with 0.1% (v/v) formic acid.
Project description:Vaccine development involves time-consuming and expensive evaluation of candidate vaccines in animal models. As mediators of both innate and adaptive immune responses dendritic cells (DCs) are considered to be highly important for vaccine performance. Here we evaluated in how far the response of DCs to a vaccine in vitro is in line with the immune response the vaccine evokes in vivo. To this end, we investigated the response of murine bone marrow-derived DCs to whole inactivated virus (WIV) and subunit (SU) influenza vaccine preparations. These vaccine preparations were chosen because they differ in the immune response they evoke in mice with WIV being superior to SU vaccine through induction of higher virus-neutralizing antibody titers and a more favorable Th1-skewed response phenotype. To evaluate if in vivo immunogenicity is reflected by DC reactions in vitro we studied the gene expression signature of murine bone-marrow-derived conventional DCs (cDCs) upon stimulation with WIV or SU influenza vaccine or, for reasons of comparison, with live influenza virus. Dendritic cells stimulated with PBS served as a control. Gene expression analysis was performed on samples 4, 12 and 24 hours after the start of stimulation.
Project description:Results from the Step trial in humans and studies in non-human primates highlighted a role for heightened activated CD4 T cell response in promoting HIV/SIV acquisition. However, the contribution of vaccine-specific CD4 T cell response in influencing protection is not known. Here, using the macaque model, we show that vaccine-induced Th1-biased CCR5+ CD4 T cell response in blood and mucosal tissue above a certain thresh¬old is detrimental for vaccine-mediated protection against pathogenic mucosal SIV infections.
Project description:Mice were immunized with either formalin fixed Influenza A/PR/8/34 (Killed PR8), the 2006-2007 seasonal influenza vaccine, the 2007-2008 seasonal influenza vaccine, a sublethal infection (live PR8) or mock immunized (PBS). Array data was used to distinguish the immunogens from each other and predict which of the three inactivated vaccines would be protective against A/PR/8/34 challenge. two replicates of each peptide was printed on 1 CIM_10kv3 peptide microarray. One microarray were tested for each sample. Image was qualified using in-house metrics for quality assurance.
Project description:To compare the effects of different types of SARS-CoV-2 vaccines in booster immunization, this study established a population cohort vaccinated with inactivated vaccine and protein subunit vaccine as the third booster vaccine, respectively. We collected serum and PBMC samples from participants in chronological order, and performed a systematic review of distinct antibody signatures and microtranscriptomics to provide data support for booster immunization.
Project description:Live-attenuated viral vaccines have been successfully used to combat infectious disease for decades but due to their empirical derivation, little is known about their mechanisms of attenuation. This lack of understanding makes the development of next generation live attenuated vaccines difficult. The success of the 17D vaccine and availability of the parent virus, Asibi, makes it an excellent model to understand the molecular basis of attenuation of a live attenuated vaccine and the effects of viral diversity on vaccine function. Due to the differences in genetic diversity between WT Asibi virus and its 17D vaccine derivative, we investigated the changes in genetic diversity of 17D and Asibi viruses following treatment with ribavirin.
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.