1.
Anderson, Jenna
(författare)
Development and evaluation of a subunit DIVA vaccine against bluetongue virus serotype 8 in cattle
2014
Doktorsavhandling (övrigt vetenskapligt/konstnärligt) abstract
Bluetongue virus (BTV) causes the primarily vector-borne bluetongue disease of ruminants, which poses a permanent threat to Europe since new serotypes and strains are frequently introduced. Vaccination of cattle is essential to control BTV outbreaks. Commercial attenuated and inactivated vaccines are efficacious in reducing BTV spread and disease, but do not fulfil all safety, adaptability, or production requirements. Additionally, no current vaccines allow the differentiation of infected from vaccinated animals (DIVA). DIVA vaccines enable surveillance of BTV epidemiology and vaccine efficacy, and facilitate a quick return for countries to a BTV-free status. This thesis presents the development and evaluation of a novel subunit DIVA vaccine against BTV serotype 8 (BTV-8) in cattle. Five His-tagged recombinant BTV proteins (VP2, VP5 of BTV-8; NS1, NS2, NS3 of BTV-2) were produced in baculovirus or E. coli expression systems. Purification protocols were optimized for all but VP5. Based on the feasibility of protein production and the capability of the remaining four proteins to induce humoral or cellular immune responses in mice, VP2, NS1, and NS2 were selected to formulate an experimental vaccine combined to an ISCOM-matrix adjuvant (SubV). Next, cattle were immunized twice at a three-week interval with SubV, a commercial inactivated vaccine, or a placebo. SubV induced humoral immune responses, including virus-neutralizing antibodies, against all three proteins, as well as a cellular immune response directed against NS1. These responses were of similar type and comparable magnitude between both vaccines, suggesting that SubV might provide protection that is at least as effective as the commercial vaccine. Finally, the protective efficacy of SubV was evaluated and complete virological and clinical protection against virulent BTV-8 challenge was observed following vaccination in calves. This was likely due to the induction of virus-neutralizing antibodies directed against VP2 of BTV-8 and cross-serotype T cell responses directed against NS1 and NS2 of BTV-2. Furthermore, SubV was shown to be DIVA-compliant based on the detection of antibodies directed against VP7, by using commercially-available diagnostic assays. This novel BTV subunit vaccine is a promising candidate and should be further developed.
2.
Nordmark, Gunnel, et al.
(författare)
Additive effects of the major risk alleles of IRF5 and STAT4 in primary Sjögren's syndrome
2009
Ingår i: Genes and Immunity. - : Springer Science and Business Media LLC. - 1466-4879 .- 1476-5470. ; 10:1, s. 68-76
Tidskriftsartikel (refereegranskat) abstract
Primary Sjögren's syndrome (SS) shares many features with systemic lupus erythematosus (SLE). Here we investigated the association of the three major polymorphisms in IRF5 and STAT4 found to be associated with SLE, in patients from Sweden and Norway with primary SS. These polymorphisms are a 5-bp CGGGG indel in the promoter of IRF5, the single nucleotide polymorphism (SNP) rs10488631 downstream of IRF5 and the STAT4 SNP rs7582694, which tags the major risk haplotype of STAT4. We observed strong signals for association between all three polymorphisms and primary SS, with odds ratios (ORs) >1.4 and P-values <0.01. We also found a strong additive effect of the three risk alleles of IRF5 and STAT4 with an overall significance between the number of risk alleles and primary SS of P=2.5 × 10−9. The OR for primary SS increased in an additive manner, with an average increase in OR of 1.78. For carriers of two risk alleles, the OR for primary SS is 1.43, whereas carriers of five risk alleles have an OR of 6.78. IRF5 and STAT4 are components of the type I IFN system, and our findings emphasize the importance of this system in the etiopathogenesis of primary SS.
3.
LeBlanc, Neil, et al.
(författare)
A novel combination of TaqMan RT-PCR and a suspension microarray assay for the detection and species identification of pestiviruses
2010
Ingår i: Veterinary Microbiology. - : Elsevier BV. - 0378-1135 .- 1873-2542. ; 142:1-2, s. 81-86
Tidskriftsartikel (refereegranskat) abstract
The genus pestivirus contains four recognized species: classical swine fever virus, border disease virus, bovine viral diarrhoea virus types 1 and 2. All are economically important and globally distributed but classical swine fever is the most serious, concerning losses and control measures. It affects both domestic pigs and wild boars. Outbreaks of this disease in domestic pigs call for the most serious measures of disease control, including a stamping out policy in Europe. Since all the members of the pestivirus genus can infect swine, differential diagnosis using traditional methods poses some problems. Antibody tests may lack specificity due to cross-reactions, antigen capture ELISAs may have low sensitivity, and virus isolation may take several days or even longer time to complete. PCR-based tests overcome these problems for the most part, but in general lack the multiplexing capability to detect and differentiate all the pestiviruses simultaneously. The assay platform described here addresses all of these issues by combining the advantages of real-time PCR with the multiplexing capability of microarray technology. The platform includes a TaqMan real-time PCR designed for the universal detection of pestiviruses and a microarray assay that can use the amplicons produced in the real-time PCR to identify the specific pestivirus.
4.
5.
Einarsdottir, Sigrun, et al.
(författare)
Deficiency of SARS-CoV-2 T-cell responses after vaccination in long-term allo-HSCT survivors translates into abated humoral immunity.
2022
Ingår i: Blood advances. - : American Society of Hematology. - 2473-9537 .- 2473-9529. ; 6:9, s. 2723-2730
Tidskriftsartikel (refereegranskat) abstract
Recipients of allogeneic hematopoietic stem cell transplantation (allo-HSCT) for hematological diseases are at risk of severe disease and death from COVID-19. To determine the safety and immunogenicity of BNT162b2 and mRNA-1273 COVID-19 vaccines, samples from 50 infection-naive allo-HSCT recipients (median, 92 months from transplantation, range, 7-340 months) and 39 healthy controls were analyzed for serum immunoglobulin G (IgG) against the receptor binding domain (RBD) within spike 1 (S1) of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2; anti-RBD-S1 IgG) and for SARS-CoV-2-specific T-cell immunity, reflected by induction of T-cell-derived interferon-γ in whole blood stimulated ex vivo with 15-mer SI-spanning peptides with 11 amino acid overlap S1-spanning peptides. The rate of seroconversion was not significantly lower in allo-transplanted patients than in controls with 24% (12/50) and 6% (3/50) of patients remaining seronegative after the first and second vaccination, respectively. However, 58% of transplanted patients lacked T-cell responses against S1 peptides after 1 vaccination compared with 19% of controls (odds ratio [OR] 0.17; P = .009, Fisher's exact test) with a similar trend after the second vaccination where 28% of patients were devoid of detectable specific T-cell immunity, compared with 6% of controls (OR 0.18; P = .02, Fisher's exact test). Importantly, lack of T-cell reactivity to S1 peptides after vaccination heralded substandard levels (<100 BAU/mL) of anti-RBD-S1 IgG 5 to 6 months after the second vaccine dose (OR 8.2; P = .007, Fisher's exact test). We conclude that although allo-HSCT recipients achieve serum anti-RBD-S1 IgG against SARS-CoV-2 after 2 vaccinations, a deficiency of SARS-CoV-2-specific T-cell immunity may subsequently translate into insufficient humoral responses.
6.
Johansson Wensman, Jonas, et al.
(författare)
The X proteins of bornaviruses interfere with type I interferon signalling
2013
Ingår i: Journal of General Virology. - : Microbiology Society. - 0022-1317 .- 1465-2099. ; 94, s. 263-269
Tidskriftsartikel (refereegranskat) abstract
Borna disease virus (BDV) is a neurotropic, negative-stranded RNA virus causing persistent infection and progressive neurological disorders in a wide range of warm-blooded animals. The role of the small non-structural X protein in viral pathogenesis is not completely understood. Here we investigated whether the X protein of BDV and avian bornavirus (ABV) interferes with the type I interferon (IFN) system, similar to other non-structural proteins of negative-stranded RNA viruses. In luciferase reporter assays, we found that the X protein of various bornaviruses interfered with the type I IFN system at all checkpoints investigated, in contrast to previously reported findings, resulting in reduced type I IFN secretion.
7.
Hansson, Charlotta, et al.
(författare)
Feeding transgenic plants that express a tolerogenic fusion protein effectively protects against arthritis
2016
Ingår i: Plant Biotechnology Journal. - : Wiley-Blackwell. - 1467-7644 .- 1467-7652. ; 14:4, s. 1106-1115
Tidskriftsartikel (refereegranskat) abstract
Although much explored, oral tolerance for treatment of autoimmune diseases still awaits the establishment of novel and effective vectors. We investigated if the tolerogenic CTA1(R7K)-COL-DD fusion protein can be expressed in edible plants and in this way induce oral tolerance and protect against arthritis. The fusion protein was recombinantly expressed in Arabidopsis thaliana plants, which were fed to H-2q restricted DBA/1 mice to assess the preventive effect on collagen-induced arthritis (CIA). The treatment resulted in fewer mice exhibiting disease and arthritis scores were significantly reduced. Immune suppression was evident in treated mice and serum biomarkers for inflammation as well as anti-collagen IgG responses were reduced. In spleen draining and lymph nodes, CD4+ T cell responses were reduced. Concomitant with a reduced effector T cell activity with lower IFNg, IL-13 and IL-17A production we observed an increase in IL-10 production to recall antigen stimulation in vitro, suggesting reduced Th1, Th2 and Th17 activity subsequent to upregulated IL-10 and regulatory T cell (Treg) functions. The present study shows that edible plants expressing a tolerogen were effective at stimulating CD4 T cell tolerance and in protecting against CIA disease. Our study conveys optimism as to the potential of using edible plants for oral treatment of rheumatoid arthritis.
8.
Blodörn, Krister, et al.
(författare)
Vaccine Safety and Efficacy Evaluation of a Recombinant Bovine Respiratory Syncytial Virus (BRSV) with Deletion of the SH Gene and Subunit Vaccines Based On Recombinant Human RSV Proteins: N-nanorings, P and M2-1, in Calves with Maternal Antibodies
2014
Ingår i: PLoS ONE. - : Public Library of Science (PLoS). - 1932-6203. ; 9, s. 1-17
Tidskriftsartikel (refereegranskat) abstract
The development of safe and effective vaccines against both bovine and human respiratory syncytial viruses (BRSV, HRSV) to be used in the presence of RSV-specific maternally-derived antibodies (MDA) remains a high priority in human and veterinary medicine. Herein, we present safety and efficacy results from a virulent BRSV challenge of calves with MDA, which were immunized with one of three vaccine candidates that allow serological differentiation of infected from vaccinated animals (DIVA): an SH gene-deleted recombinant BRSV (Delta SHrBRSV), and two subunit (SU) formulations based on HRSV-P, -M2- 1, and -N recombinant proteins displaying BRSV-F and -G epitopes, adjuvanted by either oil emulsion (Montanide ISA71(VG), SUMont) or immunostimulating complex matrices (AbISCO-300, SUAbis). Whereas all control animals developed severe respiratory disease and shed high levels of virus following BRSV challenge, Delta SHrBRSV-immunized calves demonstrated almost complete clinical and virological protection five weeks after a single intranasal vaccination. Although mucosal vaccination with DSHrBRSV failed to induce a detectable immunological response, there was a rapid and strong anamnestic mucosal BRSV-specific IgA, virus neutralizing antibody and local T cell response following challenge with virulent BRSV. Calves immunized twice intramuscularly, three weeks apart with SUMont were also well protected two weeks after boost. The protection was not as pronounced as that in Delta SHrBRSV-immunized animals, but superior to those immunized twice subcutaneously three weeks apart with SUAbis. Antibody responses induced by the subunit vaccines were non-neutralizing and not directed against BRSV F or G proteins. When formulated as SUMont but not as SUAbis, the HRSV N, P and M2-1 proteins induced strong systemic cross-protective cell-mediated immune responses detectable already after priming. Delta SHrBRSV and SUMont are two promising DIVA-compatible vaccines, apparently inducing protection by different immune responses that were influenced by vaccine-composition, immunization route and regimen.
9.
Nilsson, Elin, et al.
(författare)
Proteomic characterization of IgY preparations purified with a water dilution method.
2008
Ingår i: Journal of agricultural and food chemistry. - : American Chemical Society (ACS). - 1520-5118 .- 0021-8561. ; 56:24, s. 11638-42
Tidskriftsartikel (refereegranskat) abstract
Antigen-specific chicken IgY antibodies have been used for oral immunotherapy as an alternative or complement to antibiotics in several studies. The water dilution (WD) method has several advantages for purifying IgY. It is rapid, efficient, suitable for large-scale production, and nothing but water is added. The water-soluble fraction contains other proteins and lipids besides IgY. The protein content was characterized by two-dimensional gel electrophoresis (2DGE) and nanoflow liquid chromatography coupled offline to matrix-assisted laser desorption/ionization time-of-flight tandem mass spectrometry (nanoLC-MALDI TOF/TOF MS). Protein analysis was complicated due to the large dynamic concentration range, but 26 proteins could be identified. The relative protein concentrations in different batches were very similar according to protein patterns on 1D gels and protein concentration determinations. Thus, the purification method has a high reproducibility. The concentrations of cholesterols and triglycerides were low and should not have an effect on the plasma levels of treated patients. Purification of IgY for oral use with WD is therefore a recommended method.
10.
Johnzon, Carl-Fredrik
(författare)
Inflammatory mechanisms in bacterial infections: focus on mast cells and mastitis
2018
Doktorsavhandling (övrigt vetenskapligt/konstnärligt) abstract
Inflammation is an unspecific response of the immune system to pathogens, for example, invasion by bacteria. This thesis focuses on two aspects of inflammation in the context of bacterial infections: (1) mast cells and (2) mastitis. Mast cells are potent proinflammatory leucocytes that have been implicated in the defence against bacterial infections. Mastitis is an inflammation of the mammary tissue and is one of the most economically destructive disease in the dairy industry worldwide. Here, mast cell synthesis of the potent pro-angiogenic vascular endothelial growth factor (VEGF) in response to stimuli with Staphylococcus aureus (S. aureus) was studied using an in vitro model of primary mouse mast cells. VEGF synthesis was found to be dependent on the presence of live whole bacteria. Previous in vivo investigations of the roles of mast cells in bacterial infections have been conducted using c-Kit-dependent mast cell-deficient mice. These mice suffer from numerous abnormalities in addition to the lack of mast cells. Instead, we used newer, cKit-independent mast cell-deficient mice (Mcpt5-Cre), which have fewer non-mast cell related abnormalities. We found no impact of the mast cell deficiency on the course of intraperitoneal S. aureus infection (e.g., bacterial clearance and cytokine production). We differentiated the virulence of, and response to, a set of clinical bacterial strains of bovine mastitis origin. Escherichia coli (E. coli) and S. aureus strains were injected intraperitoneally into mice. One E. coli strain(strain 127) was found to consistently cause more severe infection (judged by a clinical score) and induce a distinct profile of cytokines (CXCL1, G-CSF, CCL2). The concentrations of these cytokines correlated with both the clinical score and bacterial burden. The kinetics of the clinical and molecular changes that occurred during acute bovine mastitis were studied using a bovine in vivo model in which mastitis was induced by an intramammary infusion of E. coli lipopolysaccharide. Changes in clinical parameters (clinical score, milk changes, rectal temperature) as well as in milk and plasma cytokine concentrations and changes in the metabolome were registered. The progression of these changes occurred in the following order: (1) signs of inflammation in the udder and an increase in milk cytokine concentrations (after/at two hours), (2) visible changes in the milk and an increase in milk somatic cell counts (SCCs) (four hours), (3) changes in the plasma metabolome(four hours) and (4) changes in the milk metabolome (24 hours).
