| 1. |
- Kalbina, Irina, 1961-, et al.
(författare)
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Arabidopsis thaliana plants expressing Rift Valley fever virus antigens : Mice exhibit systemic immune responses as the result of oraladministration of the transgenic plants
- 2016
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Ingår i: Protein Expression and Purification. - San Diego, USA : Elsevier. - 1046-5928 .- 1096-0279. ; 127, s. 61-67
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Tidskriftsartikel (refereegranskat)abstract
- The zoonotic Rift Valley fever virus affects livestock and humans in Africa and on the Arabian Peninsula.The economic impact of this pathogen due to livestock losses, as well as its relevance to public health,underscores the importance of developing effective and easily distributed vaccines. Vaccines that can bedelivered orally are of particular interest.Here, we report the expression in transformed plants (Arabidopsis thaliana) of Rift Valley fever virusantigens. The antigens used in this study were the N protein and a deletion mutant of the Gn glycoprotein.Transformed lines were analysed for specific mRNA and protein content by RT-PCR and Westernblotting, respectively. Furthermore, the plant-expressed antigens were evaluated for their immunogenicityin mice fed the transgenic plants. After oral intake of fresh transgenic plant material, a proportionof the mice elicited specific IgG antibody responses, as compared to the control animals that were fedwild-type plants and of which none sero-converted.Thus, we show that transgenic plants can be readily used to express and produce Rift Valley Fever virusproteins, and that the plants are immunogenic when given orally to mice. These are promising findingsand provide a basis for further studies on edible plant vaccines against the Rift Valley fever virus.
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| 2. |
- Brosché, Mikael, et al.
(författare)
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Gene regulation by low level UV-B radiation : identification by DNA array analysis
- 2002
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Ingår i: Photochemical and Photobiological Sciences. - 1474-905X .- 1474-9092. ; 1:9, s. 656-664
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Tidskriftsartikel (refereegranskat)abstract
- UV-B radiation alters transcript levels of various defence genes and photosynthetic genes in plants. Utilising a DNA array with 5000 ESTs and cDNAs from Arabidopsis thaliana, 70 genes were found to show a greater than two-fold induction or repression of transcript levels. Six genes (MEB5.2, PyroA, Ubq3, Lhcb6, F5D21.10 and the gene for an RNA polymerase II subunit) were tested for stress specific gene regulation on northern blots with RNA from plants exposed to low dose UV-B radiation, ozone or wounding. Transcript levels for PyroA, Uhq3 and the gene for a RNA polymerase II subunit were all specifically increased by UV-B. MEB5.2 mRNA levels also rose, whereas Lhcb6 and FSD21.10 transcript levels decreased under all stresses. The PyroA gene product in fungi is needed for biosynthesis of pyridoxine, and might have a role in protection against singlet oxygen. The Ubq3 gene encodes the ubiquitin protein that is attached to proteins destined for degradation. MEB5.2 and F5D21.10 represent novel gene products whose function have not yet been identified. Pairwise comparisons between the UV-B inducible promoters have identified a series of elements present in the MEB5.2 and PyroA promoters, absent from promoters of genes for early phenylpropanoid metabolism and that may be responsible for modulating their UV-B responses.
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| 3. |
- Castro Alves, Victor, 1986-, et al.
(författare)
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Integration of non-target metabolomics and sensory analysis unravels vegetable plant metabolite signatures associated with sensory quality : A case study using dill (Anethum graveolens)
- 2021
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Ingår i: Food Chemistry. - : Elsevier. - 0308-8146 .- 1873-7072. ; 344
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Tidskriftsartikel (refereegranskat)abstract
- Using dill (Anethum graveolens L.) as a model herb, we revealnovel associations between metabolite profile and sensory quality, by integrating non-target metabolomics with sensory data. Low night temperatures and exposure to UV-enriched light was used to modulate plant metabolism, thereby improving sensory quality. Plant age is a crucial factor associated with accumulation of dill ether and α-phellandrene, volatile compounds associated with dill flavour. However, sensory analysis showed that neither of these compounds has any strong association with dill taste. Rather, amino acids alanine, phenylalanine, glutamic acid, valine, and leucine increased in samples exposed to eustress and were positively associated with dill and sour taste. Increases in amino acids and organic acids changed the taste from lemon/grass to a more bitter/pungent dill-related taste. Our approach reveals a novel approach to establish links between effects of eustressors on sensory quality, and may be applicable to a broad range of crops.
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| 4. |
- Castro Alves, Victor, 1986-, et al.
(författare)
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The taste of UV light : Using sensomics to improve horticultural quality
- 2020
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Ingår i: UV4Plants Bulletin. - Helsingfors : University of Helsinki. - 2343-323X. ; :1, s. 39-43
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Tidskriftsartikel (refereegranskat)abstract
- Greenhouse horticulture is in its broad definition the production of plant products within, under or sheltered by structures that provide protection against biotic and/or abiotic stress. In greenhouses, horticultural crops can grow protected from infectious agents and adverse weather conditions, allowing off-season, year-round production. However, greenhouse production often comes with a trade-off, which is a skewed light environment with a lack of UV light. In some instances, the blockage of UV by greenhouse glass and plastic covers is beneficial from a commercial perspective, especially on tropical latitudes where plants can often encounter higher UV levels, which may impair plant growth and nutrient absorption (Krause et al. 1999; Verdaguer et al. 2017). On the other hand, reduced UV inside greenhouses may reduce the synthesis of metabolites associated with crop protection against biotic and abiotic stress, such as flavonoids, terpenoids and alkaloids (Yang et al. 2018). This reduction in the amount of protective compounds may not be seen as an important limitation in a protected environment, but these metabolic changes caused by reduced UV exposure may in fact negatively impact on product quality. For example, it is possible to improve of the aroma and taste of greenhouse tomato by exposing plants to low levels of supplementary UV light (Dzakovich et al. 2016).
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| 5. |
- Hadad, Ronza, 1984-, et al.
(författare)
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Protection against genital tract Chlamydia trachomatis infection following intranasal immunization with a novel recombinant MOMP VS2/4 antigen
- 2016
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Ingår i: Acta Pathologica, Microbiologica et Immunologica Scandinavica (APMIS). - Hoboken, USA : Wiley-Blackwell. - 0903-4641 .- 1600-0463. ; 124, s. 1078-1086
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Tidskriftsartikel (refereegranskat)abstract
- The asymptomatic nature of most Chlamydia trachomatis infections and the lack of appropriate effects by current prevention and management call for vaccine development. We evaluated a recombinant subunit vaccine candidate based on the major outer membrane protein variable segments 2 and 4 (MOMP VS2/4). To achieve maximal immunogenicity and ease of production and purification, MOMP VS2/4 was constructed by using highly immunogenic sequences of MOMP only, thereby minimizing the presence of hydrophobic regions, and spacing the immunogenic epitopes with a flexible amino acid sequence. A purification tag was also added. The MOMP VS2/4 was given intranasally, with or without intravaginal boost, with cholera toxin (CT) adjuvant to C57BL/6 mice, which were screened for immunogenicity and protection against a live challenge infection with C. trachomatis serovar D. Bacterial shedding, cell-mediated responses, and antibody responses were monitored. Immunized mice exhibited significantly less bacterial shedding and were better protected against infertility as compared to unimmunized control mice. Immunizations stimulated both systemic and local specific antibody (IgG1, IgG2c, and IgA) responses, and primed T cells that produced interferon-c and interleukins 13 and 17 upon challenge with recall antigen. Thus, MOMP VS2/4, in combination with CT adjuvant, stimulated Th1, Th2, and Th17 effector cells, and generated protective immunity associated with less pathology. We regard MOMP VS2/4 as a promising candidate for further development into a mucosal chlamydial vaccine.
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| 6. |
- Hansson, Charlotta, et al.
(författare)
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Feeding transgenic plants that express a tolerogenic fusion protein effectively protects against arthritis
- 2016
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Ingår i: Plant Biotechnology Journal. - : Wiley-Blackwell. - 1467-7644 .- 1467-7652. ; 14:4, s. 1106-1115
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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.
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| 7. |
- Kalbina, Irina, et al.
(författare)
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A novel chimeric MOMP antigen expressed in Escherichia coli, Arabidopsis thaliana, and Daucus carota as a potential Chlamydia trachomatis vaccine candidate
- 2011
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Ingår i: Protein Expression and Purification. - : Elsevier BV. - 1046-5928 .- 1096-0279. ; 80:2, s. 194-202
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Tidskriftsartikel (refereegranskat)abstract
- The major outer membrane protein (MOMP) of Chlamydia trachomatis is a highly antigenic and hydrophobic transmembrane protein. Our attempts to express the full-length protein in a soluble form in Escherichia coli and in transgenic plants failed. A chimeric gene construct of C trachomatis serovar E MOMP was designed in order to increase solubility of the MOMP protein but with retained antigenicity. The designed construct was successfully expressed in E. coil, in Arabidopsis thaliana, and in Daucus carota. The chimeric MOMP expressed in and purified from E. coil was used as antigen for production of antibodies in rabbits. The anti-chimeric MOMP antibodies recognized the corresponding protein in both E. coli and in transgenic plants, as well as in inactivated C. trachomatis elementary bodies. Transgenic Arabidopsis and carrots were characterized for the number of MOMP chimeric genetic inserts and for protein expression. Stable integration of the transgene and the corresponding protein expression were demonstrated in Arabidopsis plants over at least six generations. Transgenic carrots showed a high level of expression of the chimeric MOMP - up to 3% of TSP.
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| 8. |
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| 9. |
- Kalbina, Irina, 1961-, et al.
(författare)
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Construction, immunogenicity and protective efficacy in mice of a prototype chimeric Chlamydia trachomatis MOMP vaccine candidate antigen
- 2011
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Konferensbidrag (refereegranskat)abstract
- A chimeric gene construct of Chlamydia trachomatis serovar E major outer membrane protein (MOMP) was designed, and expressed as a candidate vaccine antigen. The construct was based on known T and B cell epitopes located in the variable segment (VS) 2 and 4 loops of MOMP, and successfully expressed and purified in a recombinant Escherichia coli system. BALB/c mice were immunized intranasally with the chimeric MOMP antigen and Cholera toxin (CT) adjuvant, three immunizations with 10 days intervals. A final boost with the identical antigen preparation was given intravaginally. Challenge with live C. trachomatis serovar D was performed 10 days after boost. Antibodies in serum and vaginal washes were determined with the identical chimeric MOMP construct as antigen in ELISAs. All mice in vaccine groups (N=10/group and experiment) developed a strong antigen-specific IgG response in serum, and some also had detectable antigen-specific IgG in vaginal washes. An IgA response, albeit weaker, was detected in some of the mice both in serum and in vaginal washes.After challenge with C. trachomatis, 80 and 100% of the mice became infected in two experiments, respectively. However, the vaccinated groups cleared the infection significantly faster than control groups (all vaccinated mice healthy day 24 [90% day 16], compared to day 40 for controls).Thus, the new chimeric MOMP antigen construct gave rise to a significant immune response in mice (s-IgG). It also conferred substantial protection to infection caused by genital C. trachomatis infection of a different subtype.
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| 10. |
- Kalbina, Irina, 1961-, et al.
(författare)
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Expression of chimeric Chlamydia trachomatis MOMP protein antigen in Arabidopsis thaliana and Daucus carota
- 2011
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Ingår i: Molecular farming. - Bryssel : COST. ; , s. 38-38
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Konferensbidrag (refereegranskat)abstract
- Urogenital chlamydial infection, caused by Chlamydia trachomatis, is the main sexually transmitted infection in Sweden. Despite active programmes for detection and case finding, nearly 37 000 cases were reported in 2010. Serovar E strains are considered to cause approximately 40-50% of these cases. A vaccine would be highly valuable in order to control the epidemic.The major outer membrane protein (MOMP) of Chlamydia trachomatis is a highly antigenic and hydrophobic transmembrane protein. Our attempts to express the full-length protein in a soluble form in transgenic plants failed. A chimeric gene construct of Chlamydia trachomatis serovar E MOMP was designed in order to increase solubility of the MOMP protein but with retained antigenicity. The construct was based on known T and B cell epitopes located in the variable segment (VS) 2 and 4 loops of MOMP.The designed construct was successfully expressed in Arabidopsis thaliana, and in Daucus carota. A chimeric MOMP expressed in and purified from E. coli was used as antigen for production of antibodies in rabbits. The anti-chimeric MOMP antibodies recognized the corresponding protein in the transgenic plants, as well as in inactivated C. trachomatis elementary bodies. Transgenic Arabidopsis and carrots were characterized for the number of MOMP chimeric genetic inserts and for protein expression. Stable integration of the transgene and the corresponding protein expression were demonstrated in Arabidopsis plants over at least six generations. Transgenic carrots showed a high level of expression of the chimeric MOMP– up to 3% of TSP.
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