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  • 1. Brosché, Mikael
    et al.
    Schuler, Mary A.
    Kalbina, Irina
    Örebro University, Department of Natural Sciences.
    Connor, Lynn
    Strid, Åke
    Örebro University, Department of Natural Sciences.
    Gene regulation by low level UV-B radiation: identification by DNA array analysis2002In: Photochemical and Photobiological Sciences, ISSN 1474-905X, E-ISSN 1474-9092, Vol. 1, no 9, p. 656-664Article in journal (Refereed)
    Abstract [en]

    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.

  • 2.
    Hadad, Ronza
    et al.
    Örebro Life Science Center, School of Science and Technology, Örebro University, Örebro, Sweden; Department of Laboratory Medicine, Faculty of Medicine and Health, Örebro University, Örebro, Sweden.
    Marks, Ellen
    Department of Medical Microbiology and Immunology, University of Gothenburg, Gothenburg, Sweden.
    Kalbina, Irina
    Örebro University, School of Science and Technology. Örebro Life Science Center, Örebro University, Örebro, Sweden.
    Schön, Karin
    Department of Medical Microbiology and Immunology, University of Gothenburg, Gothenburg, Sweden.
    Unemo, Magnus
    Örebro University, School of Medical Sciences. Department of Laboratory Medicine, Örebro University Hospital, Örebro, Sweden.
    Lycke, Nils
    Department of Medical Microbiology and Immunology, University of Gothenburg, Gothenburg, Sweden.
    Strid, Åke
    Örebro University, School of Science and Technology. Örebro Life Science Center, Örebro University, Örebro, Sweden.
    Andersson, Sören
    Örebro University, School of Medical Sciences. Department of Laboratory Medicine, Örebro University Hospital, Örebro, Sweden.
    Protection against genital tract Chlamydia trachomatis infection following intranasal immunization with a novel recombinant MOMP VS2/4 antigen2016In: Acta Pathologica, Microbiologica et Immunologica Scandinavica (APMIS), ISSN 0903-4641, E-ISSN 1600-0463, Vol. 124, p. 1078-1086Article in journal (Refereed)
    Abstract [en]

    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.

  • 3.
    Hansson, Charlotta
    et al.
    Department of Microbiology and Immunology, University of Gothenburg, Gothenburg, Sweden.
    Schön, Karin
    Department of Microbiology and Immunology, University of Gothenburg, Gothenburg, Sweden.
    Kalbina, Irina
    Örebro University, School of Science and Technology.
    Strid, Åke
    Örebro University, School of Science and Technology.
    Andersson, Sören
    Örebro University Hospital. Sch Sci & Technol, Orebro Life Sci Ctr, Univ Orebro, Orebro, Sweden; Department of Laboratory Medicine, Örebro University Hospital, Örebro, Sweden .
    Bokarewa, Maria I.
    Department of Rheumatology and Inflammation Research, University of Gothenburg, Gothenburg, Sweden.
    Lycke, Nils Y.
    Department of Microbiology and Immunology, University of Gothenburg, Gothenburg, Sweden.
    Feeding transgenic plants that express a tolerogenic fusion protein effectively protects against arthritis2016In: Plant Biotechnology Journal, ISSN 1467-7644, E-ISSN 1467-7652, Vol. 14, no 4, p. 1106-1115Article in journal (Refereed)
    Abstract [en]

    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.

  • 4.
    Kalbina, Irina
    Örebro University, Department of Natural Sciences.
    The molecular mechanisms behind perception and signal transduction of UV-B irradiation in Arabidopsis thaliana2005Doctoral thesis, comprehensive summary (Other academic)
    Abstract [en]

    Elevation of UV-B radiation (280-315 nm), occurring as a result of depletion of the stratospheric ozone, causes a number of physiological and biochemical changes in plants. Damage to the photosynthetic apparatus (including the bleaching of the pigments which trap the sun's energy), to the processes of cell division and growth regulation, and to the composition and replication of genetic material are just some of these changes. The consequences include reduction in growth yield, changes in levels and effects of plant hormones and secondary metabolites, and alteration of interactions between plants and other organisms.

    This thesis deals with several mechanistic questions related to regulation of responses during UV-B stress in plants. Our results show significant ecotype-specific variability in UV-B response in the model plant Arabidopsis thaliana. Differences at the molecular level (expression of PR-5 and steady-state concentration of H2O2) resulted in statistically significant differences in biomass, rosette size and leaf area. Therefore, it is of great importance to pay attention to the responses of the background ecotypes when for instance studying mechanisms of responses toward ultraviolet-B radiation in mutants.

    Using a DNA microarray approach, we found a number of novel genes to be differentially expressed under UV-B radiation. Two of the genes (PYROA and MEB5.2) were later used as molecular markers for monitoring of UV-B stress. Promoters of PYROA and MEB5.2 were compared with promoters of genes for the phenylpropanoid pathway. The comparisons indicated only few common elements with the UV-B-regulated promoters of CHS, PAL and CHI. In contrast, the genes identified as being UV-B regulated in this study (MEB5.2, PYROA and UBQ3), completely lacked elements required for the UV-B induction of CHS, indicating that these genes are regulated by different transcription factors. In addition, novel unidentified cis-elements are probably also present upstream of the transcription start.

    Reverse and forward genetics were used for searching novel genes responsive to UV-B and for examination of proposed candidates of the UV-B signal transduction chain. Screening of more than 2000 T-DNA mutants for differential response to UV-B resulted in the identification of a mutant displaying insensitivity to UV-B induced inhibition of hypocotyl growth. By using the corresponding knock-out mutants, the involvement of NADPH oxidase and MAPK phosphatase 1 in UV-B signalling was demonstrated.

    For the plant to be able to respond appropriately to UV-B irradiation, UV-B quanta have to be absorbed. There are indirect evidences for the existence of specific UV-B receptor(s), whereas the receptor itself still remains unknown. By the classical approach of action spectroscopy, we undertook an attempt to identify the absorption spectra of the chromophore(s) sensing UV-B radiation in plants. The investigated molecular markers revealed the presence of two potential chromophores absorbing in the UV-B region and peaking at 280-290 and 300 nm, respectively.

    List of papers
    1. Gene regulation by low level UV-B radiation: identification by DNA array analysis
    Open this publication in new window or tab >>Gene regulation by low level UV-B radiation: identification by DNA array analysis
    Show others...
    2002 (English)In: Photochemical and Photobiological Sciences, ISSN 1474-905X, E-ISSN 1474-9092, Vol. 1, no 9, p. 656-664Article in journal (Refereed) Published
    Abstract [en]

    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.

    Keywords
    Arabidopsis/genetics/radiation effects, Base Sequence, DNA; Plant/genetics, Expressed Sequence Tags, Gene Expression Regulation; Enzymologic/radiation effects, Gene Expression Regulation; Plant/*radiation effects, Molecular Sequence Data, Oligonucleotide Array Sequence Analysis, Plant Proteins/genetics, Promoter Regions (Genetics), RNA Polymerase II/genetics, Ultraviolet Rays
    National Category
    Biochemistry and Molecular Biology
    Research subject
    Biochemistry
    Identifiers
    urn:nbn:se:oru:diva-2879 (URN)10.1039/B202659G (DOI)12665302 (PubMedID)
    Available from: 2005-09-14 Created: 2005-09-14 Last updated: 2017-12-14Bibliographically approved
    2. Supplementary ultraviolet-B irradiation reveals differences in stress responses between Arabidopsis thaliana ecotypes
    Open this publication in new window or tab >>Supplementary ultraviolet-B irradiation reveals differences in stress responses between Arabidopsis thaliana ecotypes
    2005 (English)Manuscript (preprint) (Other academic)
    National Category
    Biochemistry and Molecular Biology
    Research subject
    Biochemistry
    Identifiers
    urn:nbn:se:oru:diva-2880 (URN)
    Available from: 2005-09-14 Created: 2005-09-14 Last updated: 2017-10-18Bibliographically approved
    3. Wavelength dependence of expression of UV-B-induced molecular markers in Arabidopsis thaliana
    Open this publication in new window or tab >>Wavelength dependence of expression of UV-B-induced molecular markers in Arabidopsis thaliana
    (English)Manuscript (Other academic)
    National Category
    Biochemistry and Molecular Biology
    Research subject
    Biochemistry
    Identifiers
    urn:nbn:se:oru:diva-2881 (URN)
    Available from: 2005-09-14 Created: 2005-09-14 Last updated: 2017-10-18Bibliographically approved
    4. The role of NADPH oxidase and MAP kinase phosphatase 1 in UV-B-dependent gene expression in Arabidopsis
    Open this publication in new window or tab >>The role of NADPH oxidase and MAP kinase phosphatase 1 in UV-B-dependent gene expression in Arabidopsis
    (English)Manuscript (Other academic)
    National Category
    Biochemistry and Molecular Biology
    Research subject
    Biochemistry
    Identifiers
    urn:nbn:se:oru:diva-2882 (URN)
    Available from: 2005-09-14 Created: 2005-09-14 Last updated: 2017-10-18Bibliographically approved
    5. An Arabidopsis mutant responsive to UV-B irradiation
    Open this publication in new window or tab >>An Arabidopsis mutant responsive to UV-B irradiation
    (English)Manuscript (Other academic)
    National Category
    Biochemistry and Molecular Biology
    Research subject
    Biochemistry
    Identifiers
    urn:nbn:se:oru:diva-2883 (URN)
    Available from: 2005-09-14 Created: 2005-09-14 Last updated: 2017-10-18Bibliographically approved
  • 5.
    Kalbina, Irina
    et al.
    Örebro University, School of Science and Technology.
    Engstrand, Lars
    Dept Bacteriol, Swedish Inst Infect Dis Control SMI, Solna, Sweden.
    Andersson, Sören
    Örebro University Hospital, Örebro, Sweden; Dept Bacteriol, Swedish Inst Infect Dis Control SMI, Solna, Sweden.
    Strid, Åke
    Örebro University, School of Science and Technology.
    Expression of Helicobacter pylori TonB Protein in Transgenic Arabidopsis thaliana: toward production of vaccine antigens in plants2010In: Helicobacter, ISSN 1083-4389, E-ISSN 1523-5378, Vol. 15, no 5, p. 430-437Article in journal (Refereed)
    Abstract [en]

    Background: The aim of this study was to produce a recombinant version of the highly antigenic Helicobacter pylori TonB (iron-dependent siderophore transporter protein HP1341) in transgenic plants as a candidate oral vaccine antigen. Materials and Methods: Using Agrobacterium-mediated gene transfer, we introduced three different constructs of the tonB gene into the genome of the model plant Arabidopsis thaliana. We investigated transgene insertion by PCR, produced TonB antibodies for analysis of the production of the recombinant protein in plants, verified the identity of the protein produced by mass spectrometry analysis, and analyzed the number of genetic inserts in the plants by Southern blotting. Results: Three different constructs of the expression cassette (full-length tonB, tonB truncated in the 5' end removing the codons for a transmembrane helix, and the latter construct with codons for the endoplasmic reticulum SEKDEL retention signal added to the 3' end) were used to find the most effective way to express the TonB antigen. Production of TonB protein was detected in plants transformed with each of the constructs, confirmed by both Western blotting and mass spectrometry analysis. No considerable differences in protein expression from the three different constructs were observed. The protein concentration in the plants was at least 0.05% of the total soluble proteins. Conclusions: The Helicobacter pylori TonB protein can be produced in Arabidopsis thaliana plants in a form that is recognizable by rabbit anti-TonB antiserum. These TonB-expressing plants are highly suitable for animal studies of oral adminstration as a route for immunization against Helicobacter infections.

  • 6.
    Kalbina, Irina
    et al.
    Örebro University, School of Science and Technology. Orebro Life Science Center.
    Lagerqvist, Nina
    Department of Microbiology, Public Health Agency of Sweden, Solna, Sweden; Department of Microbiology, Tumor and Cell Biology, Karolinska Institutet, Stockholm, Sweden.
    Moiane, Bélisario
    Department of Microbiology, Tumor and Cell Biology, Karolinska Institutet, Stockholm, Sweden; Eduardo Mondlane University, Maputo, Mozambique.
    Ahlm, Clas
    Department of Clinical Microbiology, Umeå University, Umeå, Sweden.
    Andersson, Sören
    Örebro University, School of Medical Sciences. Örebro Life Science Center, Department of Science and Technology, Örebro University, Örebro, Sweden; Department of Clinical Microbiology, Örebro University Hospital, Örebro, Sweden.
    Strid, Åke
    Örebro University, School of Science and Technology.
    Falk, Kerstin I.
    Department of Microbiology, Public Health Agency of Sweden, Solna, Sweden; Department of Microbiology, Tumor and Cell Biology, Karolinska Institutet, Stockholm, Sweden.
    Arabidopsis thaliana plants expressing Rift Valley fever virus antigens: Mice exhibit systemic immune responses as the result of oraladministration of the transgenic plants2016In: Protein Expression and Purification, ISSN 1046-5928, E-ISSN 1096-0279, Vol. 127, p. 61-67Article in journal (Refereed)
    Abstract [en]

    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.

  • 7.
    Kalbina, Irina
    et al.
    Örebro University, Department of Natural Sciences.
    Li, Shaoshan
    Björn, Lars Olof
    Strid, Åke
    Örebro University, Department of Natural Sciences.
    Wavelength dependence of expression of UV-B-induced molecular markers in Arabidopsis thalianaManuscript (Other academic)
  • 8.
    Kalbina, Irina
    et al.
    Örebro University, Department of Natural Sciences.
    Li, Shaoshan
    Örebro University, Department of Natural Sciences.
    Kalbin, Georgi
    Örebro University, Department of Natural Sciences.
    Björn, Lars Olof
    Strid, Åke
    Örebro University, Department of Natural Sciences.
    Two separate UV-B radiation wavelength regions control expression of different molecular markers in Arabidopsis thaliana2008In: Functional Plant Biology, ISSN 1445-4408, E-ISSN 1445-4416, Vol. 35, no 3, p. 222-227Article in journal (Refereed)
    Abstract [en]

    Fluence-response curves were obtained at nine wavelengths in the interval 280-360 nm for mRNA transcripts of four molecular markers induced by ultraviolet-B (UV-B) radiation in Arabidopsis thaliana: CHS (encoding chalcone synthase), PDX1.3 (encoding an enzyme involved in formation of pyridoxine), MEB5.2 (encoding a protein with unknown function but which is strongly up-regulated by UV-B), and LHCB1*3 (encoding a chlorophyll a/b binding protein). Intact Arabidopsis plants were irradiated for 3h using a high intensity deuterium radiation source and narrow bandwith filters (Kalbin et al. 2005, J. Biochem. Biophys. Meth. 65, 1-12) without supplementary PAR. The results obtained suggest the existence of two distinct UV-B signal responses: one sensitive between 300 and 310 nm and the other sensitive around 280-290 nm. Among the investigated molecular markers, CHS and PDX1.3 were regulated through the chromophore absorbing around 300 nm, whereas MEB5.2 and LHCB1*3 were regulated through the chromophore absorbing at 280-290 nm. The results obtained show that at least two signal transduction pathways exist that regulate gene expression as a result of absorption of UV-B radiation in plants.

  • 9.
    Kalbina, Irina
    et al.
    Örebro University, School of Science and Technology.
    Marks, Ellen
    University of Gothenburg, Gothenburg, Sweden.
    Lycke, Nils
    University of Gothenburg, Gothenburg, Sweden.
    Lindh, Ingrid
    Örebro University, School of Science and Technology.
    Unemo, Magnus
    Örebro University Hospital, Örebro, Sweden.
    Strid, Åke
    Örebro University, School of Science and Technology.
    Andersson, Sören
    Örebro University Hospital, Örebro, Sweden.
    Construction, immunogenicity and protective efficacy in mice of a prototype chimeric Chlamydia trachomatis MOMP vaccine candidate antigen2011Conference paper (Refereed)
    Abstract [en]

    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.

  • 10.
    Kalbina, Irina
    et al.
    Örebro University, Department of Natural Sciences.
    Strid, Åke
    Örebro University, Department of Natural Sciences.
    An Arabidopsis mutant responsive to UV-B irradiationManuscript (Other academic)
  • 11.
    Kalbina, Irina
    et al.
    Örebro University, Department of Natural Sciences.
    Strid, Åke
    Örebro University, Department of Natural Sciences.
    Supplementary ultraviolet-B irradiation reveals differences in stress responses between Arabidopsis thaliana ecotypes2005Manuscript (preprint) (Other academic)
  • 12.
    Kalbina, Irina
    et al.
    Örebro University, Department of Natural Sciences.
    Strid, Åke
    Örebro University, Department of Natural Sciences.
    Supplementary ultraviolet-B irradiation reveals differences in stress responses between Arabidopsis thaliana ecotypes2006In: Plant, Cell and Environment, ISSN 0140-7791, E-ISSN 1365-3040, Vol. 29, no 5, p. 754-763Article in journal (Refereed)
    Abstract [en]

    Irradiation of Arabidopsis thaliana ecotypes C24, Wassilewskija (Ws) and Columbia-0 (Col-0) with supplementary ultraviolet-A+B (UV-A+B) radiation revealed ecotype-specific differences in expression of the gene for the pathogenesis-related protein PR-5. C24 showed an increased expression level of PR-5 (5- and 20-fold higher compared with Col-0 and Ws, respectively). Expression of other molecular markers such as CHS (encoding chalcone synthase), MEB5.2 [encoding a gene strongly up-regulated by ultraviolet-B (UV-B)] and PYROA [encoding a pyridoxine (Vitamin B6) biosynthesis enzyme] only showed slight differences between ecotypes. Oxidative stress during UVA+B exposure was monitored by staining for H2O2. This analysis also revealed important ecotype-specific differences. 'H2O2 hot spots' were found in C24, whereas an even distribution of H2O2 was found in Ws and Col-0. Necrotic lesions also appeared on C24 leaves after prolonged UV-B exposure. There was a reverse correlation between the H2O2 steady-state concentration and the PR-5 gene expression; Ws showed the highest level of H2O2 accumulation but the lowest expression level of the PR-5 gene. Furthermore, application of paraquat on the rosettes led to similar PR-5 expression and H2O2 accumulation patterns as were found after UV-A+B irradiation. The observed ecotypic differences were also reflected in a statistically significant UV-B-dependent decrease in biomass, rosette size and leaf area for Ws, but not for C24 and Col-0. Our results show that a significant ecotype-specific genetic variability in general UV-B responses in Arabidopsis exists. Moreover, the signal transduction or gene regulation pathway for PR-5 differs from the other molecular markers used in this study.

  • 13.
    Kalbina, Irina
    et al.
    Örebro University, Department of Natural Sciences.
    Strid, Åke
    Örebro University, Department of Natural Sciences.
    The role of NADPH oxidase and MAP kinase phosphatase 1 in UV-B-dependent gene expression in ArabidopsisManuscript (Other academic)
  • 14.
    Kalbina, Irina
    et al.
    Örebro University, Department of Natural Sciences.
    Strid, Åke
    Örebro University, Department of Natural Sciences.
    The role of NADPH oxidase and MAP kinase phosphatase in UV-B-dependent gene expression in Arabidopsis2006In: Plant, Cell and Environment, ISSN 0140-7791, E-ISSN 1365-3040, Vol. 29, no 9, p. 1783-1793Article in journal (Refereed)
    Abstract [en]

    Plant responses to supplementary UV-B irradiation have been reported to include formation of reactive oxygen species (ROS), hydrogen peroxide, in particular, and regulation by mitogen-activated protein kinase (MAPK) cascades which in turn are fine-tuned by MAPK phosphatases (MKPs). Here we present direct genetic evidence for the involvement of plasma membrane NADPH oxidase, a source of superoxide and hydrogen peroxide in the apoplasts, in UV-B signalling in Arabidopsis thaliana, by analysis of gene expression of the UV-B molecular markers in NADPH oxidase (atrbohD, F and DF) and MAP kinase phosphatase 1 (MKP1) knockout mutants (mkp1). Whereas the NADPH oxidase mutants were affected in UV-B-dependent CHS, PYROA and MEB5.2 gene expression, the mkp1 mutant was affected in the general expression pattern of the pathogenesis-related (PR) and PDF1.2 genes. The results indicate involvement of MKP1 in repressive action on gene expression of more general stress response pathways, similar to those activated by pathogen attack, while NADPH oxidase is involved in quantitative (rather than absolute) regulation of more UV-B-specific genes. The expressions of the molecular markers in the knockout mutant mkp1 and in its complemented lines (lines 6 and 10) were similar, as opposed to the responses of the corresponding wild-type Wassilewskija-4 (Ws-4). Lines 6 and 10 showed much higher MKP1 mRNA than Ws-4 but did not complement the mutant. This suggests a complex dependency of the MAPK phosporylation level of the PR and PDF1.2 genes. Both NADPH oxidase mutants and the mkp1 mutant phenotypically responded to UV-B by growth retardation.

  • 15.
    Kalbina, Irina
    et al.
    Örebro University, School of Science and Technology.
    Wallin, Anita
    Uppsala universitet, Uppsala, Sweden.
    Lindh, Ingrid
    Örebro University, School of Science and Technology.
    Engström, Peter
    Uppsala universitet, Uppsala, Sweden.
    Andersson, Sören
    Örebro University Hospital, Örebro, Sweden.
    Strid, Åke
    Örebro University, School of Science and Technology.
    A novel chimeric MOMP antigen expressed in Escherichia coli, Arabidopsis thaliana, and Daucus carota as a potential Chlamydia trachomatis vaccine candidate2011In: Protein Expression and Purification, ISSN 1046-5928, E-ISSN 1096-0279, Vol. 80, no 2, p. 194-202Article in journal (Refereed)
    Abstract [en]

    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. coli, in Arabidopsis thaliana, and in Daucus carota. The 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 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.

  • 16.
    Kalbina, Irina
    et al.
    Örebro University, School of Science and Technology.
    Wallin, Anita
    Uppsala universitet, Uppsala, Sweden.
    Lindh, Ingrid
    Örebro University, School of Science and Technology.
    Engström, Peter
    Uppsala universitet, Uppsala, Sweden.
    Andersson, Sören
    Universitetssjukhuset i Örebro, Örebro, Sweden.
    Strid, Åke
    Örebro University, School of Science and Technology.
    Expression of chimeric Chlamydia trachomatis MOMP protein antigen in Arabidopsis thaliana and Daucus carota2011In: Molecular farming: plants as a production platform for high value proteins / [ed] Ann Depicker, Bryssel: COST , 2011, p. 38-38Conference paper (Refereed)
    Abstract [en]

    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.

  • 17.
    Lindh, Ingrid
    et al.
    Örebro University, School of Science and Technology. Örebro Life Sci Ctr, Univ Örebro, Örebro, Sweden; Sch Sci & Technol, Univ Örebro, Örebro, Sweden.
    Bråve, Andreas
    Swedish Institute for Communicable Disease Control (SMI), Stockholm, Sweden.
    Hallengärd, David
    Dept Microbiol Tumor & Cell Biol, Karolinska Inst, Stockholm, Sweden.
    Hadad, Ronza
    Örebro University, School of Science and Technology. Örebro Life Sci Ctr, Univ Örebro, Örebro, Sweden.
    Kalbina, Irina
    Örebro University, School of Science and Technology. Örebro Life Sci Ctr, Univ Örebro, Örebro, Sweden.
    Strid, Åke
    Örebro University, School of Science and Technology. Örebro Life Sci Ctr, Univ Örebro, Örebro, Sweden.
    Andersson, Sören
    Örebro University Hospital. Örebro Life Sci Ctr, Univ Örebro, Örebro, Sweden; Örebro University Hospital, Örebro, Sweden.
    Oral delivery of plant-derived HIV-1 p24 antigen in low doses shows a superior priming effect in mice compared to high doses2014In: Vaccine, ISSN 0264-410X, E-ISSN 1873-2518, Vol. 32, no 20, p. 2288-2293Article in journal (Refereed)
    Abstract [en]

    During early infection with human immunodeficiency virus type 1 (HIV-1), there is a rapid depletion of CD4+ T-cells in the gut-associated lymphoid tissue (GALT) in the gastrointestinal tract. Therefore, immediate protection at these surfaces is of high priority for the development of an HIV-1 vaccine. Thus, transgenic plants expressing HIV-1 antigens, which are exposed to immune competent cells in the GALT during oral administration, can be interesting as potential vaccine candidates. In the present study, we used two HIV-1 p24 antigen-expressing transgenic plant systems, Arabidopsis thaliana and Daucus carota, in oral immunization experiments. Both transgenic plant systems showed a priming effect in mice and induced humoral immune responses, which could be detected as anti-p24-specific IgG in sera after an intramuscular p24 protein boost. Dose-dependent antigen analyses using transgenic Arabidopsis thaliana indicated that low p24 antigen doses were superior to high p24 antigen doses

  • 18.
    Lindh, Ingrid
    et al.
    Örebro University, School of Science and Technology.
    Kalbina, Irina
    Örebro University, School of Science and Technology.
    Hedberg, Sara Thulin
    Scherbak, Nikolai
    Örebro University, School of Science and Technology.
    Sävenstrand, Helena
    Örebro University, School of Science and Technology.
    Bråve, Andreas
    Hinkula, Jorma
    Strid, Åke
    Örebro University, School of Science and Technology.
    Andersson, Sören
    Örebro University, School of Science and Technology.
    Feeding of mice with Arabidopsis thaliana expressing the HIV-1 subtype C p24 antigen gives rise to systemic immune responses2008In: Acta Pathologica, Microbiologica et Immunologica Scandinavica (APMIS), ISSN 0903-4641, E-ISSN 1600-0463, Vol. 116, no 11, p. 985-994Article in journal (Refereed)
    Abstract [en]

    Development of transgenic edible plants, to be used as production, storage and delivery systems for recombinant vaccine antigens, is a promising strategy to obtain cost effective vaccines against infectious diseases, not the least for use in developing countries. Therefore, we used Agrobacterium tumefaciens-mediated gene transfer to introduce the p24 gag gene encoding the nucleocapsid protein from HIV-1 subtype C into the Arabidopsis thaliana plant genome. Eighteen plant lines were confirmed positive for the p24 gene by PCR, four of these lines showed an apparent homozygous phenotype when grown on selective medium and these lines also showed transcription of the p24 gene into its corresponding mRNA. The mRNA in all four cases generated the p24 protein in plants, as verified by western blot analysis. The plants were shown to contain between 0.2 µg and 0.5 µg p24 protein per g of fresh tissue. Analysis of the localisation of the p24 protein showed that stem tissue contained the largest amount of protein, more than twice as much as leaf tissue, whereas no p24 protein was detected in roots. By using Southern blotting, we found that 4, 2-3, 2 and 1 T-DNA insertion events took place in the four lines 1, 2, 7, and 10, respectively. The genetic insertions of line 1 were stable from the T1 to the T4 generation and gave rise to the p24 protein in all cases, as verified by western blotting. In mice fed with fresh transgenic A. thaliana (line 10), anti-gag IgG was obtained in serum after a booster injection with recombinant p37Gag. No immune response was observed after equal booster injection of untreated mice or mice fed with A. thaliana WT plants.

  • 19.
    Lindh, Ingrid
    et al.
    Örebro University, School of Science and Technology.
    Wallin, Anita
    Evolutionsbiologiskt Centrum, Uppsala universitet.
    Kalbina, Irina
    Örebro University, School of Science and Technology.
    Sävenstrand, Helena
    Örebro University, School of Science and Technology.
    Engström, Peter
    Uppsala Universitet.
    Andersson, Sören
    Smittskyddsinstitutet.
    Strid, Åke
    Örebro University, School of Science and Technology.
    Production of the p24 capsid protein from HIV-1 subtype C in Arabidopsis thaliana and Daucus carota using an endoplasmic reticulum-directing SEKDEL sequence in protein expression constructs2009In: Protein Expression and Purification, ISSN 1046-5928, E-ISSN 1096-0279, Vol. 66, no 1, p. 46-51Article in journal (Refereed)
    Abstract [en]

    An optimized gene expression construct was designed in order to increase the accumulation of the HIV-1 subtype C p24 protein in Arabidopsis thaliana and carrot (Daucus carota) plants. An ER retention signal was introduced into the genetic construct generating a p24 protein containing a SEKDEL amino acid sequence at its C-terminus. Mature A. thaliana plants and carrot cells were transformed using Agrobacterium tumefaciens carrying the improved pGreen0229/p24_SEKDEL vector. Several transgenic plant lines were obtained from both plant species by growth on selective medium and confirmed by PCR. Transformed lines were analyzed for p24 protein content by western blotting using anti-p24-specific antibodies and by Southern blotting to establish the number of copies of the insert in the plant nuclear genome. To estimate the accumulation levels of p24 protein in the plants, ELISA was run using soluble plant extracts. By comparing these results with our previous findings, the ER retention signal increased the level of p24 protein 5-fold in the Arabidopsis thaliana plants. In carrot taproot, the content of p24_SEKDEL protein was approximately half of that in Arabidopsis on a fresh weight basis and was stable in planta for several months. However, on a total soluble protein basis, carrots produced considerable higher levels of the p24_SEKDEL protein than Arabidopsis.

  • 20.
    Qian, Minjie
    et al.
    Örebro University, School of Science and Technology. Department of Horticulture, The State Agricultural Ministry Key Laboratory of Horticultural Plant Growth, Development & Quality Improvement, Zhejiang University, Hangzhou, Zhejiang Province, China.
    Kalbina, Irina
    Örebro University, School of Science and Technology.
    Rosenqvist, Eva
    Section of Crop Sciences, Department of Plant and Environmental Sciences, Copenhagen University, University of Copenhagen, Copenhagen, Denmark.
    Jansen, Marcel A. K.
    School of Biological, Earth and Environmental Sciences, University College Cork, Cork, Ireland.
    Teng, Yuanwen
    Department of Horticulture, The State Agricultural Ministry Key Laboratory of Horticultural Plant Growth, Development & Quality Improvement, Zhejiang University, Hangzhou, Zhejiang Province, China.
    Strid, Åke
    Örebro University, School of Science and Technology.
    UV regulates expression of phenylpropanoid biosynthesis genes in cucumber (Cucumis sativus L.) in an organ and spectrum dependent manner2019In: Photochemical and Photobiological Sciences, ISSN 1474-905X, E-ISSN 1474-9092, Vol. 18, no 2, p. 424-433Article in journal (Refereed)
    Abstract [en]

    Expression of cucumber (Cucumis sativus) genes encoding the phenylpropanoid and flavonoid biosynthetic enzymes phenylalanine ammonia lyase (PAL), cinnamic acid 4-hydroxylase (C4H), and chalcone synthase (CHS), was studied under control light conditions (photosynthetically active radiation, PAR) in root, stem, and leaf. Furthermore, expression was quantified in leaves illuminated with PAR and supplemental ultraviolet-A (315-400nm) or ultraviolet-B (280-315 nm) radiation. The expression pattern of all twelve CsPAL, threeCsC4H, and three CsCHS genes was established. Among the genes regulated by UV two general expression patterns emerge. One pattern applies to genes primarily regulated by enriched UV-A illumination (pattern 1). Another (pattern 2) was found for the genes regulated by enriched UV-B. Three of the pattern 2 genes (CsPAL4, CsPAL10, CsCHS2) displayed a particular sub-pattern (pattern 2b) with transcription enriched by at least 30 fold. In contrast to the other genes studied, the promoters of the genes regulated according to pattern 2b contained a combination of a number of cis-acting regulatory elements (MREs, ACEs, and G-boxes) that may be of importance for the particularly high enhancement of expression under UV-B- containing light. The regulation of phenylpropanoid and flavonoid biosynthesis genes in cucumber resembles that of a number of other plants. However, cucumber, due to its greater size, is an attractive species for more detailed studies of the fine regulation of spatial and temporal expression of key genes. This in turn, can facilitate the quantitative investigation of the relationships between different promotor motifs, the expression levels of each of these three genes, and metabolite accumulation profiles.

  • 21.
    Qian, Minjie
    et al.
    Örebro Life Science Center, School of Science and Technology, Örebro University, Örebro, Sweden.
    Rosenqvist, Eva
    Section of Crop Sciences, Department of Plant and Environmental Sciences, University of Copenhagen, Taastrup, Sweden.
    Flygare, Ann-Marie
    Örebro University, Örebro University School of Business.
    Kalbina, Irina
    Örebro University, School of Science and Technology.
    Teng, Yuanwen
    Department of Horticulture, The State Agricultural Ministry Key Laboratory of Horticultural Plant Growth, Development & Quality Improvement, Zhejiang University, Hangzhou, , Zhejiang Province, China .
    Jansen, Marcel A. K.
    School of Biological, Earth and Environmental Sciences, Environmental Research Institute, University College Cork, North Mall, Cork, Ireland .
    Strid, Åke
    Örebro University, School of Science and Technology.
    UV-A light induces a robust and dwarfed phenotype in cucumber plants (Cucumis sativus L.) without affecting fruit yield2020In: Scientia Horticulturae, ISSN 0304-4238, E-ISSN 1879-1018, Vol. 263, article id 109110Article in journal (Refereed)
    Abstract [en]

    Solar ultraviolet (UV) light influences plant growth and metabolism. Whereas high doses of UV can be deleterious for plants, natural UV doses are important for morphogenesis in many plants species, including those used in horticulture. Greenhouses are widely used for horticultural production and common cladding materials strongly absorb UV. Thus, low amounts of UV may be limiting the optimal development in some plant species. Light supplementation using UV tubes can overcome UV deficiency. Here we study cucumber seedling production in the absence or presence of different UV wavelengths. UV-A- (315-400 nm) and UV-B- (280-315 nm) enriched light was used for exposure and parameters such as the maximum quantum yield of photosystem II, stem development (internode length and diameter, stem dry weight, stem weight per unit of stem length, and stem bending), root biomass, leaf biomass and specific leaf mass were measured. We found that UV-A supplementation resulted in shorter more compact and sturdy plants, properties that are positive from a horticultural perspective. In contrast, UV-B-enriched light led to even smaller plants that lacked the sturdy phenotype. There were no signs of decreased Fv/Fmunder any of the treatments, nor statistically significant differences in fruit yield between the control plants and the UV-treated plants when grown to harvest. In particular, the differences in fruit yield between the controls and the UV-A-treated plants were negligible in all cases. Thus, supplementary UV-A light can be an interesting alternative to chemical growth regulators for production of sturdy horticultural plants.

  • 22.
    Ristilä, Mikael
    et al.
    Örebro University, Department of Natural Sciences.
    Kalbina, Irina
    Örebro University, Department of Natural Sciences.
    Strid, Åke
    Örebro University, Department of Natural Sciences.
    The role of heterotrimeric G-proteins in ultraviolet-B-stress in Arabidopsis thalianaManuscript (Other academic)
  • 23.
    Strid, Åke
    et al.
    Örebro University, School of Science and Technology.
    Lagerqvist, Nina
    The Public Health Agency of Sweden, Stockholm, Sweden.
    Moiane, B.
    Eduardo Mondlane University, Maputo, Mozambique.
    Kalbina, Irina
    Örebro University, School of Science and Technology.
    Andersson, Sören
    Örebro University Hospital, Örebro, Sweden.
    Falk, Kerstin I.
    The Public Health Agency of Sweden, Stockholm, Sweden.
    Expression of Rift Valley Fever virus antigens in Arabidopsis thaliana for oral consumption2012In: Molecular farming: plants as a production platform for high value proteins, Bryssel: COST , 2012Conference paper (Refereed)
    Abstract [en]

    Rift Valley Fever (RVF) is a viral disease affecting both domesticated ruminants and humans. Since 1931, when the causative agent was first discovered in Kenya [1], there have been several severe outbreaks mostly in Sub-Saharan Africa [2]. RVF is now considered as one of Africa’s most important viral zoonoses and is endemic in large parts of the continent. In recent years, RVF has also emerged into Saudi Arabia and the Yemen, where it now is endemic [3]. Common symptoms of an ongoing RVF infection in humans are influenza-like, although more severe clinical manifestations such as hemorrhagic fever, ocular disease and encephalitis are often observed [4]. Outbreaks in livestock may have large economic impact.

     

    The etiological agent, the RVF virus (RVFV), is an enveloped negative sense RNA virus, which belongs to the genus Phlebovirus in the Bunyaviridae family. As the other members of this family, RVFV has three gene segments; the L, M, and S segments. The L segment encodes an RNA-dependent RNA polymerase and the M-segment the glycoproteins and a non-structural protein. By using an ambisense strategy, the S-segment codes for the highly immunogenic nucleocapsid protein (N) and a non-structural protein [4].

     

    The main focus of this project is to establish the plant production of an RVF vaccine candidate, primarily for oral administration. This is an attractive model for vaccination, especially of livestock. The two currently available vaccines for animals are a live attenuated variant, albeit teratogenic, or a weaker inactivated vaccine which requires annual boosters. There is no human vaccine available for general use.

     

    Similarly to our previous expression studies with the HIV p24 protein [5-7], the Helicobacter pylori TonB protein [8], and the Chlamydia trachomatis MOMP chimera [9], we have used Agrobacterium tumefaciens-mediated gene transfer to introduce genes encoding RVFV antigens into Arabidopsis thaliana. Transformed model plants have been created that express the full length RVFV N protein or deletion mutants of the two RVF glycoproteins. Analyses of transformants are on-going (PCR for genomic insertion, cDNA synthesis and RT-PCR for mRNA occurrence, and Western blotting for protein production) and in at least some cases have been shown to carry the corresponding recombinant protein. Mice are being fed fresh transgenic A. thaliana and the subsequent immune response towards the N protein and the glycoproteins will be closely monitored and evaluated by neutralisation test, Western blot and ELISA. Thereafter, the mice will be challenged with the wild-type virus and the protective efficacy of the edible vaccine will be determined.

     

    References

    1. Daubney R, Garnham P (1931) J Patol Bacterio 34: 8922-8926; 2. Gerdes G (2004) Rev Sci Tech 23: 613-623; 3. Balkhy H, Memish Z (2003) Int J Antimicrob Agents 21: 153-157; 4. Flick R, Bouloy M (2005) Curr Mol Med 5: 827-834; 5. Lindh, I., Kalbina, I., Thulin, S., Scherbak, N., Sävenstrand, H., Bråve, A., Hinkula, J., Strid, Å. & Andersson, S. (2008) APMIS 116, 985-994; 6. Lindh, I., Wallin, A., Kalbina, I., Sävenstrand, H., Engström, P., Andersson, S. & Strid, Å. (2009) Prot. Expr. Purif. 66, 46-51; 7. Lindh, I., Andersson, S. & Strid, Å. (2010) In vivo 24, 368-370; 8. Kalbina, I., Engstrand, L., Andersson, S. & Strid, Å. (2010) Helicobacter 15, 430-437; 9. Kalbina I., Wallin A., Lindh I., Engström P., Andersson S. & Strid Å. (2011) Prot. Expr. Purif. 80, 194-202.

  • 24.
    Strid, Åke
    et al.
    Örebro University, School of Science and Technology.
    Lindh, Ingrid
    Örebro University, School of Science and Technology.
    Kalbina, Irina
    Örebro University, School of Science and Technology.
    Andersson, Sören
    Örebro University Hospital, Örebro, Sweden.
    Plant-based production of vaccine antigens and adjuvants2010Conference paper (Refereed)
    Abstract [en]

    Vaccination is the most successful form of prevention of infectious diseases but only a few dozen of several hundred infectious diseases can be prevented in this way due to the extreme cost of vaccine development and due to costly production. Obviously, there is a growing need for large scale and low cost production of valuable proteins, such as vaccine antigens, and alternative methods to mammalian and microbial production systems are also desirable. Plants/plant cells may offer such an option. In our project, we use plants for synthesis of vaccine antigens and adjuvant proteins. Among targeted diseases are HIV, Chlamydia trachomatis (CT) and Helicobacter pylori infections. Novel genetic constructs have been designed for production of optimized antigen proteins and several transformation techniques and intracellular protein production sites are being examined for yield optimization. Both Arabidopsis and carrot have been succesfully used as production hosts and plant-produced antigens against HIV and CT have been used in immunization trials in laboratory mice, giving rise to systemic immune responses. For CT, our designed chimeric protein has also been shown to protect against the disease. Moreover, to further increase the effect of vaccine antigens, we also use plants to produce protein-based adjuvants for inclusion in vaccine formulations. This project is part of the transnational COST network FA0804: ‘Molecular farming - plants as production platform for high-value proteins’. A few slides will be devoted to the brief presentation of the collaboration within this network, which is comprised of research groups in 23 EU countries and a few associated groups outside EU.

  • 25.
    Strid, Åke
    et al.
    Örebro University, School of Science and Technology.
    Lindh, Ingrid
    Örebro University, School of Science and Technology.
    Kalbina, Irina
    Örebro University, School of Science and Technology.
    Engstrand, Lars
    Smittskyddsinstitutet, The Public Health Agency, Solna, Sweden.
    Lycke, Nils
    Gothenburg University, Gothenburg, Sweden.
    Marks, Ellen
    Gothenburg University, Gothenburg, Sweden.
    Applequist, Steve
    Karolinska Institute, Stockholm, Sweden.
    Andersson, Sören
    Örebro University Hospital, Örebro, Sweden.
    HIV, Chlamydia trachomatis and Helicobacter pylori vaccine antigen and proteinaceous adjuvant production in arabidopsis and carrot2010In: Molecular farming: plant as a production platform for high value proteins / [ed] Nunzia Scotti, Bryssel: COST , 2010, p. 17-17Conference paper (Refereed)
    Abstract [en]

    In our project, we use plants for synthesis of vaccine antigens and adjuvant proteins. Among targeted diseases are HIV, Chlamydia trachomatis (CT) and Helicobacter pylori (HP) infections. Novel genetic constructs have been designed for production of optimized antigen proteins and several transformation techniques and intracellular protein production sites are being examined for yield optimization. Both Arabidopsis thaliana (HIV, CT, HP) and carrot (HIV, CT) have been successfully used as production hosts and plant-produced antigens against HIV (primarily the p24 protein), CT (own designed chimera of the MOMP protein) and HP (different versions of the TonB protein). For HIV and CT these antigens have been used in immunization trials in laboratory mice, giving rise to systemic immune responses. For this purpose both consumption of plant tissue and distribution of purified antigens have been used. For CT, the administration of the recombinant protein has also been shown to protect against the disease in mice. Moreover, to further increase the effect of vaccine antigens, we also use plants to produce protein-based adjuvants for inclusion in vaccine formulations. These adjuvants are based either on the bacterial flagellin protein or are cholera toxin-derived chimeric proteins.

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