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Kalbina, Irina
Publications (10 of 25) Show all publications
Qian, M., Rosenqvist, E., Flygare, A.-M., Kalbina, I., Teng, Y., Jansen, M. A. K. & Strid, Å. (2020). UV-A light induces a robust and dwarfed phenotype in cucumber plants (Cucumis sativus L.) without affecting fruit yield. Scientia Horticulturae, 263, Article ID 109110.
Open this publication in new window or tab >>UV-A light induces a robust and dwarfed phenotype in cucumber plants (Cucumis sativus L.) without affecting fruit yield
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2020 (English)In: Scientia Horticulturae, ISSN 0304-4238, E-ISSN 1879-1018, Vol. 263, article id 109110Article in journal (Refereed) Published
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.

Place, publisher, year, edition, pages
Elsevier, 2020
Keywords
Cucumber production, Fruit yield, Plant architecture, Plant morphology, Ultraviolet light
National Category
Plant Biotechnology Horticulture Botany Biochemistry and Molecular Biology
Research subject
Biochemistry
Identifiers
urn:nbn:se:oru:diva-78473 (URN)10.1016/j.scienta.2019.109110 (DOI)000508491600018 ()2-s2.0-85076304828 (Scopus ID)
Funder
Knowledge Foundation, 20130164Swedish Research Council Formas, 942-2015-516
Note

Funding Agencies:

Faculty for Business, Science and Technology at Örebro University

Science Foundation Ireland S16/IA/4418

China Scholarship Council 201406320076

Available from: 2019-12-05 Created: 2019-12-05 Last updated: 2020-03-17Bibliographically approved
Qian, M., Kalbina, I., Rosenqvist, E., Jansen, M. A. K., Teng, Y. & Strid, Å. (2019). UV regulates expression of phenylpropanoid biosynthesis genes in cucumber (Cucumis sativus L.) in an organ and spectrum dependent manner. Photochemical and Photobiological Sciences, 18(2), 424-433
Open this publication in new window or tab >>UV regulates expression of phenylpropanoid biosynthesis genes in cucumber (Cucumis sativus L.) in an organ and spectrum dependent manner
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2019 (English)In: Photochemical and Photobiological Sciences, ISSN 1474-905X, E-ISSN 1474-9092, Vol. 18, no 2, p. 424-433Article in journal (Refereed) Published
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.

Place, publisher, year, edition, pages
London, UK: Royal Society of Chemistry, 2019
National Category
Biochemistry and Molecular Biology Botany
Research subject
Biochemistry
Identifiers
urn:nbn:se:oru:diva-70117 (URN)10.1039/C8PP00480C (DOI)000458569100015 ()30628617 (PubMedID)2-s2.0-85061384256 (Scopus ID)
Funder
Swedish Research Council Formas, 942-2015-516Knowledge Foundation, 20130164The Swedish Foundation for International Cooperation in Research and Higher Education (STINT), CH2016-6788
Note

Funding Agencies:

Orebro University's Faculty for Business, Science and Technology  

Science Foundation Ireland  SFI 16-IA-4418 

National Natural Science Foundation of China  31711530027 

China Scholarship Council (CSC) 

Available from: 2018-11-11 Created: 2018-11-11 Last updated: 2019-06-18Bibliographically approved
Kalbina, I., Lagerqvist, N., Moiane, B., Ahlm, C., Andersson, S., Strid, Å. & Falk, K. I. (2016). Arabidopsis thaliana plants expressing Rift Valley fever virus antigens: Mice exhibit systemic immune responses as the result of oraladministration of the transgenic plants. Protein Expression and Purification, 127, 61-67
Open this publication in new window or tab >>Arabidopsis thaliana plants expressing Rift Valley fever virus antigens: Mice exhibit systemic immune responses as the result of oraladministration of the transgenic plants
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2016 (English)In: Protein Expression and Purification, ISSN 1046-5928, E-ISSN 1096-0279, Vol. 127, p. 61-67Article in journal (Refereed) Published
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.

Place, publisher, year, edition, pages
San Diego, USA: Elsevier, 2016
Keywords
Antigen production, Arabidopsis thaliana, Rift valley fever virus, Plant vaccine, Transformation
National Category
Immunology in the medical area Immunology Biochemistry and Molecular Biology Medical Biotechnology (with a focus on Cell Biology (including Stem Cell Biology), Molecular Biology, Microbiology, Biochemistry or Biopharmacy)
Research subject
Biochemistry; Immunology
Identifiers
urn:nbn:se:oru:diva-51367 (URN)10.1016/j.pep.2016.07.003 (DOI)000382181300009 ()27402440 (PubMedID)2-s2.0-84978634507 (Scopus ID)
Projects
Vaccinutveckling och vaccinproduktion
Funder
Knowledge FoundationStiftelsen Olle Engkvist Byggmästare
Note

Funding Agencies:

Swedish International Development Cooperation Agency (SIDA)

Örebro University's Faculty for Business, Science and Technology

Sparbanksstiftelsen Nya

Available from: 2016-07-17 Created: 2016-07-17 Last updated: 2018-01-10Bibliographically approved
Hansson, C., Schön, K., Kalbina, I., Strid, Å., Andersson, S., Bokarewa, M. I. & Lycke, N. Y. (2016). Feeding transgenic plants that express a tolerogenic fusion protein effectively protects against arthritis. Plant Biotechnology Journal, 14(4), 1106-1115
Open this publication in new window or tab >>Feeding transgenic plants that express a tolerogenic fusion protein effectively protects against arthritis
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2016 (English)In: Plant Biotechnology Journal, ISSN 1467-7644, E-ISSN 1467-7652, Vol. 14, no 4, p. 1106-1115Article in journal (Refereed) Published
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.

Place, publisher, year, edition, pages
Wiley-Blackwell, 2016
Keywords
autoimmunity; transgenic plants; edible plants; CIA; IL-10; FoxP3
National Category
Immunology in the medical area Plant Biotechnology Biochemistry and Molecular Biology
Research subject
Biochemistry; Immunology; Molecular Biology
Identifiers
urn:nbn:se:oru:diva-45625 (URN)10.1111/pbi.12479 (DOI)000373069400006 ()26403330 (PubMedID)2-s2.0-84961215734 (Scopus ID)
Funder
Swedish Cancer SocietySwedish Research CouncilKnut and Alice Wallenberg FoundationAFA InsuranceEU, FP7, Seventh Framework ProgrammeSwedish Foundation for Strategic Research
Note

Funding Agency:

UNISEC

Available from: 2015-08-22 Created: 2015-08-22 Last updated: 2018-08-31Bibliographically approved
Hadad, R., Marks, E., Kalbina, I., Schön, K., Unemo, M., Lycke, N., . . . Andersson, S. (2016). Protection against genital tract Chlamydia trachomatis infection following intranasal immunization with a novel recombinant MOMP VS2/4 antigen. Acta Pathologica, Microbiologica et Immunologica Scandinavica (APMIS), 124, 1078-1086
Open this publication in new window or tab >>Protection against genital tract Chlamydia trachomatis infection following intranasal immunization with a novel recombinant MOMP VS2/4 antigen
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2016 (English)In: Acta Pathologica, Microbiologica et Immunologica Scandinavica (APMIS), ISSN 0903-4641, E-ISSN 1600-0463, Vol. 124, p. 1078-1086Article in journal (Refereed) Published
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.

Place, publisher, year, edition, pages
Hoboken, USA: Wiley-Blackwell, 2016
Keywords
Chlamydia trachomatis, vaccine, major outer membrane protein, mice, antibody response, T cells
National Category
Immunology in the medical area Microbiology in the medical area
Research subject
Biochemistry; Immunology; Microbiology; Infectious Diseases
Identifiers
urn:nbn:se:oru:diva-53554 (URN)10.1111/apm.12605 (DOI)000388265700008 ()27859689 (PubMedID)2-s2.0-84995753108 (Scopus ID)
Projects
Utveckling av vacciner mot sexuellt överförbara sjukdomarMolecular farming
Funder
Stiftelsen Olle Engkvist Byggmästare
Note

Funding Agencies:

Sparbanksstiftelsen Nya

Örebro University's Faculty for Business, Science, and Technology

Foundation for Medical Research at Örebro University Hospital

Available from: 2016-11-18 Created: 2016-11-18 Last updated: 2019-03-06Bibliographically approved
Lindh, I., Bråve, A., Hallengärd, D., Hadad, R., Kalbina, I., Strid, Å. & Andersson, S. (2014). Oral delivery of plant-derived HIV-1 p24 antigen in low doses shows a superior priming effect in mice compared to high doses. Vaccine, 32(20), 2288-2293
Open this publication in new window or tab >>Oral delivery of plant-derived HIV-1 p24 antigen in low doses shows a superior priming effect in mice compared to high doses
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2014 (English)In: Vaccine, ISSN 0264-410X, E-ISSN 1873-2518, Vol. 32, no 20, p. 2288-2293Article in journal (Refereed) Published
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

Place, publisher, year, edition, pages
Elsevier, 2014
Keywords
Transgenic plants; Arabidopsis thaliana; Carrot; Edible vaccine; Oral immunization; HIV-1 p24
National Category
Immunology in the medical area Medical Biotechnology (with a focus on Cell Biology (including Stem Cell Biology), Molecular Biology, Microbiology, Biochemistry or Biopharmacy) Biochemistry and Molecular Biology
Research subject
Biochemistry; Immunology
Identifiers
urn:nbn:se:oru:diva-33982 (URN)10.1016/j.vaccine.2014.02.073 (DOI)000335100600005 ()24631072 (PubMedID)2-s2.0-84897502968 (Scopus ID)
Projects
Ätbara vacciner
Funder
Knowledge Foundation
Note

Funding Agencies:

Örebro University's Faculty for Business, Science, and Technology 

Nyckelfonden

Available from: 2014-02-27 Created: 2014-02-27 Last updated: 2018-06-05Bibliographically approved
Strid, Å., Lagerqvist, N., Moiane, B., Kalbina, I., Andersson, S. & Falk, K. I. (2012). Expression of Rift Valley Fever virus antigens in Arabidopsis thaliana for oral consumption. In: Molecular farming: plants as a production platform for high value proteins. Paper presented at COST FA0804, Warszawa 5-7th September 2012. Bryssel: COST
Open this publication in new window or tab >>Expression of Rift Valley Fever virus antigens in Arabidopsis thaliana for oral consumption
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2012 (English)In: Molecular farming: plants as a production platform for high value proteins, Bryssel: COST , 2012Conference paper, Oral presentation with published abstract (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.

Place, publisher, year, edition, pages
Bryssel: COST, 2012
National Category
Plant Biotechnology Biological Sciences
Research subject
Biochemistry
Identifiers
urn:nbn:se:oru:diva-24347 (URN)
Conference
COST FA0804, Warszawa 5-7th September 2012
Available from: 2012-08-10 Created: 2012-08-10 Last updated: 2018-09-11Bibliographically approved
Kalbina, I., Wallin, A., Lindh, I., Engström, P., Andersson, S. & Strid, Å. (2011). A novel chimeric MOMP antigen expressed in Escherichia coli, Arabidopsis thaliana, and Daucus carota as a potential Chlamydia trachomatis vaccine candidate. Protein Expression and Purification, 80(2), 194-202
Open this publication in new window or tab >>A novel chimeric MOMP antigen expressed in Escherichia coli, Arabidopsis thaliana, and Daucus carota as a potential Chlamydia trachomatis vaccine candidate
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2011 (English)In: Protein Expression and Purification, ISSN 1046-5928, E-ISSN 1096-0279, Vol. 80, no 2, p. 194-202Article in journal (Refereed) Published
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.

Place, publisher, year, edition, pages
Academic Press, 2011
National Category
Natural Sciences Chemical Sciences
Research subject
Biochemistry
Identifiers
urn:nbn:se:oru:diva-17313 (URN)10.1016/j.pep.2011.08.010 (DOI)000296225800005 ()21903168 (PubMedID)2-s2.0-80052854898 (Scopus ID)
Projects
Ätbara vacciner
Available from: 2012-08-06 Created: 2011-09-22 Last updated: 2018-05-03Bibliographically approved
Kalbina, I., Marks, E., Lycke, N., Lindh, I., Unemo, M., Strid, Å. & Andersson, S. (2011). Construction, immunogenicity and protective efficacy in mice of a prototype chimeric Chlamydia trachomatis MOMP vaccine candidate antigen. In: : . Paper presented at 5th Biannual meeting of the Chlamydia Research Society, Redondo Beach, CA, USA, 18-21 March 2011.
Open this publication in new window or tab >>Construction, immunogenicity and protective efficacy in mice of a prototype chimeric Chlamydia trachomatis MOMP vaccine candidate antigen
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2011 (English)Conference paper, Oral presentation with published abstract (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.

National Category
Microbiology in the medical area
Research subject
Biochemistry
Identifiers
urn:nbn:se:oru:diva-24351 (URN)
Conference
5th Biannual meeting of the Chlamydia Research Society, Redondo Beach, CA, USA, 18-21 March 2011
Projects
Utveckling av slemhinneadministrerade vacciner
Available from: 2012-08-10 Created: 2012-08-10 Last updated: 2018-05-03Bibliographically approved
Kalbina, I., Wallin, A., Lindh, I., Engström, P., Andersson, S. & Strid, Å. (2011). Expression of chimeric Chlamydia trachomatis MOMP protein antigen in Arabidopsis thaliana and Daucus carota. In: Ann Depicker (Ed.), Molecular farming: plants as a production platform for high value proteins. Paper presented at COST meeting, FA0804, Gent, Belgium, 14-16 September 2011 (pp. 38-38). Bryssel: COST
Open this publication in new window or tab >>Expression of chimeric Chlamydia trachomatis MOMP protein antigen in Arabidopsis thaliana and Daucus carota
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2011 (English)In: Molecular farming: plants as a production platform for high value proteins / [ed] Ann Depicker, Bryssel: COST , 2011, p. 38-38Conference paper, Oral presentation with published abstract (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.

Place, publisher, year, edition, pages
Bryssel: COST, 2011
National Category
Plant Biotechnology
Research subject
Biochemistry
Identifiers
urn:nbn:se:oru:diva-24335 (URN)
Conference
COST meeting, FA0804, Gent, Belgium, 14-16 September 2011
Available from: 2012-08-09 Created: 2012-08-09 Last updated: 2018-05-03Bibliographically approved
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