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Feeding of mice with Arabidopsis thaliana expressing the HIV-1 subtype C p24 antigen gives rise to systemic immune responses
Örebro University, School of Science and Technology.
Örebro University, School of Science and Technology.
Örebro University, School of Science and Technology.ORCID iD: 0000-0001-9713-2365
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2008 (English)In: Acta Pathologica, Microbiologica et Immunologica Scandinavica (APMIS), ISSN 0903-4641, E-ISSN 1600-0463, Vol. 116, no 11, p. 985-994Article in journal (Refereed) Published
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.

Place, publisher, year, edition, pages
Oxford: Blackwell , 2008. Vol. 116, no 11, p. 985-994
National Category
Medical and Health Sciences Immunology in the medical area
Research subject
Immunology
Identifiers
URN: urn:nbn:se:oru:diva-4631DOI: 10.1111/j.1600-0463.2008.00900.xOAI: oai:DiVA.org:oru-4631DiVA, id: diva2:138930
Available from: 2008-10-14 Created: 2008-10-14 Last updated: 2018-01-13Bibliographically approved
In thesis
1. Development of transgenic Arabidopsis thaliana and Daucus carota plant producing the p24 capsid protein from HIV-1 subtype C for the use as an edible plant-based vaccine model
Open this publication in new window or tab >>Development of transgenic Arabidopsis thaliana and Daucus carota plant producing the p24 capsid protein from HIV-1 subtype C for the use as an edible plant-based vaccine model
2009 (English)Licentiate thesis, comprehensive summary (Other academic)
Place, publisher, year, edition, pages
Örebro: , 2009. p. 56
Series
Licentiate theses in Life Science ; 7
National Category
Biochemistry and Molecular Biology Biochemistry and Molecular Biology Natural Sciences Chemical Sciences
Research subject
Biochemistry
Identifiers
urn:nbn:se:oru:diva-8700 (URN)
Presentation
2009-04-29, Hörsal M, Örebro universitet, Örebro, 00:00 (English)
Opponent
Supervisors
Projects
Production of edible vaccines in plants
Available from: 2010-02-01 Created: 2009-12-08 Last updated: 2017-10-18Bibliographically approved
2. Plant-produced STI vaccine antigens with special emphasis on HIV-1 p24
Open this publication in new window or tab >>Plant-produced STI vaccine antigens with special emphasis on HIV-1 p24
2011 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

Objective: To establish stable transgenic non-toxic plants as a platform for plant-based vaccine production as well as potential oral delivery system of vaccine antigens for sexually transmitted infections (STIs). The concept is to immunize the mucosal immune system present in the gut-associated lymphoid tissues (GALT). HIV-1 p24 subtype C protein has been used as the main antigen model, in parallel with an engineered unique chimeric MOMP antigen from Chlamydia trachomatis serovar E.

Methods: Chimeric MOMP and p24 vaccine antigens were successfully inserted into the nuclear genomes of Arabidopsis thaliana and Daucus carota via Agrobacterium-mediated gene transfer. The characteristics of the genetic inserts and corresponding mRNAs and recombinant proteins in planta were described using several methods, including northern, Southern, and western blotting, ELISA, and a commercial HIV Ag/Ab combination assay. Immunogenicity of the antigens was studied in mice models.

Results: Transgenes of both plant species expressing p24 or chimeric MOMP were successfully generated. Additional HIV-1 vaccine antigen candidates were introduced and the genetic inserts have been confirmed in Arabidopsis thaliana. The Arabidopsis thaliana expressing p24 and chimeric MOMP were demonstrated to be stable over generations and antigenicity analyses showed that plant-derived HIV-1 p24 and chimeric MOMP retained immunological epitopes when they were expressed in planta. Oral administration of transgenic plant material generated a priming effect of the immune competent cells present in the GALT, shown by the presence of antigen-specific-IgG in mice sera after boosting. Mice immunized with plant-derived HIV-1 p24 antigen were also analyzed for antigen-specific faecal IgA as well as cellular immune responses. However, detectable levels of the two latter immune responses were not observed. The Chlamydia trachomatis chimeric MOMP antigen was further evaluated for its potential as a vaccine antigen candidate, with positive results indicating a more rapid clearance of the Chlamydia trachomatis infection post immunization.

Conclusion: Stable non-toxic transgenic plants expressing either HIV-1 p24 or a novel  Chlamydia trachomatis chimeric MOMP antigens have successfully been developed. The two plant-produced STI vaccine antigens have in initial mice feeding studies provided important proof-of-concept for the oral vaccination approach. Now, immunization studies to expand, en-hance, and improve knowledge of the immune responses generated by the orally delivered transgenic plants are of high priority.

Place, publisher, year, edition, pages
Örebro: Örebro universitet, 2011. p. 86
Series
Örebro Studies in Life Science ; 8
Keywords
Plant-based vaccines, Arabidopsis thaliana, HIV-1, p24, GALT, mucosal immunity, Chlamydia trachomatis, MOMP, Daucus carota
National Category
Chemical Sciences
Research subject
Chemistry
Identifiers
urn:nbn:se:oru:diva-17242 (URN)978-91-7668-817-5 (ISBN)
Public defence
2011-10-28, Hörsal M, Örebro universitet, Fakultetsgatan 1, Örebro, 10:15 (English)
Opponent
Supervisors
Projects
Kemi/biokemi
Available from: 2011-09-13 Created: 2011-09-13 Last updated: 2017-10-17Bibliographically approved

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Lindh, IngridKalbina, IrinaScherbak, NikolaiSävenstrand, HelenaStrid, ÅkeAndersson, Sören

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