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A novel chimeric MOMP antigen expressed in Escherichia coli, Arabidopsis thaliana, and Daucus carota as a potential Chlamydia trachomatis vaccine candidate
Örebro University, School of Science and Technology. (Biokemigruppen)
Uppsala universitet.
Örebro University, School of Science and Technology.
Uppsala universitet.
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2011 (English)In: Protein Expression and Purification, ISSN 1046-5928, E-ISSN 1096-0279, Vol. 80, no 2, 194-202 p.Article 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
2011. Vol. 80, no 2, 194-202 p.
National Category
Natural Sciences Chemical Sciences
Research subject
Biochemistry
Identifiers
URN: urn:nbn:se:oru:diva-17313DOI: 10.1016/j.pep.2011.08.010OAI: oai:DiVA.org:oru-17313DiVA: diva2:442794
Projects
Ätbara vacciner
Available from: 2012-08-06 Created: 2011-09-22 Last updated: 2016-11-30Bibliographically approved
In thesis
1. 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. 86 p.
Series
Örebro Studies in Life Science, 8
Keyword
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: 2011-10-18Bibliographically approved

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Kalbina, IrinaLindh, IngridAndersson, SörenStrid, Åke
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