Vaccine Against Disease
Grafting of B- and T-cell Epitopes of Foot-and-Mouth Disease Virus to Recombinant Protein Caf1 of Yersinia Pestis and Use of its Construction as a Vaccine Against Foot-and-Mouth Disease
Tech Area / Field
- AGR-VTH/Vaccines and Theraupetics/Agriculture
3 Approved without Funding
Institute of Immunological Engineering, Russia, Moscow reg., Lyubuchany
Project summaryFood-and-mouth disease virus (FMDV) causes a highly contagious, economically important disease of cloven-hoofed animals that affects countries with the highly developed cattle breeding. The prevention of FMD outbreaks is based on quarantine measures and immunization of livestock with killed virus vaccines. Present-day FMD vaccines are effective for the disease control but they are based on the technology of large-scale cultivation of the highly infectious virus. A high percentage of FMD outbreaks in Europe have been caused by insufficiently inactivated vaccine virus or by infectious virus escape from vaccine production plants. Since 1981 a new policy of non-use of traditional vaccines and prohibition of works with infectious virus in a number of countries has contributed to the development of synthetic vaccines against FMD. These vaccines are based on the peptide synthesis, or recombinant DNA technologies. At present the production of cheaper, safer and more stable vaccines of a new generation is a matter of great urgency.
Within the framework of the project we are going to use artificially synthesized genetic structures of FMDV encoding immunogenic viral B- and T-cell epitopes responsible for protective immunity formation. To express these genetic structures it is expected to construct both vectors for intracellular biosynthesis and vectors using secretory mechanisms of bacterial cells. In this connection the use of secretory signals of the capsular antigen Cafl of Yersinia pestis is of great interest. The capsular antigen Cafl has pronounced immunogenic properties and can be an effective carrier of FMDV antigenic determinants. Using protein-engineering methods we intend insert one of the imunnogenic B- (T)-cell epitopes of FMDV instead of the hydrophilic loop detecting the B-cell epitope on the surface of the Cafl protein subunit. We have cloned and sequenced the capsular Yersinia pestis fl operon which consists of (i) the cafl gene of the capsular protein Cafl, (li) the caflm gene of the molecular periplasmic chaperone CaflM, (in) the cafl a gene of the usher-protein Cafl A, and (iv) the caflr gene controlling the process for the capsule formation. The fl operon of Yersinia pestis will supposedly provide the superproduction and secretion of Cafl with a grafted construction in Eschehchia coli cells. This recombinant vaccine may be an efficient tool for the treatment and prevention of the dangerous disease of farm animals caused by FMDV.
The project is based on the knowledge of (i) B- and T-cell epitopes of Wl FMDV responsible for the immunoprotection formation; (ii) structural-functional properties of the capsular antigen Cafl of Yersinia pestis; (iii) functions of specific molecular periplasmic chaperones of pathogenic gram-negative bacteria.
Development of new approaches to the construction of recombinant vaccine against FMD based on the capsular protein Cafl of Yersinia pestis and immunogenic B- and (T)-cell epitopes of FMDV.
New recombinant protein constructions Cafl containing viral B- and T-cell epitopes would be obtained. These peptide constructions would be safe and effective for animal protection against FMD. New approaches to the development of the recombinant vaccine using genetic elements of the fl operon of Yersinia peslis would be studied. The recombinant producing strain of the virus-specific protein would be designed. This strain would form the basis for a new vaccine against FMD.
Technical Approach and Methodology
To perform the tasks defined in this project, it is planned to use a wide set of currently available research methods, including theoretical conformation analysis, genetic and protein engineering methods, CD- and UV-spectroscopy, electrophoresis in PAGE, enzymimmunoassay techniques, cell- and humoral immunity methods and some other physico-chemical, immunochemical and biochemical methods.
The general strategy would be the creation of active genetically engineered constructions containing the gene of capsular protein Cafl with different fragments of the main immunogen VPl of FMDV. Immunogenic and protective properties of recombinant protein would be studied in laboratory and sensitive animals.
International co-operation under the project submitted to the ISTC.
In order to establish new contacts and for further integration into the international scientific collaboration network, the following activities would be scheduled within the framework of this project:
- elaboration of common research strategy and collaborative programmes;
- exchange of information and materials required for successful research,
- formation of personal in specific areas of partner competence, mobility programme;
- organisation of meetings and seminars with participation of collaborators;
- elaboration of assistance programme for Russian partner (consultations, advice, preparation of publications, provision of computer modeling programs of molecular structures);
- preparation of new research projects and research of potential foreign partnership via corresponding collaborator.
Potential partnership would be established with Dr A.I. Donaldson (Institute for Animal Health, Pirbright Laboratory, UK) who has been engaged in research of specific prevention of FMD for a long time and would take an interest in collaborative investigations within the framework of the project.