Anti-Hantavirus DNA Vaccine
Immunogenicity of the Experimental Aerosol Anti-Hantavirus DNA Vaccine Preparation
Tech Area / Field
- BIO-CGM/Cytology, Genetics and Molecular Biology/Biotechnology
3 Approved without Funding
Research Center of Toxicology and Hygienic Regulation of Biopreparations, Russia, Moscow reg., Serpukhov
- Chumakov Institute of Poliomyelitis and Viral Encephalitides of Russian Academy of Medical Sciences, Russia, Moscow
Project summaryThe project objective is to solve 2 tasks: develop the accurate method for DNA-vaccine dosage estimation and molecular-immunological analysis of cellular component of immune response, which could contribute considerably to general response to DNA vaccine application. The mentioned tasks arose during the research conducted on the ISTC #1813ð Project funded by DTRA, which resulted in experimentally justified conclusion on the potential of inhalation delivery of DNA-vaccine preparation against Hantavirus infection for induction of immune response in laboratory animals at the natural entrance of the infection, which provides a basis for quick designing of anti-Hantavirus DNA-vaccine, the alternative to which, a classical protein vaccine, is not available yet. Thus, before the DNA vaccine is offered and becomes the object of pre-clinical trials, the two above mentioned problems should be solved. These main tasks are planned to be fulfilled in the framework of the proposed project.
To solve the first problem, development of the method for efficient dissociation of PEI/DNA complex will be needed. The second one needs development of the model for evaluating the efficiency of cellular response to DNA/PEI preparation.
For the development of the model by method of retro-virus transfection, lines of immortalized murine fibroblasts C57SV(H-2b) and D2SV(H-2d) expressing a minigen coding a fragment of virus glycoprotein G1 of Puumala AA1-259 strain containing H-2b- and H-2d-restricted virus epitopes will be obtained. The retro-virus transfection of the indicated fibroblasts will be performed using retro-virus vector pMSCVMigR1(IRES-GFP). Cloning of GFP+ cells will be conducted. The produced genes will be tested for expression of virus CTL-epitopes by survival of C57SV(H-2b) and D2SV(H-2d) mice after transplantation of transfected fibroblasts of cell lines H-2b and H-2d, respectively. Survived mice will be tested for the presence of virus-specific memory CD8+ cells by proliferation in a mixed culture of lymphocytes and tumor cells MLTC with transfected or intact control tumor cells subjected to severe temperature shock.
The obtained lines expressing virus epitopes will be used as a test antigen for induction of specific antivirus immunity (positive control).
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