Determination of miRNA profile related to bluetongue serotype 4-specific immune response induced by DNA vaccine in mice

Bluetongue virus is a serious hemorrhagic outlet factor of sheep, goats and other ruminants transmitted by Culicoides species mosquitoes, causing serious economic losses worldwide. Bluetongue disease is included in the category of children with mandatory warnings listed by WOAH (World Organization of Animal Health). It is the most effective measure transition to control diseases by vectorial means. Recombinant DNA technology for the immunodominant VP2 and VP5 region of this affected, Bluetongue virus, two different DNA vaccines containing VP2 or VP5, the relatedproteins of Bluetongue serotype 4 virus, and tests for miRNA profiling of the virus after immunization. For this purpose, two DNA vaccines were developed by cloning the open reading frames of VP2 and VP5 proteins into the pcDNA3.1 vector used as the DNA vaccine vector and allowing the proteins to be expressed. The developed pcDNA3.1-VP2 and pcDNA3.1-VP5 vaccine candidates are used to immunize mice. An ELISA plate specific to the relevant regions of the virus was developed and it was determined that it produced a neutralizing antibody response by indirect ELISA test. As a result of flow cytometry performed to determine cellular immunity, a significant (p<0.05) increase in IL-4 and IL-10 cytokines secreted by Th2 cells was observed in vaccinated mice compared to the control group. In the second stage of the thesis, miRNA expression profiling of bluetongue serotype 4 was performed and statistical increases and decreases in 22 miRNAs were detected using qRT-PZT.