Research About The Molecule Adjuvant Effect Of Ailuropoda Melanoleuca GM-CSF To The N Gene Of CDV DNA Vaccine

Canine distemper virus (CDV) is the unit of Paramyxovirus Virus measles virus,and caused economic losses worldwide,which become worse because of the increase of its host. Although a variety of attenuated vaccine already exists,and the Pathogenesis after Injection exists too.The nucleocapsid protein gene is the relatively conservative gene of CDV,who would cause strongly antibody response,and would be one of the chose to construct DNA vaccines, granulocyte-macrophage colony-stimulating factor (GM-CSF)is species-specific, can stimulate the body’s immune level in many ways as potential immunoadjuvant.CDV has been the threat of the giant panda during the change of pandas’living environment,but there is no vaccines of CDV to giant panda.The purpose of my experiment was to construct safe and effective DNA vaccine for giant panda.In my experiment, the nucleocapsid protein gene of CDV from eye swab of Ailuropoda melanoleuca was cloned into eukaryotic expression vector pCI-neo to construct the recombinant plasmids pCI-N and the GM-CSF gene of Ailuropoda melanoleuca was cloned for the first time,also was cloned into pCI-neo to construct the recombinant plasmids pCI-G. and to construct the expression plasmid pCI-G-N, the GM-CSF gene was cloned into the eukaryotic expression vector pCI-N through gene linker. Healthy mice were immunized with these recombinant plasmids by intramuscular injection at dosage of100μL respectively and physiological saline, pCI-neo DNA, CDV vaccine as control. Anti-CDV antibodies were measured by ELISA, lymphocyte proliferation activity was detected by the amount of the IL-4and IFN-y. The results showed that and the DNA vaccine caused high level immune response,and the contents of IL-4,IFN-yand anti-CDV antibodies of the mice in pCI-N+pCI-G group at56days after the first injection was82.54pg/ml、317.32pg/m、249.57pg/ml respectively. The result indicated that the recombination protein can express normally,the CDV vaccine constructed in the experiment can stimulate mice to produce the CDV-specific antibody, and the GM-CSF can enhance the immunity of CDV DNA vaccine as cytokine adjuvant. The results provide reference data for the new generation vaccine research of CDV.