| Equid herpesvirus 1(EHV-1)is one of the main pathogens causing respiratory diseases in horses.It can also cause equine neurological diseases,miscarriage of pregnant horses and death of newborn foals.The diseases caused by it are prevalent all over the world and cause serious economic losses to the horse industry.The epidemic situation of EHV-1 in China is severe and there is no specific drug treatment,and the prevention and control of EHV-1 mainly relies on vaccine immunization.At present,there is no commercial vaccine in China,so it is of great significance to develop a new vaccine to prevent and control the epidemic of EHV-1.1.The EHV-1 XJ2015 gp2 gene deletion strain has been constructed in our laboratory in the early stage.By proliferating in RK-13 cells,the size of plaque was observed,and the high quality strain was selected to calculate the half infection amount of the cells.The virus was concentrated by saturated ammonium sulfate precipitation method,then centrifuged at ultra-high speed,and further purified by sucrose density gradient centrifugation method.The results of electron microscopy showed that the virus particles remained intact after purification,and the diameter of the virus particles was 100～120 nm.The highest efficiency of the virus particles after concentration could reach 1×108.72 TCID50/0.1 mL.The concentration of 0.02%beta-c lactone inactivated virus 24 hours A day,add A adjuvant mixed with inactivated viruses,the preliminary preparation of inactivated vaccines,configuration,respectively,106 TCID50.1mL,107 TCID50/0.1 mL,108 TCID50/0.1 mL different virus titer of inactivated vaccine,and to quality inspection of inactivated vaccines,including vaccines,sterility testing physical properties test,safety test.The results showed that the bacteria,fungi and molds of the vaccine were all negative.Under different conditions,the physical morphology is maintained in normal state,and all physical tests are up to the standard.By vaccinating Syrian mice of different ages,no adverse reactions were observed in the vaccinated mice within 14 days.2.Through to the Syrian muscle in mice inoculated EHV-1 XJ2015 Δ gp2 inactivated vaccine strains,evaluation Δ gp2 inactivated vaccines each between experimental group and EHV-1 XJ2015 inactivated vaccine strains of antibody level differences,and to set up the PBS control group.The results showed that there was no significant difference between the vaccine groups after the first immunization.After the second immunization,the antibody level of the experimental groups showed an upward trend with the increase of immunization time and times.The third time after immune EHV-1 vaccine group and Δ gp2 antibody level is extremely significant difference between groups.The results showed that the body weight of the vaccine immunization group did not fluctuate significantly 10 days after the challenge,and no obvious clinical symptoms were shown in the vaccine immunization group.In contrast,the control group showed a decreasing trend in body weight,developed symptoms such as dyspnea and nervous tremor,and died on the 9th day.Through autopsy to pathology observation,each test in mice,according to the results of Δ gp2 vaccine group and EHV-1 mice lungs and brain tissue vaccine group were not found obvious pathological changes,pathological observation not present histologic changes;While in the control group,there was brain edema,lung swelling and bleeding.Pathological sections could observe the damage of alveolar structure,obvious thickening of alveolar wall,and accumulation of inflammatory granulocytes and macrophages in the brain tissue..In summary,Δ gp2 inactivated vaccine was prepared using EHV-1 XJ2015 Δgp2 gene deletion strain in this study,and its immune effect was evaluated using the Syrian mouse model.The results showed that the vaccine provided a good protective effect,and the mice produced a good antibody level after immunization.This study provides data support for the later development of novel EHV-1 inactivated vaccine. |
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