Study On Immune Effects Of Porcine Transmissible Gastroenteritis Virus With Nano Silicon Adjuvant

Porcine Transmissible Gastroenteritis(TGE)is one of high exposure to infectious diseases caused by Transmissible Gastroenteritis Virus(TGEV)in porcine,the significant clinical symptoms of this disease are severe diarrhea,vomiting,dehydration,etc.Pigs of different ages are all susceptible,and to newborn pialets,it is highly lethal,especially to the ages of piglets within two weeks,it can be up to 100%.There are no effective treatment measures after its attacked by TGEV,resulting in decreased feed efficiency and production degradation,and it caused a huge economic loss to the pig farming industry.Therefore,it is proved that scientific Immunity by the vaccine is the most effective measure to TGE in clinical prevention and control.In this experiment,the immune effect on mice after immunization by TGEV inactivated vaccine with different adjuvants are mainly studied,and the expression of miRNA related to immune is explored preliminarily,providing greater possibilities for the selection of high-quality vaccine adjuvants,and providing certain experiment basis for clinical prevention and disease prevention.The details of this research are as follows:1.First,the strain of TGEV HN-2012 is inactivated by 0.2%formaldehyde,and preparing TGEV oil emulsion inactivated vaccine by proportion of 1:2 between inactivated venom and oil emulsion,then according to the same proportion,preparing nano silicon dioxide adjuvants inactivated vaccines with the particle size of 70 nm and 150 nm.Upon examination,both physical and chemical properties and aseptic of prepared three kinds of inactivated vaccine are qualified,and the safety of rats is good,too.2.Using three kinds of prepared inactivated vaccine to immune mice,and then comparing its immune effect with the TGEV without adjuvant inactivated vaccine and commercially available TGEV inactivated vaccine with aluminum hydroxide adjuvant respectively.After immune 0,1,2,3,4,5,6,8 and 10 weeks,the immune level of humoral immunity and cellular immunity in mice through ELISA and flow cytometry respectively.Lymphocyte proliferation is detected by MTT colorimetry,and the results show:all 5 kinds of vaccine can induce mice to generate specific humoral immunity and cellular immune response.Furthermore,Group 70 nm、Group 150 nm and Group TGEV oil emulsion can induce to produce a higher level of humoral immunity in mice;Group 70 nm can induce to produce a higher level of cellular immunity in mice.In addition to B lymphocytes of 150 nm group between saline group are different significantly,and the remaining difference among lymphocyte proliferation condition is not significant.3.To different adjuvants inactivated vaccine groups,expression of miR-21,miR-27,miR-30,miR-146,miR-150,miR-155 and miR-181 related to mice immune are discussed preliminarily.After immune 0 h,12 h and 72 h,relative expression index of miRNA related to mice immune is detected using fluorescence quantitative PCR method.The results show that after 12 h,72 h,the relative expression of immune related genes decrease,but decreasing condition is not consistent.And all relative expression of miR-181 is low.At 12 h,the expression level of miR-27,miR-146 and miR-155 of Group 70 nm are higher than other groups;and the expression of miR-30,miR-150 and miR-181 of Group 150 nm are higher than other test groups.At 72 h relative expression of all miRNA of Group TGEV oil emulsion are more higher than other inactivated vaccine groups with different adjuvants.