Influence Of Reticuloendotheliosis Virus To Apoptosis And P53 Expression In Chick Embryo Fibroblast

Reticuloendotheliosis(RE)caused by Reticuloendotheliosis virus(REV)is a kind of immunosuppressive and tumor disease which can mainly cause immunosuppressive,reticuloendothelial cell hyperplasia,lymphomagenesis in infected chicks.Apoptosis is a kind of programmed death process and plays an important role in maintaining stabilization in internal environment.P53 is not only closely relevant with the occurence and development of tumor,but also is closely related to the occurence of apoptosis.Now,the epidemiology,nosetiology,specific diagnosis,especially in the molecular biological characteristics of REV are researched systemicly,and some results are achieved,but the understanding about immunosuppressive symptom,especially in the mechanism of immunosuppressive symptom are lacked.To state the mechanism of immunosuppressive symptom,and disscuss the relation of apoptosis and pathopoiesia of REV.In this experiment,SPF chick embryo fibroblast(CEF)was choosed as research subject,and divide into control group(C group)and REV infected group(I group),thereinto,CEF of REV infected group was infected 2h by 100TCID50 concentration REV,adding DMEM with 2% fetal bovine serum after abandoning infecting liquid,but CEF of control group was acted 2h without REV,adding DMEM with 2% fetal bovine serum after abandoning DMEM,on the basis of preparing rabbit polyclonal antibody against chicks P53,applying the experiment methods of flow cytometry,Real time-PCR and Western Blot,respectively,according to detect the apoptosis number,the expression of p53 mRNA and the changing of protein content of CEF of I group and C group,the dynamic changing of the value above mentioned in CEF was researched roundly and systematacially in different detection time after infected with REV.Supply scientific experimental theoretical basis to elaborate the mechanism of apoptosis caused by REV and reference frame to elaborate immunosuppressive symptom induced by REV from the point of view of apoptosis.The experimental results were as following:1.Rabbit polyclonal antibody against chicks P53 is prepared successfully,the ELISA titer of this antibody is 219,the specificity detected by Western Blot is high.2.Within 72 h after chick embryo fibroblast infected with REV,the apoptosis number of SPF chick embryo fibroblast is higher than uninfected with REV control group,especially infected with REV group is remarkable higher than control group CEF in 72 h after infection effect(P<0.05).3.In 6h-72 h after chick embryo fibroblast infected with REV,compared to uninfected with REV control group,the expression of p53 m RNA of infected group is higher in different degree,especially the expression of p53 mRNA of infected group is remarkable higher than control group in 48 h and 72 h after infection effect(P<0.05).4.In 6h-72 h after chick embryo fibroblast infected with REV,the content of P53 protein in infected group is higher than control group in different degree.Draw conclusion according to exprement result: REV can increase the number of apoptosis of SPF CEF,the expression of p53 m RNA and P53 protein,it indicates that the increasing of P53 protein contein and the increasing of apoptosis numbers induced REV are positive correlation.