Study On The Biological Characteristics And Preservation Stability Of Recombinant Oncolytic Herpes Simplex Virus Type â…¡ (OH2)

Objective:Study recombinant oncolytic herpes simplex virus type II(OH2) on the preservation stability and biological characteristics stability containing the growth curves,the hGM-CSF experssed,the oncolytic activity in vitro and vivo,to provide theoretical basis for clinical application.Methods:We cultured OH2 in vero cells for 20 generations to detect biological characteristics of OH2 4th,10 th,20th. Virus growth curves were revealed through a series of methods including cell culturing,virus culturing and titer determination.The contents and activity of hGM-CSF expressed by OH2 were detected by using enzyme linked immunosorbent assay(ELISA) and human erythrocyte leukemia lymphocyte(TF-1)culturing methods.Further experiments such as cell proliferation toxicity test and tumor inhibition test were did to prove the antitumor activity of OH2 4th,10 th,20t hin vivo and in vitro.The purified OH2 kept in stabilizer was stored under different temperatures and the titers of OH2 stored were determined at different times.Results:There are no obvious difference among OH2 4th,10 th,and 20 th on growth curves,the cancer cell killing ability in vitro,the content and activity of hGM-CSF expressed.Moreover,there are significant anti-tumor effects for OH2 of different generations including 4th,10 th,20th,the tumor inhibition rates of which are90.4%,74.1%,79.9% respectively.The titer of virus preserved in stablizer is relatively stable.After preserved in4℃,-20℃,-80℃ for 25 months,titers of OH2 in high titer group remained stable basically,however titers in low titer group declined for 100 times.Titers of OH2 of high titer group and low titer group kept in 20℃ reduced respectively from 107.98 CCID50/ml and107.05 CCID50/ml to 107.5 CCID50/ml and 105 CCID50/ml afte stored for 100 days.Conclusions:(1) After passaged for 20 generations continuously,the biological characteristics of OH2 remained stable.(2) Stablizer had a good preservation effect for OH2.The viability of virus of high titer could maintain more durable than virus of low titer when kept in stablizer.Temperatures such as 4℃,-20℃,-80℃ were more suitable...