Production of poxviruses with adherent or non adherent avian cell lines

Abstract

The present invention relates to a method for replicating poxviruses such as vaccinia virus comprising the steps of inoculating avian embryonic stem cells with viral particles and culturing said cells in a basal medium until cells lysis occurs and newly produced viral particles are released in said medium.

1 . A duck cell line that is infected with a virus selected in the group of orthomyxovirus, paramyxovirus and orthopoxvirus. 
     
     
         2 . The duck cell line according to  claim 1 , wherein said duck cell line replicates said virus. 
     
     
         3 . The duck cell line according to  claim 1 , wherein said virus infection comprises the steps of inoculating duck cell line with said virus and culturing said cells in a medium until cells lysis occurs and newly produced viral particles of said virus are released in said medium. 
     
     
         4 . The duck cell line according to  claim 1 , wherein said duck cell line is an embryonic derived stem cell line. 
     
     
         5 . The duck cell line according to one of  claim 1 , wherein said orthomyxovirus is influenza virus. 
     
     
         6 . The duck cell line according to  claim 1 , wherein said paramyxovirus is measles virus. 
     
     
         7 . The duck cell line according to  claim 5 , wherein said orthopoxvirus is a vaccinia virus or a recombinant vaccinia virus. 
     
     
         8 . The duck cell line according to  claim 7 , wherein the vaccinia virus is selected in the group comprising Modified Vaccinia Ankara (MVA) virus, LC16 m8, NYVAC, CVI78 or a recombinant vaccinia virus. 
     
     
         9 . The duck cell line according to  claim 7 , transfected or modified in order to produce a live vaccinia virus, a modified vaccinia virus and/or a recombinant vaccinia. 
     
     
         10 . The duck cell line according to  claim 1 , wherein said duck cell line have at least one of the following characteristics:
 a high nucleo-cytoplasmic ratio,   an endogenous alkaline phosphatase activity,   an endogenous telomerase activity,   a reactivity with specific antibodies selected from the group of antibodies SSEA-1 and EMA-1.   
     
     
         11 . The duck cell line according to  claim 1 , wherein said duck cell line have all of the following characteristics:
 a high nucleo-cytoplasmic ratio,   an endogenous alkaline phosphatase activity,   an endogenous telomerase activity, and   a reactivity with the specific antibodies SSEA-1 and EMA-1.   
     
     
         12 . The duck cell line according to  claim 1 , wherein said duck cell line are non-adherent stem cells which proliferate in suspension in a medium free of exogenous growth factors, free of serum (serum-free medium) and free of feeder cells. 
     
     
         13 . A method to produce live or attenuated, recombinant or not, vaccine comprising:
 culturing the duck cell line according to  claim 1 ;   inoculating said duck cells with viral particles;   culturing said duck cells until cells lysis occurs; and   recovering the newly produced viral particles.   
     
     
         14 . A vaccine produced by the method according to  claim 13  against acquired or infectious diseases. 
     
     
         15 . The vaccine according to  claim 14  against smallpox. 
     
     
         16 . The vaccine according to  claim 14  against influenza. 
     
     
         17 . The vaccine according to  claim 14  against measles.