Genetically engineered cell lines and systems for propagating varicella zoster virus and methods of use thereof

Abstract

The present invention provides genetically engineered cell lines, recombinant vectors, and vaccines. The present invention also provides methods for generating an in vitro system for Varicella zoster virus (VZV), and the in vitro systems generated by these methods. The present invention further provides methods for reactivating VZV, and VZV reactivated by these methods. Finally, the present invention provides a method of screening for an agent for treating VZV infection.

1 . A vaccine comprising an attenuated live virus produced by culturing genetically-engineered cells stably transformed with a nucleotide sequence encoding at least two full-length mannose-6-phosphate receptors and a pharmaceutically acceptable carrier. 
     
     
         2 . A method for generating an in vitro system for  Varicella zoster  virus (VZV), comprising the steps of:
 (a) isolating enteric ganglia from guinea pig; and   (b) contacting the enteric ganglia with cell-free VZV to generate latent expression of the VZV.   
     
     
         3 . An in vitro system for VZV generated by the method of  claim 2 . 
     
     
         4 . The in vitro system of  claim 3 , wherein the method further comprises the step of reactivating the VZV by contacting the system with a vector containing a nucleic acid sequence coding for VZV ORF61 or a homologue thereof. 
     
     
         5 . The in vitro system of  claim 4 , wherein the homologue is HSV ICP0. 
     
     
         6 . An in vitro system for  Varicella zoster  virus (VZV), comprising an enteric ganglion that has been contacted with cell-free VZV to produce a latent VZV, wherein the VZV is subsequently reactivated to express VZV in an active form. 
     
     
         7 . A method for reactivating  Varicella zoster  virus (VZV), comprising the steps of:
 (a) isolating enteric ganglia from guinea pig;   (b) contacting the enteric ganglia with cell-free VZV to generate latent expression of the VZV; and   (c) contacting the infected ganglia with a vector containing a nucleic acid sequence encoding VZV ORF61 or a homologue thereof.   
     
     
         8 . The method of  claim 7 , wherein the homologue is HSV ICP0. 
     
     
         9 . A  Varicella zoster  virus reactivated by the method of  claim 10 . 
     
     
         10 . A method of screening for an agent for treating  Varicella zoster  virus (VZV) infection, comprising use of an in vitro system for VZV, wherein the in vitro system comprises an enteric ganglion that has been contacted with cell-free VZV to produce a latent VZV, and wherein the VZV is subsequently reactivated to express VZV in an active form.