US2004219642A1PendingUtilityA1

Methods for enhancing the production of interferon in cell culture

59
Priority: Aug 22, 1995Filed: May 28, 2004Published: Nov 4, 2004
Est. expiryAug 22, 2015(expired)· nominal 20-yr term from priority
Inventors:Allan Lau
C12N 15/67C07K 14/555C12N 9/1205C12N 2710/16643C12P 21/02C12Q 1/18C12N 2710/00088C12N 7/00
59
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Claims

Abstract

Methods for enhancing the production of interferons in animal cell culture are described. These methods rely on the manipulation of the cellular levels of certain inducers of interferon production, in particular cellular levels of double-stranded-RNA-dependent kinase (dsRNA-PKR, or PKR). In cell cultures that overproduce PKR, interferon synthesis is induced to high levels, and significant amounts of interferon can be recovered without conventional induction of interferon by virus.

Claims

exact text as granted — not AI-modified
What is claimed is:  
     
         1 . A method for producing interferon in an animal Well culture comprising: 
 (a) culturing an animal cell culture under conditions sufficient to overproduce PKR; and    (b) treating said culture to induce interferon production.    
     
     
         2 . A method for producing interferon in an animal cell culture, comprising: 
 (a) cultivating an animal cell culture under conditions sufficient to overproduce an analog of PKR; and    (c) treating said culture to induce interferon production.    
     
     
         3 . The method of  claim 2 , wherein said analog is obtainable from mouse tissue.  
     
     
         4 . The method of  claim 3 , wherein said analog is murine p65 kinase.  
     
     
         5 . The method of  claim 2 , wherein said analog is obtainable from rabbit reticulocytes.  
     
     
         6 . The method of  claim 2 , wherein said analog is obtainable from human peripheral blood mononuclear cells.  
     
     
         7 . The method of  claim 1 , wherein said PKR is human p68 kinase.  
     
     
         8 . The method of  claim 1 , wherein said interferon production is induced with a non-viral inducer.  
     
     
         9 . The method of  claim 8 , wherein said non-viral inducer is poly (I):Poly(C) or poly r(I):poly r(C).  
     
     
         10 . The method of  claim 1 , wherein said step (a) further comprises treating with a priming agent.  
     
     
         11 . The method of  claim 1 , further comprising the step of purifying the interferon produced.  
     
     
         12 . The method of  claim 1 , wherein said animal cell culture is derived from parental cells normally capable of producing interferon.  
     
     
         13 . The method of  claim 12 , wherein said overproduction of PKR is achieved by transfection of said parental cells with a vector containing DNA encoding PKR under control of a promoter.  
     
     
         14 . The method of  claim 13 , wherein said promoter is an inducible promoter.  
     
     
         15 . The method of  claim 14 , wherein said inducible promoter is a metallothionein promoter.  
     
     
         16 . The method of  claim 12 , wherein said parental cells are U937 cells.

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