Methods for attenuating virus strains for diagnostic and therapeutic uses

Abstract

Lister strain vaccinia viruses that are encode a modified A34R gene resulting in increased extracellular enveloped virus (EEV) form of the virus and/or increased resistance of the virus to neutralizing antibodies. The viruses can be used in methods of treatment of diseases, such as proliferative and inflammatory disorders, including as anti-tumor agents.

What is claimed: 
     
         1 . A recombinant Lister strain vaccinia virus that comprises replacement of the A34R gene with the A34R gene from another vaccinia virus strain. 
     
     
         2 . The recombinant vaccinia virus of  claim 1 , wherein the A34R gene is replaced by the A34R gene from vaccinia IHD-J strain. 
     
     
         3 . The recombinant vaccinia virus of  claim 1 , wherein the replacement increases the extracellular enveloped virus (EEV) form of vaccinia virus and/or increases the resistance of the virus to virus neutralizing antibodies. 
     
     
         4 . The Lister strain vaccinia virus of  claim 1  that comprises deletion of the A35R gene. 
     
     
         5 . The recombinant virus of  claim 1  that is an LIVP strain. 
     
     
         6 . The recombinant virus of  claim 5  that is a derivative of the virus designated GLV-1h68. 
     
     
         7 . The recombinant virus of  claim 6  that is the virus designated GLV-1i69. 
     
     
         8 . A recombinant virus of  claim 1 , further comprising one or more heterologous nucleic acid molecules that encode(s) a diagnostic and/or therapeutic protein. 
     
     
         9 . The recombinant virus of  claim 1 , comprising a nucleic acid molecule that encodes a diagnostic protein that is a detectable protein or a protein that induces a detectable signal. 
     
     
         10 . The recombinant virus of  claim 9 , wherein the diagnostic protein is selected from among a luciferase, a fluorescent protein, an iron storage molecule, an iron transporter, an iron receptor or a protein that binds a contrasting agent, chromophore or a compound or detectable ligand that can be detected. 
     
     
         11 . The recombinant virus of  claim 8 , that encodes a therapeutic protein selected from among a cytokine, a chemokine, an immunomodulatory molecule, an antigen, a single chain antibody, antisense RNA, prodrug converting enzyme, siRNA, angiogenesis inhibitor, a toxin, an antitumor oligopeptide, a mitosis inhibitor protein, an antimitotic oligopeptide, an anti-cancer polypeptide antibiotic, and a tissue factor. 
     
     
         12 . The virus of  claim 2 , that is modified, wherein the unmodified virus is selected from among GLV-1h22, GLV-1h68, GLV-1h70, GLV-1h71, GLV-1h72, GLV-1h73, GLV-1h75, GLV-1h81, GLV-1h82, GLV-1h83, GLV-1h84, GLV-1h85, GLV-1h86, GLV-1j87, GLV-1j88, GLV-1j89, GLV-1h90, GLV-1h91, GLV-1h92, GLV-1h96, GLV-1h97, GLV-1h98, GLV-1h104, GLV-1h105, GLV-1h106, GLV-1h107, GLV-1h108 and GLV-1h109. 
     
     
         13 . A pharmaceutical composition comprising a recombinant virus of  claim 1  in a pharmaceutically acceptable carrier. 
     
     
         14 . The pharmaceutical composition of  claim 13  that is formulated for local or systemic administration. 
     
     
         15 . A method of treatment, comprising administering the pharmaceutical composition of  claim 13  to a subject to effect the treatment of a tumor, cancer or metastasis. 
     
     
         16 . A pharmaceutical composition comprising a recombinant virus of  claim 12  in a pharmaceutically acceptable carrier. 
     
     
         17 . A method of treatment, comprising administering the pharmaceutical composition of  claim 16  to a subject to effect the treatment of a tumor, cancer or metastasis.