Imaging methods and compositions comprising fluorescent dyes associated with viral components for nerve imaging

Abstract

Disclosed herein are compositions and methods for imaging nerve cells. The composition comprises a fluorescent dye; and a viral component selected from a neurotropic, replication-defective virus, a viral protein of a neurotropic virus, and a capsid of a neurotropic virus. Although the fluorescent dye in itself cannot penetrate nerve cells, the fluorescent dye is bound to the viral component to form a dye/viral component complex that is capable of penetrating nerve cells.

1 .- 11 . (canceled) 
     
     
         12 . A method of making a composition, comprising:
 combining a fluorescent dye with a viral component selected from a neurotropic, replication-defective virus, a viral protein of a neurotropic virus, and a capsid of a neurotropic virus,   allowing the fluorescent dye to be bound to the viral component to form a dye/viral component complex that is capable of penetrating nerve cells.   
     
     
         13 . The method of  claim 12 , wherein the viral component is a neurotropic, replication-defective virus. 
     
     
         14 . The method of  claim 13 , wherein the virus is rendered replication-defective by mutation, UV radiation, chemical treatment, or heat treatment. 
     
     
         15 . A method for reducing the risk of iatrogenic injury to a subject during a surgical procedure, comprising:
 (a) administering a composition to the subject, the composition comprising:
 a fluorescent dye; and 
 a viral component selected from a neurotropic, replication-defective virus, a viral protein of a neurotropic virus, and a capsid of a neurotropic virus, 
 wherein the fluorescent dye is bound to the viral component to form a dye/viral component complex; 
   (b) allowing the dye/viral component complex to penetrate nerve cells;   (c) applying a sufficient amount of radiant energy to the subject such that the dye fluoresces;   (d) intraoperatively obtaining a fluorescence image of the subject; and   (e) observing the fluorescence image to view one or more nerves in the subject.   
     
     
         16 . The method of  claim 15 , wherein the fluorescent dye is selected from indocyanine green, infracyanine green, green fluorescent dye, fluorescein isothiocyanate, rhodamine, phycoerythrin, phycocyanin, allophycocyanin, o-phthaldehyde, fluorescamine, Rose Bengal, trypan blue, fluoro-gold, 3-indocyanine-green-acyl-1,3-thiazolidine-thione, green fluorescent protein, red fluorescent protein, yellow fluorescent protein, blue fluorescent protein and other fluorescent proteins. 
     
     
         17 . The method of  claim 16 , wherein the fluorescent dye is indocyanine green. 
     
     
         18 . The method of  claim 15 , wherein the composition comprises replication-defective herpesvirus. 
     
     
         19 . The method of  claim 18 , wherein the herpesvirus is HSV-2. 
     
     
         20 . The method of  claim 18 , wherein the replication-defective virus comprises HSV-2 ICP10ΔRR. 
     
     
         21 . A method for diagnosing a nerve condition, comprising:
 (a) administering a composition to the subject, the composition comprising:
 a fluorescent dye; and 
 a viral component selected from a neurotropic, replication-defective virus, a viral protein of a neurotropic virus, and a capsid of a neurotropic virus, 
 wherein the fluorescent dye is bound to the viral component to form a dye/viral component complex; 
   (b) allowing the dye/viral component complex to penetrate nerve cells;   (c) applying a sufficient amount of radiant energy to the subject such that the dye fluoresces;   (d) intraoperatively obtaining a fluorescence image of the subject; and   (e) observing the fluorescence image to determine whether the nerve is transected.