Methods for enhanced virus-mediated DNA transfer using molecules with virus-and cell-binding domains

Abstract

A method to increase the efficiency of transduction of hematopoietic and other cells by retroviruses includes infecting the cells in the presence of fibronectin or fibronectin fragments. The fibronectin and fibronectin fragments significantly enhance retroviral-mediated gene transfer into the cells, particularly hematopoietic cells including committed progenitors and primitive hematopoietic stem cells. The invention also provides improved methods for somatic gene therapy capitalizing on enhanced gene transfer, hematopoietic cellular populations, and novel constructs for enhancing retroviral-mediated DNA transfer into cells and their use.

1 - 2 . Canceled  
     
     
         3 . A viable cellular population produced by a method comprising infecting the cells with a retrovirus in the presence of an effective immobilized amount of material including a ligand which binds to the cells and a ligand which binds to the retrovirus, so as to co-localize the retrovirus and the cells and increase the transduction efficiency of the cells, said infecting being conducted in a medium essentially free from hexadimethrine bromide.  
     
     
         4 . The viable cellular population of  claim 3  which comprises hematopoietic stem cells.  
     
     
         5 . A method for cellular grafting, comprising: grafting a mammal with a viable cellular population produced by a method comprising infecting the cells with a retrovirus in the presence of an effective immobilized amount of material including a ligand which binds to the cells and a ligand which binds to the retrovirus, so as to co-localize the retrovirus and the cells and increase the transduction efficiency of the cells, said infecting being conducted in a medium essentially free from hexadimethrine bromide.  
     
     
         6 - 7 . Cancelled  
     
     
         8 . A method for cellular grafting, comprising: grafting a mammal with a cellular composition, comprising a substantially retroviral-transduced population of viable cells, said composition being essentially free from both retroviral producer cells and hexadimethrine bromide.  
     
     
         9 . The method of  claim 8  wherein the cellular population comprises hematopoietic stem cells.  
     
     
         10 - 15 . Cancelled  
     
     
         16 . The method of  claim 3 , wherein the medium is essentially free from any polycationic agent which increases the efficiency of transduction of the viable mammalian cells by the retrovirus in co-culture, but which agent reduces the efficiency of transduction of the cells by the retrovirus in the presence of said material.  
     
     
         17 . The method of  claim 16 , wherein said material comprises substantially pure fibronectin, substantially pure fibronectin fragments, or mixture thereof.  
     
     
         18 . The method of  claim 5 , wherein the medium is essentially free from any polycationic agent which increases the efficiency of transduction of the viable mammalian cells by the retrovirus in co-culture, but which agent reduces the efficiency of transduction of the cells by the retrovirus in the presence of said material.  
     
     
         19 . The method of  claim 18 , wherein said material comprises substantially pure fibronectin, substantially pure fibronectin fragments, or mixture thereof.  
     
     
         20 . The method of  claim 8 , wherein said composition is essentially free from any polycationic agent that is effective to increase the efficiency of transduction of the viable cells by the retrovirus in co-culture.  
     
     
         21 . The method of  claim 20 , wherein said viable cells have been transduced in the presence of substantially pure fibronectin, substantially pure fibronectin fragments, or mixture thereof, so as to increase the efficiency of transduction by the retrovirus.