Methods for enhanced virus-mediated DNA transfer using molecules with virus-and cell-binding domains
AssigneeAdvanced Research and Technology Institute, Inc.
InventorDavid A. Williams
Life of this patent
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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.
Claims19
A method for obtaining a transduced population of viable cells by a retrovirus, comprising:
- infecting the cells with a retrovirus in the presence of an effective immobilized amount of fibronectin, a fibronectin fragment, or a mixture thereof to increase the efficiency of transduction of the cells by the retrovirus, said fibronectin, fibronectin fragment, or mixture thereof 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, said infecting being conducted in a medium essentially free from a polycationic agent which increases the efficiency of transduction of the 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 fibronectin, fibronectin fragment, or mixture thereof.
A cellular composition, comprising:
- a substantially retroviral-tranduced in vitro population of viable cells, at least about 20% of which are retroviral-transduced, said composition being essentially free from both retroviral producer cells and a polycationic agent which increases the efficiency of transduction of the cells by the retrovirus in co-culture, said composition further comprising the retrovirus used to transduce the cells and fibronectin, a fibronectin fragment, or a mixture thereof.
A method for obtaining a transduced population of viable cells by a retrovirus, comprising:
- infecting the cells with a retrovirus in the presence of an effective immobilized amount of fibronectin, a fibronectin fragment, or a mixture thereof to increase the efficiency of transduction of the cells by the retrovirus, said fibronectin, fibronectin fragment, or mixture thereof 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, said infecting being conducted in a medium essentially free from a polycationic agent which increases the efficiency of transduction of the cells by the retrovirus in coculture, but which polycationic agent reduces the efficiency of transduction of the cells by the retrovirus in the presence of said fibronectin, fibronectin fragment, or mixture thereof, said infecting forming a population of viable cells transduced at an efficiency greater than that which would be achieved in the presence of said polycationic agent.
A method for transducing T cells in vitro with a retrovirus, comprising infecting the cells in vitro with the retrovirus in the presence of fibronectin, a fibronectin fragment, or a mixture thereof, wherein said fibronectin or said fibronectin fragment includes a ligand which binds to the T cells and a ligand which binds to the retrovirus, so as to co-localize the retrovirus in the cells and increase the transduction efficiency of the cells.
Dependencies are inferred from the claim text.