Method for genetic immunization and introduction of molecules into skeletal muscle and immune cells

Abstract

A method is disclosed for enhanced vaccination and genetic vaccination of mammals. The vaccination is accomplished by delivering molecules such as proteins and nucleic acids into skeletal muscle and other cells residing in the skeletal muscle in vivo. The protein or nucleic acid is first injected into the muscle at one or multiple sites. Immediately or shortly after injection, electrodes are placed flanking the injection site and a specific amount of electrical current is passed through the muscle. The electrical current makes the muscle permeable, thus allowing the pharmaceutical drug or nucleic acid to enter the cell. The efficiency of transfer permits robust immune responses using DNA vaccines and produces sufficient secreted proteins for systemic biological activity to be observed.

We claim:  
     
         1 . A method of delivering a molecule to the skeletal muscle of a mammal in vivo comprising: 
 injecting a local anesthetic into the skeletal muscle of the mammal;    injecting a molecule into an injection site in the skeletal muscle;    positioning electrodes near the injection site such that current traveling through the electrodes passes through the injection site; and    electrically stimulating the muscle with an electrical current having a field strength of from about 10 V/cm to about 300 V/cm.    
     
     
         2 . The method of  claim 1 , wherein the electrical current has a field strength of from about 10 V/cm to about 233 V/cm.  
     
     
         3 . The method of  claim 2 , wherein the electrical current is delivered as one or more square bipolar pulses.  
     
     
         4 . The method of  claim 1 , wherein the electrical current has a field strength of from about 12 V/cm to about 175 V/cm.  
     
     
         5 . The method of  claim 1 , wherein the electrical current has a field strength of from about 125 V/cm to about 233 V/cm.  
     
     
         6 . The method of  claim 1 , wherein the electrical current is delivered as one or more square bipolar pulses.  
     
     
         7 . The method of  claim 1 , wherein the local anesthetic is Marcain.  
     
     
         8 . The method of  claim 7 , wherein the electrical current has a field strength of from about 10 V/cm to about 233 V/cm.  
     
     
         9 . The method of  claim 7 , wherein the electrical current has a field strength of from about 12 V/cm to about 175 V/cm.  
     
     
         10 . The method of  claim 7 , wherein the electrical current has a field strength of from about 125 V/cm to about 233 V/cm.  
     
     
         11 . The method of  claim 7 , wherein the electrical current is delivered as one or more square bipolar pulses.  
     
     
         12 . The method of  claim 1 , wherein the molecule is an antigen.  
     
     
         13 . The method of  claim 12 , wherein the antigen is a protein.  
     
     
         14 . The method of  claim 1 , wherein the molecule in a nucleic acid molecule.  
     
     
         15 . The method of  claim 14 , wherein the nucleic acid molecule is a DNA molecule.