Stimulation of cell-mediated immune responses by targeted particulate genetic immunization

Abstract

The present invention relates to various methods of genetic immunization for the purpose of providing antigen-specific immunity in a mammalian host, including a human host. The invention is based on the ability to direct particulate polynucleotides which express an antigenic protein or protein fragment to the cytoplasm of host target cells, such as antigen presenting cells. A directed delivery of such particulate polynucleotides to the cytoplasm of antigen presenting cells will stimulate antigen-specific CTL production, thus promoting destruction of affected cells such as neoplastic cells and virally infected cells.

What is claimed is:  
     
         1 . An in vivo method of therapeutic or prophylactic genetic immunization of a mammalian host, which comprises: 
 (a) generating a DNA fragment which expresses an antigenic protein or antigenic protein fragment;    (b) distributing said DNA fragment on a particle surface, resulting in a particulate polynucleotide;    (c) inoculating said mammalian host with said particulate polynucleotide; and,    (d) delivering said particulate polynucleotide to the cytoplasm of a target cell within said mammalian host, such that said expressed antigenic protein or antigenic protein fragment is presented to the membrane surface of said target cell through the MHC class I pathway.    
     
     
         2 . The method of  claim 1  wherein said mammalian host is a human.  
     
     
         3 . The method of  claim 2  wherein said DNA fragment expresses a tumor rejection antigen, viral antigen or antigenic protein fragment thereof.  
     
     
         4 . The method of  claim 3  wherein said target cell is an antigen presenting cell.  
     
     
         5 . The method of  claim 4  wherein said antigen presenting cell resides within or migrates to the lymphoid tissue of said human host.  
     
     
         6 . The method of  claim 5  wherein said tumor rejection antigen is selected from the group consisting of MAGE-1 and MAGE 3.  
     
     
         7 . The method of  claim 5  wherein said tumor rejection antigen is Melan-A.  
     
     
         8 . The method of  claim 5  wherein said tumor rejection antigen is gp100.  
     
     
         9 . The method of  claim 5  wherein said tumor rejection antigen is p53.  
     
     
         10 . The method of  claim 5  wherein said tumor rejection antigen is CEA.  
     
     
         11 . The method of  claim 5  wherein said tumor rejection antigen is HER2/neu.  
     
     
         12 . The method of  claim 5  wherein said viral antigen is HIV gp120, HIV gp160.  
     
     
         13 . The method of  claim 5  wherein said viral antigen is Influenza virus nucleoprotein.  
     
     
         14 . The method of claim S wherein said viral antigen is Hepatitis B surface antigen.  
     
     
         15 . An in vivo method of therapeutic or prophylactic genetic immunization of a mammalian host, which comprises: 
 (a) generating a DNA fragment which expresses an antigenic protein or antigenic protein fragment;    (b) distributing said DNA fragment on a particle surface, resulting in a particulate polynucleotide;    (c) inoculating said mammalian host with said particulate polynucleotide using a biolistic device; and,    (d) delivering said particulate polynucleotide to the cytoplasm of a target cell within said mammalian host, such that said expressed antigenic protein or antigenic protein fragment is presented to the membrane surface of said target cell through the MHC class I pathway.    
     
     
         16 . The method of  claim 15  wherein said mammalian host is a human.  
     
     
         17 . The method of  claim 16  wherein said DNA fragment expresses a tumor rejection antigen, viral antigen or antigenic protein fragment thereof.  
     
     
         18 . The method of  claim 17  wherein said target cell is an antigen presenting cell.  
     
     
         19 . The method of  claim 18  wherein said antigen presenting cell resides within or migrates to the lymphoid tissue of said human host.  
     
     
         20 . The method of  claim 19  wherein said tumor rejection antigen is selected from the group consisting of MAGE-1 and MAGE 3.  
     
     
         21 . The method of  claim 19  wherein said tumor rejection antigen is Melan-A.  
     
     
         22 . The method of  claim 19  wherein said tumor rejection antigen is gp100.  
     
     
         23 . The method of  claim 19  wherein said tumor rejection antigen is p53.  
     
     
         24 . The method of  claim 19  wherein said tumor rejection antigen is CEA.  
     
     
         25 . The method of  claim 19  wherein said tumor rejection antigen is HER2/neu.  
     
     
         26 . The method of  claim 19  wherein said viral antigen is HIV gp120, HIV gp160.  
     
     
         27 . The method of  claim 19  wherein said viral antigen is Influenza virus nucleoprotein.  
     
     
         28 . The method of  claim 19  wherein said viral antigen is Hepatitis B surface antigen.  
     
     
         29 . An in vivo method of therapeutic or prophylactic genetic immunization of a mammalian host, which comprises: 
 (a) generating a DNA fragment which expresses an antigenic protein or antigenic protein fragment;    (b) distributing said DNA fragment on a particle surface, resulting in a particulate polynucleotide;    (c) inoculating said mammalian host with said particulate polynucleotide by direct injection; and,    (d) delivering said particulate polynucleotide to the cytoplasm of a target cell within said mammalian host, such that said expressed antigenic protein or antigenic protein fragment is presented to the membrane surface of said target cell through the MHC class I pathway.    
     
     
         30 . The method of  claim 29  wherein said mammalian host is a human.  
     
     
         31 . The method of  claim 30  wherein direct injection is by subcutaneous injection.  
     
     
         32 . The method of  claim 31  wherein said recombinant DNA vector fragment expresses a tumor rejection antigen, viral antigen or antigenic protein fragment thereof.  
     
     
         33 . The method of  claim 32  wherein said target cell is an antigen presenting cell.  
     
     
         34 . The method of  claim 33  wherein said antigen presenting cell resides within or migrates to the lymphoid tissue of said human host.  
     
     
         35 . The method of  claim 34  wherein said tumor rejection antigen is selected from the group consisting of MAGE-1 and MAGE 3.  
     
     
         36 . The method of  claim 34  wherein said tumor rejection antigen is Melan-A.  
     
     
         37 . The method of  claim 34  wherein said tumor rejection antigen is gp100.  
     
     
         38 . The method of  claim 34  wherein said tumor rejection antigen is p53.  
     
     
         39 . The method of  claim 34  wherein said tumor rejection antigen is CEA.  
     
     
         40 . The method of  claim 34  wherein said tumor rejection antigen is HER2/neu.  
     
     
         41 . The method of  claim 34  wherein said viral antigen is HIV gp120, HIV gp160.  
     
     
         42 . The method of  claim 34  wherein said viral antigen is Influenza virus nucleoprotein.  
     
     
         43 . The method of  claim 34  wherein said viral antigen is Hepatitis B surface antigen.  
     
     
         44 . An ex vivo method of therapeutic or prophylactic genetic immunization of a mammalian host, which comprises: 
 (a) generating a DNA fragment which expresses an antigenic protein or antigenic protein fragment;    (b) distributing said DNA fragment on a particle surface, resulting in a particulate polynucleotide;    (c) delivering said particulate polynucleotide to the cytoplasm of a target cell of a mammalian host in vitro, such that said expressed antigenic protein or antigenic protein fragment is presented on the membrane surface of said target cell through the MHC class I pathway; and,    (d) inoculating said mammalian host with said target cell by direct injection.    
     
     
         45 . The method of  claim 44  wherein said mammalian host is a human.  
     
     
         46 . The method of  claim 45  wherein direct injection is by subcutaneous injection.  
     
     
         47 . The method of  claim 46  wherein said recombinant DNA vector fragment expresses a tumor rejection antigen, viral antigen or antigenic protein fragment thereof.  
     
     
         48 . The method of  claim 47  wherein said target cell is an antigen presenting cell.  
     
     
         49 . The method of  claim 48  wherein said antigen presenting cells resides within or migrates to the lymphoid tissue of said human host.  
     
     
         50 . The method of  claim 49  wherein said tumor rejection antigen selected from the group consisting of MAGE-1 and MAGE 3.  
     
     
         51 . The method of  claim 49  wherein said tumor rejection antigen is Melan-A.  
     
     
         52 . The method of  claim 49  wherein said tumor rejection antigen is gp100.  
     
     
         53 . The method of  claim 49  wherein said tumor rejection antigen is p53.  
     
     
         54 . The method of  claim 49  wherein said tumor rejection antigen is CEA.  
     
     
         55 . The method of  claim 49  wherein said tumor rejection antigen is HER2/nue.  
     
     
         56 . The method of  claim 49  wherein said viral antigen is HIV gp120, HIV gp160.  
     
     
         57 . The method of  claim 49  wherein said viral antigen is Influenza virus nucleoprotein.  
     
     
         58 . The method of  claim 49  wherein said viral antigen is Hepatitis B surface antigen.  
     
     
         59 . An ex vivo method of therapeutic or prophylactic genetic immunization of a mammalian host, which comprises: 
 (a) generating a DNA fragment(s) which express a molecule which enhances the antigen presentation function of an APC;    (b) distributing said DNA fragment(s) on a particle surface, resulting in a particulate polynucleotide;    (c) delivering said particulate polynucleotide to the cytoplasm of a target cell of a mammalian host in vitro, such that said expressed antigen presentation enhancing protein or proteins is expressed in a biologically significant form and at biologically significant levels;    (d) inoculating said mammalian host with said target cell by direct injection.    
     
     
         60 . The method of  claim 59  wherein said mammalian host is a human.  
     
     
         61 . The method of  claim 60  wherein direct injection is by subcutaneous injection.  
     
     
         62 . The method of  claim 61  wherein said target cell is an antigen presenting cell.  
     
     
         63 . The method of  claim 62  wherein said antigen presenting cell resides within or migrates to the lymphoid tissue of said human host.  
     
     
         64 . The method of  claim 63  wherein said DNA vector fragment expresses a costimulatory molecule.  
     
     
         65 . The method of  claim 64  wherein said costimulatory molecule is selected from the group consisting of CD80 and CD86.  
     
     
         66 . The method of  claim 63  wherein said DNA vector fragment expresses a cytokine molecule  
     
     
         67 . The method of  claim 66  wherein said cytokine molecule is selected from the group consisting of IL-12, IL-4 and IL-2.