Dna vaccines and methods for the prevention of transplantation rejection

Abstract

Methods for preventing, delaying the onset of, or treating rejection of an allograft using a DNA vaccine, where the vaccine can comprise a polynucleotide encoding a pro-apoptotic protein, like BAX and/or a polynucleotide encoding an autoantigen or donor antigen, like secreted glutamic acid decarboxylase 55. Administering one of the DNA vaccines to a transplant recipient, as described herein, can induce a donor specific tolerogenic response.

1 . A method for preventing, delaying the onset of, or treating rejection of an allograft or an allogeneic transplant from a donor to a recipient, comprising:
 administering to the recipient one or more than one dose of a DNA vaccine selected from the group consisting of:   (1) a first plasmid comprising
 (a) a polynucleotide encoding a pro-apoptotic protein; 
 (b) a promoter controlling the expression of the polynucleotide encoding the pro-apoptotic protein; 
   (2) a second plasmid comprising:
 (a) a polynucleotide sequence encoding an autoantigen or donor antigen; 
 (b) a promoter controlling expression of the polynucleotide sequence encoding the autoantigen or donor antigen; and 
 (c) a plurality of CpG motifs, where the CpG motifs are methylated sufficiently to diminish the recipient's immune response to unmethylated CpG motifs; and 
   (3) a third plasmid comprising:
 (a) a polynucleotide sequence encoding an autoantigen or donor antigen; 
 (b) a promoter controlling expression of the polynucleotide sequence encoding the autoantigen or donor antigen; 
 (c) a polynucleotide encoding a pro-apoptotic protein; 
 (d) a promoter controlling expression of the polynucleotide encoding the pro-apoptotic protein; and 
 (e) a plurality of CpG motifs, where the CpG motifs are methylated sufficiently to diminish the recipient's immune response to unmethylated CpG motifs; 
   
     
     
         2 . The method of  claim 1 , wherein engrafted tissues or transplanted organs are selected from the group consisting of skin grafts, islet cell transplants, and partial or whole organ transplants. 
     
     
         3 . The method of  claim 2 , wherein the partial or whole organ transplants are selected from the group consisting of hearts, lungs, kidneys and livers. 
     
     
         4 . The method of  claim 1 , where the DNA vaccine is a plasmid comprising a plurality of methylated CpG motifs, where the plasmid is resistant to digestion by the restriction enzyme HpαII. 
     
     
         5 . The method of  claim 1 , where the DNA vaccine is a plasmid comprising a plurality of CpG motifs, where the CpG motifs of the plasmid are methylated by SssI methylase. 
     
     
         6 . The method of  claim 1 , where the promoter capable of expressing the polynucleotide encoding the autoantigen or the donor antigen, or the promoter capable of expressing the polynucleotide encoding the pro-apoptotic protein, or both the promoter capable of expressing the polynucleotide encoding the autoantigen or the donor antigen, and the promoter capable of expressing the polynucleotide encoding the pro-apoptotic protein maintain their promoter function after methylation. 
     
     
         7 . The method of  claim 1 , where the third plasmid further comprises an internal ribosome entry site (IRES) sequence to permit translation of the polynucleotide encoding the autoantigen or the donor antigen, and the polynucleotide encoding the pro-apoptotic protein from the same transcript. 
     
     
         8 - 10 . (canceled) 
     
     
         11 . The method of  claim 1 , where the autoantigen or donor antigen is selected from the group consisting of carbonic anhydrase II, collagen, CYP2D6 (cytochrome P450, family 2, subfamily Device 400, polypeptide 6), glutamic acid decarboxylase, secreted glutamic acid decarboxylase 55, SEQ ID NO:1, insulin, myelin basic protein and SOX-10 (SRY-box containing gene 10). 
     
     
         12 . The method of  claim 1 , where the pro-apoptotic protein is selected from the group consisting of BAX, SEQ ID NO: 2  NO:2, a modified caspase, Tumor Necrosis Factor Receptor, Death Receptor 3 (DR3), Death Receptor 4 (DR4), Death Receptor 5 (DRS) and a FAS receptor. 
     
     
         13 . The method of  claim 7 , where the internal ribosome entry site (IRES) sequence is SEQ ID NO:3 from the EMC virus. 
     
     
         14 . The method of  claim 1 , where the DNA vaccine is administered in an effective dose, wherein an effective dose is an amount sufficient to prevent, delay the onset or treat rejection of an allograft or an allogeneic transplant by the recipient. 
     
     
         15 . The method of  claim 1 , where the DNA vaccine is administered in an effective dose, wherein an effective dose is an amount sufficient to induce a donor-specific tolerogenic response. 
     
     
         16 . The method of  claim 1 , where the DNA vaccine is administered in a plurality of doses. 
     
     
         17 . The method of  claim 1 , where the dose of the DNA vaccine is about 0.001 mg/Kg of body weight of the recipient to about 100 mg/Kg of body weight of the recipient. 
     
     
         18 . The method of  claim 1 , where the dose is administered weekly between two times and 100 times. 
     
     
         19 . The method of  claim 1 , where the DNA vaccine is administered by an epidermal, intradermal, intramuscular, intranasal, intravenous, intraperitoneal or oral route. 
     
     
         20 . The method of  claim 1 , where the DNA vaccine is administered by an injection proximal to the site of the allograft or allogeneic transplant. 
     
     
         21 . The method of  claim 1 , further comprising administering a dose of one or more than one immunosuppressant agent before, on the day of, and/or after engraftment or transplantation. 
     
     
         22 . The method of  claim 21 , wherein the dose of one or more than one immunosuppressant agent is administered simultaneously, separately or sequentially. 
     
     
         23 . The method of  claim 21 , wherein the one or more than one immunosuppressant agent is selected from the group consisting of corticosteroids, glucocorticoids,-cyclophosphamide, 6-mercaptopurine, azathioprine, methotrexate cyclosporine, mycophenolate mofetil, mycophenolic acid, tacrolimus, sirolimus, everolimus, mizoribine, leflunomide, deoxyspergualin, brequinar, azodicarbonamide, vitamin D analogs, antilymphocyte globulin, antithymocyte globulin, anti-CD3 monoclonal antibodies, anti- interleukin-2 receptor (anti-CD25) antibodies, anti-CD52 antibodies, anti-CD20 antibodies, anti-tumor necrosis factor reagents, and LFA-1 inhibitors. 
     
     
         24 . The method of  claim 21 , comprising administering a single dose of antilymphocyte globulin, at a dosage of about 1.6 mg/20 g of body weight, on the day of engraftment or transplantation. 
     
     
         25 . The method of  claim 21 , comprising administering rapamycin at a dosage of from 0.05 to 15 mg/day. 
     
     
         26 . (canceled) 
     
     
         27 . (canceled) 
     
     
         28 . The method of  claim 1 , wherein administration of the plasmid further induces a donor-specific tolerogenic response. 
     
     
         29 . The method of  claim 1 , wherein administration of the plasmid elevates expression of inhibitory Fc receptor, FcγIIb, and Il-1 antagonist Il 1RA cytokine. 
     
     
         30 . The method of  claim 1 , wherein administration of the plasmid reduces an autoimmune response. 
     
     
         31 . The method of  claim 1 , wherein administration of the plasmid reduces expression of Tnfα (tumor necrosis factor) and Ifny (gamma interferon). 
     
     
         32 . (canceled) 
     
     
         33 . The method of  claim 1 , wherein administration of the plasmid elevates expression of IL-10 (interleukin 10). 
     
     
         34 . The method of  claim 1 , wherein administration of the plasmid elevates expression of TGFβ1 (transforming growth factor (β1). 
     
     
         35 . The method of  claim 1 , wherein administration of the plasmid increases regulatory T cell activity. 
     
     
         36 . The method of  claim 1 , further comprising selecting a recipient in need of a graft or transplant and an allograft or allogeneic transplant donor, and grafting tissue or transplanting a solid organ from the donor to the recipient; 
     
     
         37 . A method for inducing a tolerogenic response to a graft or transplant from a donor to a recipient, comprising administering to the recipient one or more than one dose of a DNA vaccine selected from the group consisting of:
 1. a first plasmid comprising
 a) a polynucleotide encoding a pro-apoptotic protein; 
 b) a promoter controlling the expression of the polynucleotide encoding the pro-apoptotic protein; 
   2 .a second plasmid comprising:
 a) a polynucleotide sequence encoding an autoantigen or donor antigen; 
 b) a promoter controlling expression of the polynucleotide sequence encoding the autoantigen or donor antigen; and 
 c) a plurality of CpG motifs, where the CpG motifs are methylated sufficiently to diminish the recipient's immune response to unmethylated CpG motifs; and 
   3. a third plasmid comprising:
 a) a polynucleotide sequence encoding an autoantigen or donor antigen; 
 b) a promoter controlling expression of the polynucleotide sequence encoding the autoantigen or donor antigen; 
 c) a polynucleotide encoding a pro-apoptotic protein; and 
 d) a promoter controlling expression of the polynucleotide encoding the pro-apoptotic protein; and 
 e) a plurality of CpG motifs, where the CpG motifs are methylated sufficiently to diminish the recipient's immune response to unmethylated CpG motifs. 
   
     
     
         38 . A method for preventing or reducing graft or transplant-induced autoimmunity comprising, administering to a graft or transplant recipient one or more than one dose of a DNA vaccine selected from the group consisting of:
 1. a first plasmid comprising
 a) a polynucleotide encoding a pro-apoptotic protein; 
 b) a promoter controlling the expression of the polynucleotide encoding the pro-apoptotic protein; 
   2. a second plasmid comprising:
 a) a polynucleotide sequence encoding an autoantigen or donor antigen; 
 b) a promoter controlling expression of the polynucleotide sequence encoding the autoantigen or donor antigen; and 
 c) a plurality of CpG motifs, where the CpG motifs are methylated sufficiently to diminish the recipient's immune response to unmethylated CpG motifs; and 
   3. a third plasmid comprising:
 a) a polynucleotide sequence encoding an autoantigen or donor antigen; 
 b) a promoter controlling expression of the polynucleotide sequence encoding the autoantigen or donor antigen; 
 c) a polynucleotide encoding a pro-apoptotic protein; and 
 d) a promoter controlling expression of the polynucleotide encoding the pro-apoptotic protein; and 
 e) a plurality of CpG motifs, where the CpG motifs are methylated sufficiently to diminish the recipient's immune response to unmethylated CpG motifs.