Methods and compositions for generating an immune response by inducing cd40 and pattern recognition receptor adapters

Abstract

Provided are methods for activating an antigen-presenting cell and eliciting an immune response by inducing an inducible pattern recognition receptor adapter, or adapter fragment, and CD40 activity. Also provided are nucleic acid compositions comprising sequences coding for chimeric proteins that include an inducible CD40 peptide and an inducible pattern recognition receptor adapter or adapter fragment.

1 . A method for activating an antigen-presenting cell, which comprises:
 transfecting or transducing an antigen-presenting cell with a nucleic acid having a nucleotide sequence that encodes a chimeric protein, wherein the chimeric protein comprises (i) a membrane targeting region, (ii) a ligand-binding region (iii) a cytoplasmic CD40 polypeptide region, and (iv) a peptide selected from the group consisting of a MyD88 peptide, a truncated MyD88 peptide lacking the TIR domain, a NOD2 peptide, a RIG-1 peptide, and a TRIF peptide; and   contacting the antigen-presenting cell with a non-protein multimeric ligand that binds to the ligand-binding region;   whereby the antigen-presenting cell is activated.   
     
     
         2 . A method for activating an antigen-presenting cell, which comprises:
 transfecting or transducing an antigen-presenting cell with a nucleic acid having a nucleotide sequence that encodes a chimeric protein, wherein the chimeric protein comprises (i) a membrane targeting region, (ii) a ligand-binding region, and (iii) a truncated MyD88 peptide lacking the TIR domain; and   contacting the antigen-presenting cell with a non-protein multimeric ligand that binds to the ligand-binding region;   whereby the antigen-presenting cell is activated.   
     
     
         3 . The method of  claim 2 , wherein the chimeric protein further comprises a cytoplasmic CD40 polypeptide region. 
     
     
         4 . The method of  claim 1 , wherein the peptide has a peptide sequence selected from the group consisting of SEQ ID NO: 6, SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14, and SEQ ID NO: 16 or wherein the peptide is encoded by a nucleotide sequence selected from the group consisting of SEQ ID NO: 9, SEQ ID NO: 11, SEQ ID NO: 13, and SEQ ID NO: 15. 
     
     
         5 . The method of  claim 1 , wherein the truncated MyD88 has the peptide sequence of SEQ ID NO: 6. 
     
     
         6 . The method of  claim 1 , wherein the truncated MyD88 peptide is encoded by the nucleotide sequence of SEQ ID NO: 5. 
     
     
         7 . The method of  claim 1 , wherein the membrane targeting region is selected from the group consisting of myristoylation-targeting region, palmitoylation targeting region, prenylation region, and receptor transmembrane region. 
     
     
         8 . The method of  claim 1 , wherein the CD40 cytoplasmic polypeptide region has a peptide sequence of the cytoplasmic region of SEQ ID NO: 2. 
     
     
         9 . The method of  claim 1 , wherein the ligand-binding region is selected from the group consisting of FKBP ligand-binding region, cyclophilin receptor ligand-binding region, steroid receptor ligand-binding region, cyclophilin receptor ligand-binding region, and tetracycline receptor ligand-binding region. 
     
     
         10 . The method of  claim 1 , wherein the ligand-binding region comprises a Fv′Fvls sequence. 
     
     
         11 . The method of  claim 1 , wherein the ligand is a small molecule. 
     
     
         12 . The method of  claim 1 , wherein the ligand is dimeric. 
     
     
         13 . The method of  claim 12 , wherein the ligand is dimeric FK506 or a dimeric FK506 analog. 
     
     
         14 . The method of  claim 1 , wherein the nucleic acid is contained within a viral vector. 
     
     
         15 . The method of  claim 14 , wherein the viral vector is an adenoviral vector. 
     
     
         16 . The method of  claim 1 , wherein the nucleic acid is contained within a plasmid vector. 
     
     
         17 . The method of  claim 14 , wherein the antigen-presenting cell is contacted with the vector ex vivo. 
     
     
         18 . The method of  claim 1 , wherein the antigen-presenting cell is contacted with an antigen. 
     
     
         19 . The method of  claim 17 , wherein the antigen-presenting cell is transfected or transduced with the vector ex vivo and administered to a subject 
     
     
         20 . The method of  claim 14 , wherein the antigen-presenting cell is transduced or transfected with the nucleic acid in vivo. 
     
     
         21 . The method of  claim 19 , wherein an immune response is generated against the antigen. 
     
     
         22 . The method of  claim 21  wherein the immune response is a cytotoxic T-lymphocyte (CTL) immune response. 
     
     
         23 . The method of  claim 1 , wherein the antigen-presenting cell is a dendritic cell. 
     
     
         24 . The method of  claim 1 , wherein the nucleic acid comprises a promoter sequence operably linked to the polynucleotide sequence. 
     
     
         25 . The method of  claim 1 , wherein the antigen-presenting cell is activated without the addition of an adjuvant. 
     
     
         26 . A composition comprising a nucleic acid having a nucleotide sequence that encodes a chimeric protein, wherein the chimeric protein comprises (i) a membrane targeting region, (ii) a ligand-binding region (iii) a cytoplasmic CD40 polypeptide region, and (iv) a peptide selected from the group consisting of a MyD88 peptide, a truncated MyD88 peptide lacking the TIR domain, a NOD2 peptide, a RIG-1 peptide, and a TRIF peptide. 
     
     
         27 . A composition comprising a nucleic acid having a nucleotide sequence that encodes a chimeric protein, wherein the chimeric protein comprises (i) a membrane targeting region, (ii) a ligand-binding region, and (iii) a truncated MyD88 peptide lacking the TIR domain. 
     
     
         28 . The composition of  claim 27 , wherein the chimeric protein further comprises a cytoplasmic CD40 polypeptide region. 
     
     
         29 . The composition of  claim 26 , wherein the peptide has a peptide sequence selected from the group consisting of SEQ ID NO: 6, SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14, and SEQ ID NO: 16 or wherein the peptide is encoded by a nucleotide sequence selected from the group consisting of SEQ ID NO: 9, SEQ ID NO: 11, SEQ ID NO: 13, and SEQ ID NO: 15. 
     
     
         30 . The composition of  claim 26 , wherein the truncated MyD88 has the peptide sequence of SEQ ID NO: 6. 
     
     
         31 . The composition of  claim 26 , wherein the truncated MyD88 peptide is encoded by the nucleotide sequence of SEQ ID NO: 5. 
     
     
         32 . The composition of  claim 26 , wherein the membrane targeting region is selected from the group consisting of myristoylation-targeting region, palmitoylation targeting region, prenylation region, and receptor transmembrane region. 
     
     
         33 . The composition of  claim 26 , wherein the CD40 cytoplasmic polypeptide region has a peptide sequence of the cytoplasmic region of SEQ ID NO: 2. 
     
     
         34 . The composition of  claim 26 , wherein the ligand-binding region is selected from the group consisting of FKBP ligand-binding region, cyclophilin receptor ligand-binding region, steroid receptor ligand-binding region, cyclophilin receptor ligand-binding region, and tetracycline receptor ligand-binding region. 
     
     
         35 . The composition of  claim 26 , wherein the nucleic acid is contained within a viral vector. 
     
     
         36 . The composition of  claim 35 , wherein the viral vector is an adenoviral vector. 
     
     
         37 . The composition of  claim 26 , wherein the nucleic acid is contained within a plasmid vector. 
     
     
         38 . The composition of  claim 26 , wherein the nucleic acid comprises a promoter sequence operably linked to the polynucleotide sequence. 
     
     
         39 . A composition comprising a cell transduced or transfected with a nucleic acid of  claim 26 . 
     
     
         40 . The composition of  claim 26 , wherein the cell is an antigen-presenting cell. 
     
     
         41 . The composition of  claim 26 , wherein the cell is a dendritic cell. 
     
     
         42 . A method for treating a condition in a subject by enhancing an immune response, comprising administering to said subject a composition of  claim 26 . 
     
     
         43 . A method for treating a condition in a subject by enhancing an immune response, comprising administering to said subject a composition of  claim 39 . 
     
     
         44 . The method of  claim 43 , wherein the condition is a hyperproliferative disease. 
     
     
         45 . The method of  claim 43 , wherein the condition is an infectious disease.