Methods and compositions for generating an immune response by inducing CD40 and pattern recognition receptor adapters
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 comprising a promoter operably linked to a polynucleotide that encodes a chimeric protein, wherein the chimeric protein comprises (i) a membrane targeting region, (ii) a multimeric ligand-binding region, (iii) a cytoplasmic CD40 polypeptide region lacking the extracellular domain, and (iv) a truncated MyD88 polypeptide lacking the TIR domain comprising the amino acid sequence SEQ ID NO: 6, wherein the chimeric protein is expressed in the antigen-presenting cell; 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 comprising a promoter operably linked to a polynucleotide that encodes a chimeric protein, wherein the chimeric protein comprises (i) a membrane targeting region, (ii) a multimeric ligand-binding region, and (iii) a region comprising a truncated MyD88 polypeptide lacking the TIR domain comprising the amino acid sequence SEQ ID NO: 6, wherein the truncated MyD88 polypeptide activates the NF-kappa B pathway, and wherein the chimeric protein is expressed in the antigen-presenting cell; 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 1 , wherein the truncated MyD88 polypeptide is encoded by the nucleotide sequence of SEQ ID NO: 5.
4. 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.
5. The method of claim 1 , wherein the CD40 cytoplasmic polypeptide region has an amino acid sequence of the cytoplasmic region of SEQ ID NO: 2.
6. The method of claim 1 , wherein the ligand-binding region comprises a Fv′Fvls sequence.
7. The method of claim 1 , wherein the ligand is a small molecule.
8. The method of claim 1 , wherein the ligand is dimeric.
9. The method of claim 8 , wherein the ligand is dimeric FK506 or a dimeric FK506 analog.
10. The method of claim 1 , wherein the nucleic acid is contained within a viral vector.
11. The method of claim 10 , wherein the viral vector is an adenoviral vector.
12. The method of claim 10 , wherein the antigen-presenting cell is contacted with the vector ex vivo.
13. The method of claim 1 , wherein the antigen-presenting cell is contacted with an antigen.
14. The method of claim 1 , wherein the antigen-presenting cell is a dendritic cell.
15. The method of claim 1 , wherein the antigen-presenting cell is activated without the addition of an adjuvant.
16. The method of claim 1 , wherein the truncated MyD88 polypeptide is encoded by a nucleotide sequence comprising the nucleotide sequence of SEQ ID NO: 5, wherein the nucleotide sequence of SEQ ID NO: 5 does not include SalI linkers.
17. The method of claim 13 , wherein the antigen is a prostate specific membrane antigen.
18. The method of claim 1 , wherein the ligand is AP1903 or AP20187.