Methods for activating t cells using an inducible chimeric polypeptide

Abstract

The technology relates generally to the field of immunology and relates in part to methods for activating cells, including for example T cells and T cells that express chimeric antigen receptors, using an inducible chimeric polypeptide including CD40, MyD88, or CD40 and MyD88 polypeptides. The technology further relates in part to therapeutic methods for inducing an immune response and treating tumors in a patient.

1 - 30 . (canceled) 
     
     
         31 . A method for stimulating a T cell-mediated immune response in a subject, comprising administering:
 (a) a modified T cell comprising a nucleic acid that comprises a polynucleotide sequence encoding an inducible chimeric stimulating molecule, wherein the inducible chimeric stimulating molecule comprises:
 (i) a MyD88 polypeptide region or a truncated MyD88 polypeptide region lacking the TIR domain; 
 (ii) a CD40 cytoplasmic polypeptide region lacking the CD40 extracellular domain; and 
 (iii) an intracellular multimerizing region that comprises two or more FKBP12 regions, wherein each FKBP12 region comprises the amino acid sequence of SEQ ID NO:11, and 
   (b) an effective amount of a multimeric ligand that binds to the multimerization region to stimulate a T cell-mediated immune response in the subject.   
     
     
         32 . The method of  claim 31 , wherein the intracellular multimerizing region comprises two FKBP12 regions. 
     
     
         33 . The method of  claim 31 , wherein the inducible chimeric stimulating molecule comprises a truncated MyD88 polypeptide region lacking the TIR domain. 
     
     
         34 . The method of  claim 33 , wherein the truncated MyD88 polypeptide region comprises the amino acid sequence of SEQ ID NO: 5. 
     
     
         35 . The method of  claim 33 , wherein the CD40 cytoplasmic polypeptide region comprises the amino acid sequence of SEQ ID NO: 9. 
     
     
         36 . The method of  claim 33 , wherein the truncated MyD88 polypeptide region consists of the amino acid sequence of SEQ ID NO: 5. 
     
     
         37 . The method of  claim 31 , wherein the CD40 cytoplasmic polypeptide region consists of the amino acid sequence of SEQ ID NO: 9. 
     
     
         38 . The method of  claim 32 , wherein each FKBP12 region consists of the amino acid sequence of SEQ ID NO: 11. 
     
     
         39 . The method of  claim 31 , wherein the nucleic acid is a plasmid vector or viral vector. 
     
     
         40 . The method of  claim 31 , wherein the modified T cell is a CAR-T cell. 
     
     
         41 . The method of  claim 31 , wherein the inducible chimeric stimulating molecule comprises:
 (a) a truncated MyD88 polypeptide region lacking the TIR domain;   (b) a CD40 cytoplasmic polypeptide region lacking the CD40 extracellular domain; and   (c) an intracellular multimerizing region that comprises two FKBP12 regions, wherein each FKBP12 region comprises the amino acid sequence of SEQ ID NO:11.   
     
     
         42 . The method of  claim 41 , wherein the modified T cell is a CAR-T cell. 
     
     
         43 . The method of  claim 31 , wherein the modified T cell is a primary T cell. 
     
     
         44 . The method of  claim 31 , wherein the nucleic acid is integrated into the genome of the modified T cell. 
     
     
         45 . The method of  claim 31 , wherein the inducible chimeric stimulating molecule consists essentially of:
 (a) a truncated MyD88 polypeptide region lacking the TIR domain;   (b) a CD40 cytoplasmic polypeptide region lacking the CD40 extracellular domain; and   (c) an intracellular multimerizing region that comprises two FKBP12 regions, wherein each FKBP12 region comprises the amino acid sequence of SEQ ID NO:11.   
     
     
         46 . The method of  claim 31 , wherein the inducible chimeric stimulating molecule does not comprise a membrane targeting region. 
     
     
         47 . A method for treating a subject having a disease or condition associated with an elevated expression of a target antigen, comprising administering an effective amount of a multimeric ligand that binds to the multimerization region, wherein
 (a) the multimeric ligand binds to an inducible chimeric stimulating molecule comprising:
 (i) a MyD88 polypeptide region or a truncated MyD88 polypeptide region lacking the TIR domain, 
 (ii) a CD40 cytoplasmic polypeptide region lacking the CD40 extracellular domain; and 
 (iii) an intracellular multimerizing region that comprises two FKBP12 regions, wherein each FKBP12 region comprises the amino acid sequence of SEQ ID NO:11; 
   (b) T cells circulating in the subject express
 (i) the inducible chimeric stimulating molecule; and 
 (ii) a chimeric antigen receptor that binds to the target antigen; 
   (c) the target antigen is present on target cells circulating in the subject; and   (d) the number or concentration of target cells in the subject is reduced following administration of the multimeric ligand.   
     
     
         48 . The method of  claim 47 , wherein the inducible chimeric stimulating molecule further comprises a membrane targeting region. 
     
     
         49 . The method of  claim 47 , wherein the inducible chimeric stimulating molecule comprises:
 (a) a truncated MyD88 polypeptide region lacking the TIR domain;   (b) a CD40 cytoplasmic polypeptide region lacking the CD40 extracellular domain; and   (c) an intracellular multimerizing region that comprises two FKBP12 regions, wherein each FKBP12 region comprises the amino acid sequence of SEQ ID NO:11.   
     
     
         50 . The method of  claim 47 , wherein the inducible chimeric stimulating molecule consists essentially of:
 (a) a truncated MyD88 polypeptide region lacking the TIR domain;   (b) a CD40 cytoplasmic polypeptide region lacking the CD40 extracellular domain; and   (c) an intracellular multimerizing region that comprises two FKBP12 regions, wherein each FKBP12 region comprises the amino acid sequence of SEQ ID NO:11.