Hbv epitope reactive exogenous t cell receptor (tcr) and uses thereof

Abstract

There is provided at least one isolated cell comprising at least one HBV epitope-reactive exogenous T cell receptor and/or fragment thereof, and methods for producing them. In particular, there is provided polynucleotides, constructs and vectors encoding at least one HBV epitope-reactive exogenous T cell receptor for use in the treatment of Hepatitis B Virus (HBV) and Hepatocellular Carcinoma (HCC). The invention further provides kits and methods of detection of HBV and HCC.

1 . An isolated cell comprising at least one of an HBV epitope-reactive exogenous T cell receptor and a fragment thereof, wherein the HBV epitope is HLA-A2 restricted. 
     
     
         2 . The cell according to  claim 1 , wherein the HBV epitope-reactive exogenous T cell receptor reacts with an HBV epitope that comprises at least one of: (i) at least one hepatitis B core antigen, (ii) at least one hepatitis envelope antigen, and (iii) at least one mutant of (i) or (ii). 
     
     
         3 . The cell according to  claim 1 , wherein the HBV epitope comprises at least one of: (i) an HBc18-27 epitope, (ii) an HBs370-79 epitope, and (iii) a mutant of (i) or (ii). 
     
     
         4 . The cell according to  claim 3 , wherein the HBc18-27 epitope comprises at least one of SEQ ID NO:25 and SEQ ID NO:26, and wherein the HBs370-79 epitope comprises a sequence selected from the group consisting of SEQ ID NO: 56, SEQ ID NO: 57 and SEQ ID NO: 58. 
     
     
         5 . The cell according to  claim 1 , wherein at least one of:
 (I) the HBV epitope reactive exogenous T cell receptor comprises at least one α-chain comprising a sequence that is selected from:
 (a) SEQ ID NO:9, SEQ ID NO:10 and SEQ ID NO:11, or a mutant thereof, and 
 (b) SEQ ID NO:44, SEQ ID NO:45 and SEQ ID NO:46, or a mutant thereof and 
   (II) the HBV epitope-reactive exogenous T cell receptor comprises at least one β-chain comprising a sequence that is selected from:
 (a) SEQ ID NO:21, SEQ ID NO:22 and SEQ ID NO:23 or a mutant thereof, and 
 (b) SEQ ID NO:52, SEQ ID NO:53 and SEQ ID NO:54 or a mutant thereof. 
   
     
     
         6 . The cell according to  claim 1 , wherein the exogenous T cell receptor comprises either or both of:
 (a) at least one of (i) at least one α-chain having at least 90% amino acid identity to SEQ ID NO: 12 or a fragment thereof, and (ii) at least one β-chain having at least 90% amino acid identity to SEQ ID NO:24 or a fragment thereof and   (b) at least one of (i) at least one α-chain having at least 90% amino acid identity to SEQ ID NO: 47 or a fragment thereof, and (ii) at least one β-chain having at least 90% amino acid identity to SEQ ID NO:55 or a fragment thereof.   
     
     
         7 . An isolated polynucleotide comprising at least one sequence encoding at least one of a T cell receptor α-chain and a T cell receptor β-chain, wherein the α-chain and the β-chain are of at least one HBV epitope-reactive exogenous T cell receptor, and wherein the HBV epitope is HLA-A2 restricted. 
     
     
         8 . The isolated polynucleotide according to  claim 7 , wherein at least one of: (i) the sequence encoding the α-chain comprises at least one sequence selected from SEQ ID NO:1 and SEQ ID NO:5, at least one sequence selected from SEQ ID NO:2 and SEQ ID NO:6, and at least one sequence selected from SEQ ID NO:3 and SEQ ID NO:7, and (ii) the sequence encoding the β-chain comprises at least one sequence selected from SEQ ID NO:13 and SEQ ID NO:17, at least one sequence selected from SEQ ID NO:14 and SEQ ID NO:18, and at least one sequence selected from SEQ ID NO:15 and SEQ ID NO:19. 
     
     
         9 . The isolated polynucleotide according to  claim 7 , wherein at least one of: (i) the sequence encoding the α-chain comprises SEQ ID NO:5, SEQ ID NO:6 and SEQ ID NO:7, and (ii) the sequence encoding the β-chain comprises SEQ ID NO:17, SEQ ID NO:18 and SEQ ID NO:19. 
     
     
         10 . The isolated polynucleotide according to  claim 7 , wherein, the α-chain has at least 90% sequence identity to SEQ ID NO:4 or SEQ ID NO:8, and the β-chain has at least 90% sequence identity to SEQ ID NO:16 or SEQ ID NO:20. 
     
     
         11 . The isolated polynucleotide according to  claim 7 , wherein at least one of: (i) the sequence encoding the α-chain is selected from the group consisting of SEQ ID NO:4 and SEQ ID NO:8, and (ii) the sequence encoding the β-chain is selected from the group consisting of SEQ ID NO:16 and SEQ ID NO:20. 
     
     
         12 . The isolated polynucleotide according to  claim 7 , wherein at least one of: (i) the sequence encoding the α-chain comprises a sequence selected from SEQ ID NO:40, SEQ ID NO:41 and SEQ ID NO:42, and (ii) the sequence encoding the β-chain comprises a sequence selected from SEQ ID NO:48, SEQ ID NO:49 and SEQ ID NO:50. 
     
     
         13 . The isolated polynucleotide according to  claim 7 , wherein: the α-chain of the HBV epitope-reactive exogenous T cell receptor has at least 90% sequence identity to SEQ ID NO:43, and the β-chain of the HBV epitope-reactive exogenous T cell receptor has at least 90% sequence identity to SEQ ID NO:51. 
     
     
         14 . An isolated polypeptide comprising one or more of:
 (a) at least one of (i) an α-chain comprising SEQ ID NO:9, SEQ ID NO:10 and SEQ ID NO:11, and (ii) a β-chain comprising SEQ ID NO:21, SEQ ID NO:22 and SEQ ID NO:23; and   (b) at least one of (i) an α-chain comprising SEQ ID NO:44, SEQ ID NO:45 and SEQ ID NO:46, and (ii) a β-chain comprising SEQ ID NO:52, SEQ ID NO:53 and SEQ ID NO:54;   wherein the polypeptide is at least one HBV epitope-reactive exogenous T cell receptor and the HBV epitope is HLA-A2 restricted.   
     
     
         15 . The isolated polypeptide according to  claim 14  wherein at least one of:
 (a) the α-chain has at least 90% amino acid identity to SEQ ID NO:12 and the β-chain has at least 90% amino acid identity to SEQ ID NO:24; and 
 (b) the α-chain has at least 90% amino acid identity to SEQ ID NO:47 and the β-chain has at least 90% amino acid identity to SEQ ID NO:55. 
 
     
     
         16 . The polypeptide according to  claim 14 , wherein the HBV epitope is at least one of (i) an HBc18-27 epitope, (ii) an HBs370-79 epitope, and (iii) a mutant of (i) or (ii). 
     
     
         17 . The polypeptide according to  claim 14 , wherein the polypeptide is at least one soluble T cell receptor (TCR) or a fragment thereof and is linked to at least one anti-viral drug. 
     
     
         18 . A method for treating HBV or HBV-related hepatocellular carcinoma or for inhibiting reactivation of HBV in at least one subject, comprising administering to the subject at least one immunotherapeutically effective amount of cells comprising at least one of an HBV epitope-reactive exogenous T cell receptor and a fragment thereof, wherein the HBV epitope is HLA-A2 restricted. 
     
     
         19 . An in vitro method for diagnosing at least one subject as being able to resolve HBV infection or as having or being at risk for having HBV infection or HBV related hepatocellular carcinoma, the method comprising:
 (a) providing at least one sample from at least one subject; and   (b) detecting in the sample presence or absence of at least one polynucleotide according to  claim 7 ,   wherein the presence of the polynucleotide is indicative of the subject being able to resolve the HBV infection and the absence of the polynucleotide correlates with likelihood that the subject has or is at risk for having HBV-infection or HBV-related hepatocellular carcinoma.   
     
     
         20 . An in vitro method for diagnosing at least one subject as being able to resolve HBV infection or as having or being at risk for having HBV infection or HBV related hepatocellular carcinoma, the method comprising:
 (a) providing at least one sample from at least one subject; and   (b) detecting in the sample presence or absence of at least one polypeptide according to  claim 14 ,   wherein the presence of the polypeptide is indicative of the subject being able to resolve the HBV infection and the absence of the polypeptide correlates with likelihood that the subject has or is at risk for having HBV-infection or HBV-related hepatocellular carcinoma.   
     
     
         21 . A method according to  claim 18 , wherein the HBV epitope comprises at least one of: (i) an HBc18-27 epitope, (ii) an HBs370-79 epitope, and (iii) a mutant of (i) or (ii). 
     
     
         22 . A method according to  claim 18 , wherein the HBc18-27 epitope comprises at least one of SEQ ID NO:25 and SEQ ID NO:26, and wherein the HBs370-79 epitope comprises a sequence selected from the group consisting of SEQ ID NO: 56, SEQ ID NO: 57 and SEQ ID NO: 58. 
     
     
         23 . A method according to  claim 18 , wherein at least one of:
 (I) the HBV epitope reactive exogenous T cell receptor comprises at least one α-chain comprising a sequence that is selected from:
 (a) SEQ ID NO:9, SEQ ID NO:10 and SEQ ID NO:11, or a mutant thereof, and 
 (b) SEQ ID NO:44, SEQ ID NO:45 and SEQ ID NO:46, or a mutant thereof; and 
   (II) the HBV epitope-reactive exogenous T cell receptor comprises at least one β-chain comprising a sequence that is selected from:
 (a) SEQ ID NO:21, SEQ ID NO:22 and SEQ ID NO:23 or a mutant thereof, and 
 (b) SEQ ID NO:52, SEQ ID NO:53 and SEQ ID NO:54 or a mutant thereof. 
   
     
     
         24 . A method according to  claim 18 , wherein the exogenous T cell receptor comprises either or both of:
 (a) at least one of (i) at least one α-chain having at least 90% amino acid identity to SEQ ID NO: 12 or a fragment thereof, and (ii) at least one β-chain having at least 90% amino acid identity to SEQ ID NO:24 or a fragment thereof; and   (b) at least one of (i) at least one α-chain having at least 90% amino acid identity to SEQ ID NO: 47 or a fragment thereof, and (ii) at least one β-chain having at least 90% amino acid identity to SEQ ID NO:55 or a fragment thereof.