Cell

Abstract

The present invention provides a cell which co-expresses a first chimeric antigen receptor (CAR) and second CAR at the cell surface, each CAR comprising: (i) an antigen-binding domain; (ii) a spacer (iii) a trans-membrane domain; and (iv) an endodomain wherein the antigen binding domains of the first and second CARs bind to different antigens, and wherein each of the first and second CARs is an activating CAR comprising an activating endodomain.

1 . A T cell which co-expresses a first chimeric antigen receptor (CAR) and second CAR at the cell surface, each CAR comprising:
 (i) an antigen-binding domain;   (ii) a spacer   (iii) a trans-membrane domain; and   (iv) an endodomain   wherein each of the first and second CARs is an activating CAR comprising an activating endodomain; and wherein the first CAR binds CD19 and the second CAR binds CD20.   
     
     
         2 - 5 . (canceled) 
     
     
         6 . A nucleic acid sequence encoding first and second chimeric antigen receptors (CARs) each CAR comprising:
 (i) an antigen-binding domain;   (ii) a spacer   (iii) a trans-membrane domain; and   (iv) an endodomain;   wherein each of the first and second CARs is an activating CAR comprising an activating endodomain; and wherein the first CAR binds CD19 and the second CAR binds CD20.   
     
     
         7 . A nucleic acid sequence according to  claim 6 , which has the following structure:
 Ag B1-spacer1-TM1-endo1-coexpr-AbB2-spacer2-TM2-endo2   in which   AgB1 is a nucleic acid sequence encoding the antigen-binding domain of the first CAR;   spacer 1 is a nucleic acid sequence encoding the spacer of the first CAR;   TM1 is a a nucleic acid sequence encoding the transmembrane domain of the first CAR;   endo 1 is a nucleic acid sequence encoding the endodomain of the first CAR;   coexpr is a nucleic acid sequence enabling co-expression of both CARs   AgB2 is a nucleic acid sequence encoding the antigen-binding domain of the second CAR;   spacer 2 is a nucleic acid sequence encoding the spacer of the second CAR;   TM2 is a a nucleic acid sequence encoding the transmembrane domain of the second CAR;   endo 2 is a nucleic acid sequence encoding the endodomain of the second CAR;   which nucleic acid sequence, when expressed in a T cell, encodes a polypeptide which is cleaved at the cleavage site such that the first and second CARs are co-expressed at the T cell surface.   
     
     
         8 . A nucleic acid sequence according to  claim 7 , wherein coexpr encodes a sequence comprising a self-cleaving peptide. 
     
     
         9 . A nucleic acid sequence according to  claim 7 , wherein alternative codons are used in regions of sequence encoding the same or similar amino acid sequences, in order to avoid homologous recombination. 
     
     
         10 - 12 . (canceled) 
     
     
         13 . A vector comprising a nucleic acid sequence according to  claim 6 . 
     
     
         14 . A retroviral vector or a lentiviral vector or a transposon according to  claim 13 . 
     
     
         15 . A method for making a T cell according to  claim 1 , which comprises the step of introducing into a T cell: a nucleic acid sequence according to  claim 6  or a vector comprising the nucleic acid sequence. 
     
     
         16 . A method according to  claim 15 , wherein the T cell is from a sample isolated from a subject. 
     
     
         17 . A pharmaceutical composition comprising a plurality of T cells according  claim 1 . 
     
     
         18 . A method for treating and/or preventing a disease, which comprises the step of administering a pharmaceutical composition according to  claim 17  to a subject. 
     
     
         19 . A method according to  claim 18 , which comprises the following steps:
 (i) isolation of a T cell-containing sample from a subject;   (ii) transduction or transfection of the T cells with: a nucleic acid sequence according to  claim 6 ; or a vector comprising the nucleic acid sequence; and   (iii) administering the T cells from (ii) to the subject.   
     
     
         20 . A method according to  claim 18 , wherein the disease is a cancer. 
     
     
         21 - 26 . (canceled) 
     
     
         27 . A natural killer (NK) cell which co-expresses a first chimeric antigen receptor (CAR) and second CAR at the cell surface, each CAR comprising:
 (i) an antigen-binding domain;   (ii) a spacer   (iii) a trans-membrane domain; and   (iv) an endodomain   wherein each of the first and second CARs is an activating CAR comprising an activating endodomain; and wherein the first CAR binds CD19 and the second CAR binds CD20.   
     
     
         28 . A cell composition comprising CAR expressing T cells according to  claim 1  made by transducing a blood-sample ex vivo with a nucleic acid encoding the first and second CARs. 
     
     
         29 . A cell composition comprising CAR expressing NK cells according to  claim 27  made by transducing a blood-sample ex vivo with a nucleic acid encoding the first and second CARs.