CELL
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 - 28 . (canceled)
29 . A nucleic acid sequence which comprises:
a) a first nucleotide sequence encoding a first chimeric antigen receptor (CAR);
b) a second nucleotide sequence encoding a second CAR;
c) a sequence encoding a self-cleaving peptide positioned between the first and second nucleotide sequences, such that the two CARs are expressed as separate entities.
30 . The nucleic acid sequence according to claim 1 , wherein alternative codons are used in one or more portion(s) of the first and second nucleotide sequences in regions which encode the same or similar amino acid sequence(s).
31 . A vector comprising a nucleic acid according to claim 29 or 30 .
32 . The vector according to claim 31 which is a retroviral vector, a lentiviral vector or a transposon.
33 . A cell which comprises a nucleic acid sequence according to claim 29 or 30 .
34 . The cell according to claim 33 , which is a T-cell or NK cell.
35 . A method for making a cell, which comprises the step of introducing into the cell ex vivo a nucleic acid sequence or a vector comprising:
a) a first nucleotide sequence encoding a first chimeric antigen receptor (CAR),
b) a second nucleotide sequence encoding a second CAR, and
c) a sequence encoding a self-cleaving peptide positioned between the first and second nucleotide sequences, such that the two CARs are expressed as separate entities.
36 . The method of claim 35 wherein alternative codons are used in one or more portion(s) of the first and second nucleotide sequences in regions which encode the same or similar amino acid sequence(s).
37 . The method of claim 35 wherein the vector is a retroviral vector, a lentiviral vector or a transposon.
38 . The method of claim 35 wherein the cell is a T-cell or NK cell.
39 . A method according to claim 35 , wherein the cell is from a sample isolated from a subject.
40 . A pharmaceutical composition comprising a plurality of cells according to claim 33 .
41 . A pharmaceutical composition comprising a plurality of cells according to claim 34 .
42 . A method for treating a disease, which comprises the step of administering a pharmaceutical composition according to claim 40 to a subject.
43 . A method for treating a disease, which comprises the step of administering a pharmaceutical composition according to claim 41 to a subject.
44 . A method for treating a disease, which comprises the following steps:
a) isolation of a T cell-containing sample from a subject;
b) transduction or transfection of the T cells with a nucleic acid sequence or a vector comprising:
i) a first nucleotide sequence encoding a first chimeric antigen receptor (CAR),
ii) a second nucleotide sequence encoding a second CAR, and
iii) a sequence encoding a self-cleaving peptide positioned between the first and second nucleotide sequences, such that the two CARs are expressed as separate entities; and
c) administering the T cells from b) to a the subject.
45 . The method according to claim 43 , wherein the disease is a cancer.
46 . The method according to claim 44 , wherein the disease is a cancer.