T cell which co-expresses a CD19 chimeric antigen receptor and a CD22 chimeric antigen receptor

1. A T cell which expresses a nucleic acid comprising a nucleic acid sequence encoding both a first chimeric antigen receptor (CAR) and a second CAR as separate molecules at the cell surface, each CAR comprising: an antigen-binding domain; a spacer; a trans-membrane domain; and an endodomain,

wherein the antigen-binding domain of the first CAR binds to CD19 and the antigen-binding domain of the second CAR binds to CD22,

wherein each CAR comprises a CD3 zeta endodomain.

2. The T cell according to claim 1 , wherein the nucleic acid comprises a nucleic acid sequence which has the following structure:

AgB1-spacer1-TM1-endo1-coexpr-AgB2-spacer2-TM2-endo2

in which

AgB1 is a nucleic acid sequence encoding the antigen-binding domain of the first CAR;

spacer1 is a nucleic acid sequence encoding the spacer of the first CAR;

TM1 is a nucleic acid sequence encoding the transmembrane domain of the first CAR;

endo1 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;

spacer2 is a nucleic acid sequence encoding the spacer of the second CAR;

TM2 is a nucleic acid sequence encoding the transmembrane domain of the second CAR; and

endo2 is a nucleic acid sequence encoding the endodomain of the second CAR;

which nucleic acid sequence, when expressed in the T cell, encodes a polypeptide which is cleaved at a cleavage site such that the first and second CARs are co-expressed at the T cell surface.

3. The T cell according to claim 2 , wherein coexpr comprises a nucleic acid sequence encoding a self-cleaving peptide.

4. The T cell according to claim 2 , wherein alternative codons are used in regions of sequence encoding the same amino acid sequences, in order to avoid homologous recombination.

5. A pharmaceutical composition comprising a plurality of T cells according to claim 2 .

6. The T cell according to claim 1 , wherein each endodomain is a compound endodomain comprised of (a) the CD3 zeta domain and (b) a co-stimulatory domain or a TNF receptor family endodomain.

7. The T cell according to claim 2 , wherein the endodomain of the first CAR and the endodomain of the second CAR each is a compound endodomain comprising (a) the CD3 zeta domain in combination with (b) a co-stimulatory domain or a TNF receptor family endodomain.

8. A pharmaceutical composition comprising a plurality of T cells according to claim 1 .

9. A method of disease treatment, which comprises a step of administering a pharmaceutical composition according to claim 8 to a subject.

10. The method of claim 9 , wherein the disease is a B cell malignancy.

11. A method of disease treatment, which comprises a step of administering to a subject a pharmaceutical composition comprising a plurality of T cells which co-express a first chimeric antigen receptor (CAR) and second CAR as separate molecules at the cell surface, each CAR comprising:

an antigen-binding domain; a spacer; a trans-membrane domain; and an endodomain,

wherein the antigen-binding domain of the first CAR binds to CD19 and the antigen-binding domain of the second CAR binds to CD22, and

wherein each CAR comprises a CD3 zeta endodomain.

12. The method according to claim 11 , wherein the spacer of the first CAR is different from the spacer of the second CAR.

13. A method according to claim 11 , which comprises the following steps:

(i) isolation of a T cell-containing sample from a subject;

(ii) transduction or transfection of T cells from the sample with:

(a) a nucleic acid or vector comprising a nucleic acid sequence encoding both the first and second CARs; or

(b) a first nucleic acid or vector and a second nucleic acid or vector, wherein

(1) the first nucleic acid or vector comprises a nucleic acid sequence encoding the first CAR and comprises the following structure:

AgB1-spacer1-TM1-endo1

in which

AgB1 is a nucleic acid sequence encoding the antigen-binding domain of the first CAR;

spacer1 is a nucleic acid sequence encoding the spacer of the first CAR;

TM1 is a nucleic acid sequence encoding the transmembrane domain of the first CAR; and

endo1 is a nucleic acid sequence encoding the endodomain of the first CAR; and

(2) the second nucleic acid or vector comprises a nucleic acid sequence encoding the second CAR and comprises the following structure:

AgB2-spacer2-TM2-endo2

in which

AgB2 is a nucleic acid sequence encoding the antigen-binding domain of the second CAR;

spacer2 is a nucleic acid sequence encoding the spacer of the second CAR;

TM2 is a nucleic acid sequence encoding the transmembrane domain of the second CAR; and

endo2 is a nucleic acid sequence encoding the endodomain of the second CAR; and

(iii) administering the T cells from (ii) to the subject.

14. A method according to claim 11 , wherein the disease is a cancer.

15. A method according to claim 14 , wherein the cancer is B cell malignancy.

16. The method according to claim 11 , wherein each CAR has a compound endodomain comprised of (a) the CD3 zeta domain and (b) a co-stimulatory domain or a TNF receptor family endodomain.