METHOD OF TREATMENT OF CANCER OR TUMOUR

Abstract

The present invention provides methods of treating, preventing or delaying the progress of cancer and/or tumour in a subject comprising administering to the subject a treatment regimen comprising an effective amount of a PD-1 axis binding antagonist and a population of modified immunoresponsive cells expressing or presenting a heterologous TCR. The invention also provides methods of enhancing immune function in a subject having cancer and/or tumour comprising administering to the subject a treatment regimen comprising an effective amount of a PD-1 axis binding antagonist and a population of modified immunoresponsive cells expressing or presenting a heterologous TCR.

Claims

1. A method of treating, preventing or delaying the progression of cancer and/or tumour in a subject comprising administering to the subject a treatment regimen comprising an effective amount of a PD-1 axis binding antagonist and a population of modified immunoresponsive cells expressing or presenting a heterologous TCR.

2. The method of claim 1, wherein the PD-1 axis binding antagonist is selected from the group consisting of (a) a PD-1 binding antagonist, (b) a PD-L1 binding antagonist, (c) a PD-L2 binding antagonist.

3. The method of claim 2, wherein: (i) the PD-1 axis binding antagonist binds to PD-1 and/or binds to SEQ ID NO: 1, optionally wherein the PD-1 binding antagonist inhibits the binding of PD-1 to its ligand binding partners, preferably wherein the PD-1 binding antagonist inhibits the binding of PD-1 to PD-L1 and/or PDL2; (ii) the PD-1 axis binding antagonist binds to PD-L1 and/or binds to SEQ ID NO: 2, optionally wherein the PD-L1 binding antagonist inhibits the binding of PD-L1 to its ligand binding partners, preferably wherein the PD-L1 binding antagonist inhibits the binding of PD-L1 to one or more of PD-1, CD80 and B7-1.

4. The method of claim 3, wherein the PD-1 or PD-L1 binding antagonist is an antibody; optionally wherein: (a) the anti-PD-L1 antibody inhibits binding between PD-L1 and PD-1 and/or between PD-L1 and B7-1, (b) the anti-PD-L1 antibody inhibits PD-L1 on the cancer cell surface from transducing a signal to the intracellular pathway, (c) the anti-PD-1 antibody inhibits binding between PD-L1 and PD-1 and/or between PD-L2 and PD-1, (d) the anti-PD-1 antibody inhibits PD-1 on the T cell surface from transducing a signal to the intracellular pathway.

5. The method of claim 4, wherein: (1) the PD-L1 binding antagonist is selected from (a) Durvalumab, Imfinzi or MEDI4736, (b) Atezolizumab, Tecentriq or MPDL3280A, (c) Avelumab, Bavencio or MSB0010718C, or (d) MDX-1105, BMS-936559, optionally wherein the PD-L1 binding antagonist is an antibody comprising: (i) a heavy chain comprising SEQ ID NO:4, and a light chain comprising SEQ ID NO:5, or variable regions thereof or CDRs thereof, (ii) a heavy chain comprising SEQ ID NO:6, and a light chain comprising SEQ ID NO:7, or variable regions thereof or CDRs thereof, (iii) a heavy chain comprising SEQ ID NO:8, and a light chain comprising SEQ ID NO:9, or variable regions thereof or CDRs thereof, or (iv) a heavy chain comprising SEQ ID NO:10, and a light chain comprising SEQ ID NO:11, or variable regions thereof or CDRs thereof, and further optionally wherein the antibody is monoclonal, human or humanised and is a full length or antigen-binding fragment thereof, Fv, Fab, Fab′, Fab′-SH, F(ab′)2; diabody; linear antibody; single-chain antibody molecule, scFv; or (2) the PD-1 binding antagonist is selected from (a) Pembrolizumab, Keytruda, Lambrolizumab or MK-3475, (b) Cemiplimab, Libtayo, or REGN-2810, or (c) BMS/ONO, Nivolumab, Opdivo, ONO-4538, BMS-936558 or MDX1106, optionally wherein the PD-1 binding antagonist is an antibody comprising: (i) a heavy chain comprising SEQ ID NO:12, and a light chain comprising SEQ ID NO:13, or variable regions thereof or CDRs thereof, (ii) a heavy chain comprising SEQ ID NO:14, and a light chain comprising SEQ ID NO:15, or variable regions thereof or CDRs thereof, or (iii) a heavy chain comprising SEQ ID NO:16, and a light chain comprising SEQ ID NO: 17, or variable regions thereof or CDRs thereof, and further optionally wherein the antibody is monoclonal, human or humanised and is a full length or antigen-binding fragment thereof, Fv, Fab, Fab′, Fab′-SH, F(ab′)2; diabody; linear antibody; single-chain antibody molecule, scFv.

6. The method of claim 2, wherein the PD-1 axis binding antagonist is a PD-L2 binding antagonist and/or binds to SEQ ID NO: 3, optionally wherein the PD-L2 binding antagonist is an antibody or an immunoadhesin.

7. The method of claim 1, wherein (a) the heterologous TCR binds or specifically binds to a cancer and/or tumour antigen or peptide antigen thereof, or (b) the heterologous TCR binds or specifically binds to a cancer and/or tumour antigen or peptide antigen thereof associated with a cancerous condition and/or presented by tumour or cancer cell or tissue; optionally wherein: (i) the cancer and/or tumour antigen or peptide antigen thereof is complexed with a peptide presenting molecule, optionally major histocompatibility complex (MHC) or human leukocyte antigen (HLA), optionally class I or class II, optionally wherein the peptide is complexed with HLA-A2 or HLA-A*02, or HLA-A*0201; (ii) the cancer and/or tumour antigen or peptide antigen thereof is a cancer-testis antigen; (iii) the cancer and/or tumour antigen or peptide antigen thereof is selected from any of; NY-ESO-1, MART-1 (melanoma antigen recognized by T cells), WT1 (Wilms tumor 1), gp100 (glycoprotein 100), tyrosinase, PRAME (preferentially expressed antigen in melanoma), p53, HPV-E6/HPV-E7 (human papillomavirus), HBV, TRAIL, DR4, Thyroglobin, TGFBII frameshift antigen, LAGE-1A, KRAS, CMV (cytomegalovirus), CEA (carcinoembryonic antigen), AFP (α-fetoprotein), MAGE-AL MAGE-A2, MAGE-A3, MAGE-A4, MAGE-A6, MAGE-A8, and MAGE-A9, MAGE-A10, or MAGE-A12, or peptide antigen thereof; (iv) the cancer and/or tumour antigen or peptide antigen is MAGE-A4 or peptide thereof, preferably the sequence GVYDGREHTV or SEQ ID NO: 18; and/or (v) the heterologous TCR binds specifically and/or selectively to the cancer and/or tumour antigen or peptide antigen thereof and/or the peptide presenting molecule and/or complex thereof.

8. The method of claim 1, wherein: (a) the heterologous TCR comprises a TCR alpha chain variable domain and a TCR beta chain variable domain, wherein: (i) the alpha chain variable domain comprises CDRs having the sequences VSPFSN (αCDR1), SEQ ID NO:27 or amino acids 48-53 of SEQ ID NO:21, LTFSEN (αCDR2), SEQ ID NO:28 or amino acids 71-76 of SEQ ID NO:21, and CVVSGGTDSWGKLQF (αCDR3), SEQ ID NO:29 or amino acids 111-125 of SEQ ID NO:21, and (ii) the beta chain variable domain comprises CDRs having the sequences KGHDR (βCDR1), SEQ ID NO:30 or amino acids 46-50 of SEQ ID NO:23, SFDVKD (βCDR2), SEQ ID NO:31 or amino acids 68-73 of SEQ ID NO:23, and CATSGQGAYEEQFF (βCDR3), SEQ ID NO:32 or amino acids 110-123 of SEQ ID NO:23; or sequences having at least 80% sequence identity thereto; and/or b) the heterologous TCR comprises a TCR in which the alpha chain variable domain comprises an amino acid sequence that has at least 80%, identity to SEQ ID NO:25 and/or the beta chain variable domain comprising an amino acid sequence that has at least 80% identity to SEQ ID NO:26.

9. The method according to claim 1, wherein the population of modified immunoresponsive cells expressing or presenting a heterologous TCR further expresses or presents a heterologous co-receptor, optionally wherein the co-receptor is a CD8 co-receptor, optionally wherein: (i) the heterologous CD8 co-receptor is a heterodimer or a homodimer, a CD8ab heterodimer or a CD8αα homodimer; and/or (ii) the heterologous CD8 co-receptor comprises; (a) a CDR 1 of at least 80% sequence identity to amino acid sequence VLLSNPTSG, SEQ ID NO:33, CDR 2 of at least 80% sequence identity to amino acid sequence YLSQNKPK SEQ ID NO:34 and CDR 3 of at least 80% sequence identity amino acid sequence LSNSIM SEQ ID NO:35, (b) a CDR 1 of amino acid sequence VLLSNPTSG, SEQ ID NO:33, CDR 2 of amino acid sequence YLSQNKPK SEQ ID NO:34 and CDR 3 of amino acid sequence LSNSIM SEQ ID NO:35, (c) an amino acid sequence having at least 80% sequence identity to amino acids number 22 to 235 of SEQ ID NO: 19, or (d) an amino acid sequence having 100% sequence identity to amino acids number 22 to 235 of sequence of SEQ ID NO: 19.

10. The method of claim 1, wherein: (i) population of modified immunoresponsive cells expressing or presenting a heterologous TCR further expresses or presents a heterologous co-stimulatory ligand; optionally 4-1BBL, or CD80; and/or (ii) the modified immunoresponsive cell is (a) a B cell, T cell or natural killer (NK) cell, (b) a T cell, optionally a CD4.sup.+ T cell or a CD8.sup.+ T cell.

11. The method of claim 1, wherein: (i) the PD-1 axis binding antagonist and modified immunoresponsive cells are administered separately, sequentially or simultaneously; (ii) the PD-1 axis binding antagonist is administered: (a) prior to, simultaneous with or after the modified immunoresponsive cells, (b) prior to and after the modified immunoresponsive cells, or (c) simultaneously with and after the modified immunoresponsive cells. (d) prior to and simultaneously with the modified immunoresponsive cells, (e) after the modified immunoresponsive cells, or (f) prior to and simultaneously with and after the modified immunoresponsive cells; and/or (iii) the PD-1 axis binding antagonist and/or the modified immunoresponsive cells are administered continuously or intermittently.

12. The method of claim 1, wherein: (i) the modified immunoresponsive cells are administered as a single dose; (ii) the modified immunoresponsive cells are administered at a dose of between about 500 million to about 1 billion cells, about 2 billion to about 5 billion cells or about 6 billion to about 10 billion cells; or (iii) the PD-1 axis binding antagonist is administered at a dose of: (a) between about 1 to 9 or about 10 to 20 mg/kg, (b) between about 3 to 5 mg/kg, (c) between about 50 to 200 mg or about 300 to 500 mg, or (d) about 200 mg, optionally wherein the dose is a fixed dose.

13. The method of claim 1 wherein: (i) the PD-1 axis binding antagonist is administered as (a) a single dose in each of one or more dosing cycles, (b) one or more doses in each of one or more dosing cycles, (c) a single dose on the first day of each of one or more dosing cycles, (d) one or more doses in each of one or more dosing cycles, at least one dose being on the first day of each cycle; (ii) the dosing cycle is (a) 14 to 17 days, 18 to 21 days, 22 to 24 days, 24 to 27 days, 28 to 30 days or 31 days, (b) one week, two weeks, three weeks, four weeks or one month; and/or (iii) the first dose of PD-1 axis binding antagonist after the modified immunoresponsive cells are administered is on either day 17, day 21 or day 22 after the immunoresponsive cells are administered.

14. The method of claim 1, wherein the PD-1 axis binding antagonist is administered as one or more doses on each of one or more dosing cycles, prior to administration of the modified immunoresponsive cells and is administered as one or more doses on each of one or more dosing cycles after administration of the modified immunoresponsive cells, optionally wherein: (i) (a) the PD-1 axis binding antagonist is administered as one or more doses on each of one or more dosing cycles prior to administration of the modified immunoresponsive cells, (b) the status of disease is determined in comparison to the status prior to PD-1 axis binding antagonist administration, wherein if stable disease or progressive disease is determined then, (c) modified immunoresponsive cells are administered and the PD-1 axis binding antagonist is administered as one or more doses on each of one or more dosing cycles after the administration of the modified immunoresponsive cells, optionally wherein the PD-1 axis binding antagonist is administered for a specified period, preferably 24 months; or (ii) (a) the PD-1 axis binding antagonist is administered as one or more doses on each of one or more dosing cycles prior to the administration of the modified immunoresponsive cells, (b) the status of disease is determined in comparison to the status prior to PD-1 axis binding antagonist administration, wherein if complete response or partial response is determined then, (c) the PD-1 axis binding antagonist is administered as one or more doses on each of one or more dosing cycles without administration of the modified immunoresponsive cells, optionally wherein, in step (c) stable disease or progressive disease is determined then, (d) modified immunoresponsive cells are administered and the PD-1 axis binding antagonist is administered as one or more doses on each of one or more dosing cycles after the administration of the modified immunoresponsive cells or simultaneously with and after the administration of the modified immunoresponsive cells, wherein in step (c) or (d) the PD-1 axis binding antagonist is administered for the shorter of a specified period or determination of disease progression, optionally wherein the specified period is 24 months.

15. The method of claim 1, wherein the PD-1 axis binding antagonist and/or modified immunoresponsive cells are administered intravenously or by intravenous infusion.

16. The method of claim 1, wherein: (i) the cancer is relapsed cancer or refractory cancer or recurrent cancer or locally recurrent cancer or metastatic cancer, non-resectable cancer or locally confined, cancer with no surgical or radiotherapy option or inoperable cancer; (ii) the subject has relapsed cancer or refractory cancer or recurrent cancer or has locally recurrent cancer or metastatic cancer or locally confined or inoperable cancer; (iii) the cancer is selected from; lung cancer, non-small cell lung cancer (NSCLC), metastatic or advanced NSCLC, squamous NSCLC, adenocarcinoma NSCLC, adenosquamous NSCLC, large cell NSCLC, ovarian cancer, gastric cancer, urothelial cancer, esophageal cancer, esophagogastric junction cancer (EGJ), melanoma, bladder cancer, head and neck cancer, head and neck squamous cell carcinoma (HNSCC), cancer of the oral cavity, cancer of the oropharynx, cancer of the hypopharynx, cancer of the throat, cancer of the larynx, cancer of the the tonsil, cancer of the tongue, cancer of the soft palate, cancer of the pharynx, synovial sarcoma, myxoid round cell liposarcoma (MRCLS), optionally wherein the cancer or tumour express a MAGE antigen or peptide antigen thereof and/or express PD-L1 or PD-L2 [optionally with CPS≥1], optionally MAGE-A4 antigen or peptide antigen thereof and/or express PD-L1 or PD-L2 [optionally with CPS≥1]; and/or (iv) the cancer is selected from any one of breast cancer, metastatic breast cancer, liver cancer, renal cell carcinoma, synovial sarcoma, urothelial cancer or tumour, pancreatic cancer, colorectal cancer, metastatic stomach cancer, metastatic gastric cancer, metastatic liver cancer, metastatic ovarian cancer, metastatic pancreatic cancer, metastatic colorectal cancer, metastatic lung cancer, colorectal carcinoma or adenocarcinoma, lung carcinoma or adenocarcinoma, pancreatic carcinoma or adenocarcinoma, mucinous adenoma, ductal carcinoma of the pancreas, hematological malignancy, optionally wherein the cancer or tumour express a MAGE antigen or peptide antigen thereof and/or express PD-L1 or PD-L2 [optionally with CPS≥1], optionally MAGE-A4 antigen or peptide antigen thereof and/or express PD-L1 or PD-L2 [optionally with CPS≥1].

17. The method of claim 1 wherein the subject has not received prior cancer treatment.

18. The method of claim 1, wherein the subject has received prior cancer treatment and/or has failed to respond to prior cancer treatment, optionally wherein: (a) the prior treatment comprises; systemic and/or local therapy, optionally any one or more of surgery, radiation therapy cryotherapy, laser therapy, topical therapy and/or systemic therapy, for example any one or more of chemotherapy, hormonal therapy, targeted drugs, or immunotherapy; (b) the prior treatment comprises a PD-1 axis binding antagonist, PD-L1 binding antagonist or PD-1 binding antagonist, optionally wherein the PD-1 axis binding antagonist is an antibody; (c) the prior treatment comprises an Epidermal Growth Factor Receptor Antagonist, optionally Cetuximab; (d) the prior treatment comprises chemotherapy comprising a platinum compound, optionally selected from Lipoplatin, Cisplatin, Carboplatin, Oxaliplatin, Nedaplatin, Triplatin tetranitrate, Phenanthriplatin, Satraplatin, Picoplatin: (e) the prior treatment comprises chemotherapy comprising a chemotherapeutic agent selected from, methotrexate, capecitabine, taxane, anthracycline, paclitaxel, docetaxel, paclitaxel protein bound particles, doxorubicine, epirubicine, 5-fluorouracil, cyclophosphamide, afatinib, vincristine, etoposide or combinations thereof; or (f) the prior treatment comprises chemotherapy comprising a chemotherapeutic agent selected from, FEC: 5-fluorouracil, epirubicine, cyclophosphamide; FAC: 5-fluorouracil, doxorubicine, cyclophosphamide; AC: doxorubicine, cyclophosphamide; EC: epirubicine, cyclophosphamide.

19. The method of claim 18, wherein: (i) the subject has not received prior treatment in recurrence less than or equal to 12 months since the last treatment or less than or equal to 6 months since the last treatment; or (ii) the subject has not received any prior adjuvant therapy (surgery followed by radiation and/or chemotherapy) in recurrence less than or equal to 12 months since the last treatment or in recurrence less than or equal to 6 months since the last treatment.

20. The method of claim 1, wherein the treatment effectively extends or improves: (a) the progression free survival, (b) the time to progression, (c) the duration of response, (d) the overall survival, (e) the objective response or objective response rate, (f) the overall response or overall response rate, (g) partial response or partial response rate, (h) complete response or complete response rate, (i) stable disease rate or median stable disease (j) median progression free survival, (k) median time to progression, (l) median duration of response, (m) median overall survival, (n) median objective response or median objective response rate, (o) median overall response or median overall response rate, (p) median partial response or median partial response rate, (q) median complete response or median complete response, (r) median stable disease rate or median stable disease, in comparison to treatment with PD-1 axis binding antagonist alone or modified immunoresponsive cells alone.

Description

FIGURES

[0237] FIG. 1. Increase in CD3 and PD-L1 detected in the post-infusion tumour of a responder

[0238] FIG. 2. increase in CD3 and PD-L1 post infusion in a non-responder

[0239] FIG. 3. MAGE-A4 SPEAR T-cells upregulate PD-1 expression upon stimulation in comparison to non-transduced (NTD) T-cells.

[0240] FIG. 4. MAGE-A4+CD8 SPEAR T-cells upregulate PD-1 expression upon stimulation in comparison to non-transduced (NTD) T-cells.

[0241] FIG. 5. PD-L1 is expressed and also upregulated by IFN-γ in A375 cells (A375.GFP MAGE-A4.sup.+ melanoma cell line)

[0242] FIG. 6. Scheme of pre-activation assay to stimulate and upregulate PD-1 expression on SPEAR T-cells (heterologous MAGE-A4 TCR or heterologous MAGE-A4 TCR+heterologous CD8 expressing T-cells).

[0243] FIG. 7. PD-1 is upregulated on A2M4 SPEAR T-cells by pre-activation.

[0244] FIG. 8. Supernatants of A2M4 cultures during (A) initial stimulation and (B) during re-stimulation with or without anti-PD-1 antibody. IFN-gamma ELISA data is shown here (n=6 small-scale donors).

[0245] FIG. 9. Scheme of combination A2M4 SPEAR T-cell+PD-1 axis binding antagonist (Pembrolizumab) treatment of cancer subject.

[0246]

TABLE-US-00001 Sequences PD1 - Human Programmed cell death protein (Homo sapiens) SEQ ID NO: 1 MQIPQAPWPVVWAVLQLGWRPGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSNTS ESFVLNWYRMSPSNQTDKLAAFPEDRSQPGQDCRFRVTQLPNGRDFHMSVVRARRNDSGT YLCGAISLAPKAQIKESLRAELRVTERRAEVPTAHPSPSPRPAGQFQTLVVGVVGGLLGS LVLLVWVLAVICSRAARGTIGARRTGQPLKEDPSAVPVFSVDYGELDFQWREKTPEPPVP CVPEQTEYATIVFPSGMGTSSPARRGSADGPRSAQPLRPEDGHCSWPL, PD1L1 - Human Programmed cell death 1 ligand 1(Homo sapiens) SEQ ID NO: 2 MRIFAVFIFMTYWHLLNAFTVTVPKDLYVVEYGSNMTIECKFPVEKQLDLAALIVYWEME DKNIIQFVHGEEDLKVQHSSYRQRARLLKDQLSLGNAALQITDVKLQDAGVYRCMISYGG ADYKRITVKVNAPYNKINQRILVVDPVTSEHELTCQAEGYPKAEVIWTSSDHQVLSGKTT TTNSKREEKLFNVTSTLRINTTTNEIFYCTFRRLDPEENHTAELVIPELPLAHPPNERTH LVILGAILLCLGVALTFIFRLRKGRMMDVKKCGIQDTNSKKQSDTHLEET, PD1L2 - Human Programmed cell death 1 ligand 2 (Homo sapiens) SEQ ID NO: 3 MIFLLLMLSLELQLHQIAALFTVTVPKELYIIEHGSNVTLECNFDTGSHVNLGAITASLQ KVENDTSPHRERATLLEEQLPLGKASFHIPQVQVRDEGQYQCIIIYGVAWDYKYLTLKVK ASYRKINTHILKVPETDEVELTCQATGYPLAEVSWPNVSVPANTSHSRTPEGLYQVTSVL RLKPPPGRNFSCVFWNTHVRELTLASIDLQSQMEPRTHPTWLLHIFIPFCIIAFIFIATV IALRKQLCQKLYSSKDTTKRPVTTTKREVNSAI, Durvalumab Heavy Chain Sequence SEQ ID NO: 4 EVQLVESGGGLVQPGGSLRLSCAASGFTFSRYWMSWVRQAPGKGLEWVANIKQDGSEKYY VDSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCAREGGWFGELAFDYWGQGTLVTVS SASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQS SGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPEFEG GPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQY NSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVYTLPPSRE EMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSR WQQGNVFSCSVMHEALHNHYTQKSLSLSPGK,  Durvalumab Light Chain Sequence SEQ ID NO: 5 EIVLTQSPGTLSLSPGERATLSCRASQRVSSSYLAWYQQKPGQAPRLLIYDASSRATGIP DRFSGSGSGTDFTLTISRLEPEDFAVYYCQQYGSLPWTFGQGTKVEIKRTVAAPSVFIFP PSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTL TLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC,  Atezolizumab Heavy Chain Sequence SEQ ID NO: 6 EVQLVESGGGLVQPGGSLRLSCAASGFTFSDSWIHWVRQAPGKGLEWVAWISPYGGSTYY ADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARRHWPGGFDYWGQGTLVTVSSAS TKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGL YSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPS VFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYAST YRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMT KNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQ GNVFSCSVMHEALHNHYTQKSLSLSPGK,  Atezolizumab Light Chain Sequence SEQ ID NO: 7 DIQMTQSPSSLSASVGDRVTITCRASQDVSTAVAWYQQKPGKAPKLLIYSASFLYSGVPS RFSGSGSGTDFTLTISSLQPEDFATYYCQQYLYHPATFGQGTKVEIKRTVAAPSVFIFPP SDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLT LSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC,  Avelumab Heavy Chain Sequence SEQ ID NO: 8 EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYIMMWVRQAPGKGLEWVSSIYPSGGITFY ADTVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARIKLGTVTTVDYWGQGTLVTVSS ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSS GLYSLSSVVTVPSSSLGTOTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGG PSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYN STYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDE LTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRW QQGNVFSCSVMHEALHNHYTQKSLSLSPGK, Avelumab Light Chain Sequence SEQ ID NO: 9 QSALTQPASVSGSPGQSITISCTGTSSDVGGYNYVSWYQQHPGKAPKLMIYDVSNRPSGV SNRFSGSKSGNTASLTISGLQAEDEADYYCSSYTSSSTRVFGTGTKVTVLGQPKANPTVT LFPPSSEELQANKATLVCLISDFYPGAVTVAWKADGSPVKAGVETTKPSKQSNNKYAASS YLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS, MDX1105 Heavy Chain Sequence SEQ ID NO: 10 QVQLVQSGAEVKKPGSSVKVSCKTSGDTFSTYAISWVRQAPGQGLEWMGGIIPIFGKAHYAQ KFQGRVTITADESTSTAY MELSSLRSEDTAVYFCARKFHFVSGSPFGMDVWGQGTTVTVSSASTKGPSVFPLAPSSKSTS GGTAALGCLVKDYFPEPV TVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVE PKSCDKT, MDX1105 Light Chain Sequence SEQ ID NO: 11 EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLIYDASNRATGIPARF SGSGSGTDFTLTISSLEP EDFAVYYCQQRSNWPTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPRE AKVQWKVDNALQSGNSQE SVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC,  Pembrolizumab - DB09037 >Heavy Chain Sequence SEQ ID NO: 12 QVQLVQSGVEVKKPGASVKVSCKASGYTFTNYYMYWVRQAPGQGLEWMGGINPSNGGTNF NEKFKNRVTLTTDSSTTTAYMELKSLQFDDTAVYYCARRDYRFDMGFDYWGQGTTVTVSS ASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSS GLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSV FLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTY RVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTK NQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEG NVFSCSVMHEALHNHYTQKSLSLSLGK, Pembrolizumab Light Chain Sequence SEQ ID NO: 13 EIVLTQSPATLSLSPGERATLSCRASKGVSTSGYSYLHWYQQKPGQAPRLLIYLASYLES GVPARFSGSGSGTDFTLTISSLEPEDFAVYYCQHSRDLPLTFGGGTKVEIKRTVAAPSVE IFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLS STLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC,  Nivolumab Heavy Chain Sequence SEQ ID NO: 14 QVQLVESGGGVVQPGRSLRLDCKASGITFSNSGMHWVRQAPGKGLEWVAVIWYDGSKRYY ADSVKGRFTISRDNSKNTLFLQMNSLRAEDTAVYYCATNDDYWGQGTLVTVSSASTKGPS VFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSS VVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKP KDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLT VLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTC LVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSV MHEALHNHYTQKSLSLSLGK, Nivolumab Light Chain Sequence SEQ ID NO: 15 EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLIYDASNRATGIPA RFSGSGSGTDFTLTISSLEPEDFAVYYCQQSSNWPRTFGQGTKVEIKRTVAAPSVFIFPP SDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLT LSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC, Cemiplimab Heavy Chain Sequence SEQ ID NO: 16 EVQLLESGGVLVQPGGSLRLSCAASGFTFSNFGMTWVRQAPGKGLEWVSGISGGGRDTYF ADSVKGRFTISRDNSKNTLYLQMNSLKGEDTAVYYCVKWGNIYFDYWGQGTLVTVSSAST KGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLY SLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLF PPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVV SVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQV SLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVF SCSVMHEALHNHYTQKSLSLSLGK, Cemiplimab Light Chain Sequence SEQ ID NO: 17 DIQMTQSPSSLSASVGDSITITCRASLSINTFLNWYQQKPGKAPNLLIYAASSLHGGVPS RFSGSGSGTDFTLTIRTLQPEDFATYYCQQSSNTPFTFGPGTVVDFRRTVAAPSVFIFPP SDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLT LSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC, MAGE A4 peptide SEQ ID NO: 18 GVYDGREHTV, (CD8α)CDRs bold underlined, signal sequence italic underlined SEQ ID NO: 19 MALPVTALLLPLALLLHAARPSQFRVSPLDRTWNLGETVELKCQ CSWLFQPRG AAASPTFLL AAEGLDTQRFSGKRLGDTFVLTLSDFRRENEGYYFCSA Y FSHFVPVFLPAKPTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACDIYIW APLAGTCGVLLLSLVITLYCNHRNRRRVCKCPRPVVKSGDKPSLSARYV; (CD8α) SEQ ID NO: 20 ATGGCCTTACCAGTGACCGCCTTGCTCCTGCCGCTGGCCTTGCTGCTCCACGCCGCCAGGCC GAGCCAGTTCCGGGTGTCGCCGCTGGATCGGACCTGGAACCTGGGCGAGACAGTGGAGCTGA AGTGCCAGGTGCTGCTGTCCAACCCGACGTCGGGCTGCTCGTGGCTCTTCCAGCCGCGCGGC GCCGCCGCCAGTCCCACCTTCCTCCTATACCTCTCCCAAAACAAGCCCAAGGCGGCCGAGGG GCTGGACACCCAGCGGTTCTCGGGCAAGAGGTTGGGGGACACCTTCGTCCTCACCCTGAGCG ACTTCCGCCGAGAGAACGAGGGCTACTATTTCTGCTCGGCCCTGAGCAACTCCATCATGTAC TTCAGCCACTTCGTGCCGGTCTTCCTGCCAGCGAAGCCCACCACGACGCCAGCGCCGCGACC ACCAACACCGGCGCCCACCATCGCGTCGCAGCCCCTGTCCCTGCGCCCAGAGGCGTGCCGGC CAGCGGCGGGGGGCGCAGTGCACACGAGGGGGCTGGACTTCGCCTGTGATATCTACATCTGG GCGCCCTTGGCCGGGACTTGTGGGGTCCTTCTCCTGTCACTGGTTATCACCCTTTACTGCAA CCACAGGAACCGAAGACGTGTTTGCAAATGTCCCCGGCCTGTGGTCAAATCGGGAGACAAGC CCAGCCTTTCGGCGAGATACGTCGGTTCAAGAGCTAAAAGAAGTGGTAGTGGTGCCCCTGTG A; (MAGE A4 TCR α chain) CDRs bold underlined SEQ ID NO: 21 MKKHLTTFLVILWLYFYRGNGKNQVEQSPQSLIILEGKNCTLQCNYT LRWYKQDTG RGPVSLTI TKSNGRYTATLDADTKQSSLHITASQLSDSASYI GAGTQVVVTPDIQNPDPAVYQLRDSKSSDKSVCLFTDFDSQTNVSQSKDSDVYITDKTVLD MRSMDFKSNSAVAWSNKSDFACANAFNNSIIPEDTFFPSPESSCDVKLVEKSFETDTNLNFQ NLSVIGFRILLLKVAGFNLLMTLRLWSSGSRAKR; (MAGE A4 TCR α chain coding sequence) SEQ ID NO: 22 ATGAAGAAGCACCTGACCACCTTTCTCGTGATCCTGTGGCTGTACTTCTACCGGGGCAACGG CAAGAACCAGGTGGAACAGAGCCCCCAGAGCCTGATCATCCTGGAAGGCAAGAACTGCACCC TGCAGTGCAACTACACCGTGTCCCCCTTCAGCAACCTGCGGTGGTACAAGCAGGACACCGGC AGAGGCCCTGTGTCCCTGACCATCCTGACCTTCAGCGAGAACACCAAGAGCAACGGCCGGTA CACCGCCACCCTGGACGCCGATACAAAGCAGAGCAGCCTGCACATCACCGCCAGCCAGCTGA GCGATAGCGCCAGCTACATCTGCGTGGTGTCCGGCGGCACAGACAGCTGGGGCAAGCTGCAG TTTGGCGCCGGAACACAGGTGGTCGTGACCCCCGACATCCAGAACCCTGACCCTGCCGTGTA CCAGCTGCGGGACAGCAAGAGCAGCGACAAGAGCGTGTGCCTGTTCACCGACTTCGACAGCC AGACCAACGTGTCCCAGAGCAAGGACAGCGACGTGTACATCACCGACAAGACCGTGCTGGAC ATGCGGAGCATGGACTTCAAGAGCAATAGCGCCGTGGCCTGGTCCAACAAGAGCGACTTCGC CTGCGCCAACGCCTTCAACAACAGCATTATCCCCGAGGACACATTCTTCCCAAGCCCCGAGA GCAGCTGCGACGTCAAGCTGGTGGAAAAGAGCTTCGAGACAGACACCAACCTGAACTTCCAG AACCTGAGCGTGATCGGCTTCAGAATCCTGCTGCTGAAGGTGGCCGGCTTCAACCTGCTGAT GACCCTGAGACTGTGGTCCAGCGGCAGCCGGGCCAAGAGA; (MAGE A4 TCR β chain) CDRs bold underlined SEQ ID NO: 23 MASLLFFCGAFYLLGTGSMDADVTQTPRNRITKTGKRIMLECSQT MYWYRQDPGLGL RLIYY INKGEISDGYSVSRQAQAKFSLSLESAIPNQTALYF G PGTRLTVLEDLKNVFPPEVAVFEPSEAEISHTQKATLVCLATGFYPDHVELSWWVNGKEVHS GVSTDPQPLKEQPALNDSRYCLSSRLRVSATFWQNPRNHFRCQVQFYGLSENDEWTQDRAKP VTQIVSAEAWGRADCGFTSESYQQGVLSATILYEILLGKATLYAVLVSALVLMAMVKRKDSR G; (MAGE A4 TCR β chain coding sequence) SEQ ID NO: 24 ATGGCCAGCCTGCTGTTCTTCTGCGGCGCCTTCTACCTGCTGGGCACCGGCTCTATGGATGC CGACGTGACCCAGACCCCCCGGAACAGAATCACCAAGACCGGCAAGCGGATCATGCTGGAAT GCTCCCAGACCAAGGGCCACGACCGGATGTACTGGTACAGACAGGACCCTGGCCTGGGCCTG CGGCTGATCTACTACAGCTTCGACGTGAAGGACATCAACAAGGGCGAGATCAGCGACGGCTA CAGCGTGTCCAGACAGGCTCAGGCCAAGTTCAGCCTGTCCCTGGAAAGCGCCATCCCCAACC AGACCGCCCTGTACTTTTGTGCCACAAGCGGCCAGGGCGCCTACGAGGAGCAGTTCTTTGGC CCTGGCACCCGGCTGACAGTGCTGGAAGATCTGAAGAACGTGTTCCCCCCAGAGGTGGCCGT GTTCGAGCCTTCTGAGGCCGAAATCAGCCACACCCAGAAAGCCACACTCGTGTGTCTGGCCA CCGGCTTCTACCCCGACCACGTGGAACTGTCTTGGTGGGTCAACGGCAAAGAGGTGCACAGC GGCGTGTCCACCGATCCCCAGCCTCTGAAAGAACAGCCCGCCCTGAACGACAGCCGGTACTG CCTGAGCAGCAGACTGAGAGTGTCCGCCACCTTCTGGCAGAACCCCAGAAACCACTTCAGAT GCCAGGTGCAGTTTTACGGCCTGAGCGAGAACGACGAGTGGACCCAGGACAGAGCCAAGCCC GTGACACAGATCGTGTCTGCCGAAGCTTGGGGGCGCGCCGATTGTGGCTTTACCAGCGAGAG CTACCAGCAGGGCGTGCTGAGCGCCACCATCCTGTACGAGATCCTGCTGGGAAAGGCCACAC TGTACGCCGTGCTGGTGTCTGCCCTGGTGCTGATGGCCATGGTCAAGCGGAAGGACAGCCGG GGC; (MAGE A4 TCR α chain variable region)136AA - CDRs bold underlined SEQ ID NO: 25 MKKHLTTFLVILWLYFYRGNGKNQVEQSPQSLIILEGKNCTLQCNYT LRWYKQDTG RGPVSLTI TKSNGRYTATLDADTKQSSLHITASQLSDSASYI GAGTQVVVTPD; (MAGE A4 TCR β chain variable region)133AA - CDRs bold underlined SEQ ID NO: 26 MASLLFFCGAFYLLGTGSMDADVTQTPRNRITKTGKRIMLECSQT MYWYRQDPGLGL RLIYY INKGEISDGYSVSRQAQAKFSLSLESAIPNQTALYF G PGTRLTVLE; CDR1 MAGE A4 TCR α chain, (residues 48-53) SEQ ID NO: 27 VSPFSN; CDR2 MAGE A4 TCR α chain, (residues 71-76) SEQ ID NO: 28 LTFSEN; CDR3 MAGE A4 TCR α chain, (residues 111-125) SEQ ID NO: 29 CVVSGGTDSWGKLQF; CDR1 MAGE A4 TCR β chain, (residues 46 - 50) SEQ ID NO: 30 KGHDR; CDR2 MAGE A4 TCR β chain, (residues 68-73) SEQ ID NO: 31 SFDVKD; CDR3 MAGE A4 TCR β chain, (residues 110 - 123) SEQ ID NO: 32 CATSGQGAYEEQFF; CDR1 CD8α (residues 45-53) SEQ ID NO: 33 VLLSNPTSG; CDR2 CD8α (residues 72-79) SEQ ID NO: 34 YLSQNKPK; CDR3 CD8α (residues 118-123) SEQ ID NO: 35 LSNSIM; MAGE A4 SEQ ID NO: 36 MSSEQKSQHC KPEEGVEAQE EALGLVGAQA PTTEEQEAAV SSSSPLVPGT LEEVPAAESA GPPQSPQGAS ALPTTISFTC WRQPNEGSSS QEEEGPSTSP DAESLFREAL SNKVDELAHF LLRKYRAKEL VTKAEMLERV IKNYKRCFPV IFGKASESLK MIFGIDVKEV DPASNTYTLV TCLGLSYDGL LGNNQIFPKT GLLIIVLGTI AMEGDSASEE EIWEELGVMG VYDGREHTVY GEPRKLLTQD WVQENYLEYR QVPGSNPARY EFLWGPRALA ETSYVKVLEH VVRVNARVRI AYPSLREAAL LEEEEGV,

EXAMPLES

Example 1

[0247] Using tissue biopsy samples taken before and after T-cell infusion (SPEAR T-cells i.e. comprising subject T-cells engineered with a heterologous TCR specific to MAGE A4 cancer testis antigen) from a subject synovial sarcoma patient (tumour expressing MAGE A4) we have shown that prior to the infusion there was no PD-L1 expression in the tissue. The patient was responsive to SPEAR T-cell treatment. Post SPEAR T-cell infusion, there was an increase in measured CD3 (part of the T cell receptor (TCR) complex on a mature T lymphocyte) expressed in the tissue signaling T-cell infiltration and the presence of the gene-modified T-cells. There was additionally an observed induction of PD-L1 in the tumor associated with the T-cell infiltration, FIG. 1. Data from biopsies from a subject ovarian cancer patient who was non-responsive to SPEAR T-cell treatment demonstrated that PD-L1 expression was positive in the pre-infusion biopsy and that following SPEAR T-cell infusion there was an increase in T-cell/gene-modified T-cell infiltration (indicated by CD3) and an observed associated stronger induction of PD-L1 in the tumor, FIG. 2.

[0248] We have shown that exposure of gene-modified T-cells expressing heterologous TCR to MAGE-A4 upregulate PD-1 expression upon stimulation with the target antigen MAGE-A4 in comparison to non-transduced (NTD) T-cells not having the heterologous TCR, FIG. 3. The same effect is seen in gene-modified T-cells expressing heterologous MAGE-A4 and heterologous CD8, FIG. 4. The data indicates that the activity of the engineered T-cells could be enhanced by blocking the PD1/PD-L1 interaction. We have also shown that cancer cell lines upregulate PD-L1 in response to interferon, FIG. 5. The data of FIG. 5 demonstrates that what is seen clinically in patients is also seen in the in-vitro cancer cell assay system. As part of the immune response to cancer T-cells produce interferon which has an anti-tumour effect through the JAK/STAT pathway of the target cell. In patients the PD-L1 upregulation response of tumour to interferon represents part of the immune evasion adaptations of tumour cells.

[0249] We have further examined the effect of PD-1 blockade on the cytotoxicity and effector cytokine production by gene-modified T-cells expressing heterologous TCR to MAGE-A4 (SPEAR T cells) using a pre-stimulation protocol and ELISA assay for cytokines. T-cell pre-activation was performed as set out in FIG. 6, in which gene-modified T-cells expressing heterologous TCR to MAGE-A4 (also performed for gene-modified T-cells expressing both heterologous MAGE-A4 and heterologous CD8) are stimulated with target expressing tumour cells (irradiated A375 MAGE-A4.sup.+ melanoma cells), cultured and separated from the cancer cells prior to re-stimulation in the presence of absence of PD-1 blockade with anti-PD-1 antibody. The data of FIG. 7 shows that the expression of PD-1 on the pre-activated T-cells over the period of the initial stimulation of 7 days increases for both CD4+ and CD8+ cells over each population.

[0250] Using the pre-stimulation model we have shown that IFN-gamma, IL-2 and granzyme B are produced during the initial stimulation in both engineered MAGE-A4 TCR and engineered MAGE-A4 TCR+CD8 T-cells during the initial stimulation but this ability is lost after this pre-activation step on re-stimulation. However, re-stimulation in the presence of anti-PD-1 antibody partially recovers this cytokine activity function and restores IFN gamma and granzyme B but not IL-2 production for both engineered MAGE-A4 TCR and engineered MAGE-A4 TCR+CD8 T-cells, FIG. 8 shows data for MAGE-A4 TCR and IFN-gamma. In conclusion we have shown the ability to rescue exhausted engineered T-cell cytokine function by combination with anti-PD-1 antibody.

Example 2, In Vivo Tumour Model

[0251] We have further tested the combination in a NSG strain mouse A375 (melanoma) xenograft model using a dosing of 1×10.sup.6 transduced (ADP-A2M4) MAGE-A4 TCR T cells; 10 mg/kg intra peritoneal q4d (twice a day for four days) anti-PD-1 antibody pembrolizumab (Keytruda) and the effect on tumour volume recorded according to the following protocol. 72 tumour bearing mice are randomised into 9 groups of n=8, and treated as per Table 1 below.

TABLE-US-00002 TABLE 1 Treatment 1 Treatment 2 Scheduled Group Dosing Dosing euthanasia 01 Isotype control (IgG4) — Day 21 (n = 8) Q4D from Day 0, i.p. 02 Pembrolizumab — Day 21 (n = 8) Q4D from Day 0, i.p. 10 mg/kg 03 NTD T-cells — Day 21 (n = 8) Day 0 only, i.v. (1 × 10{circumflex over ( )}6cells) 04 NTD T-cells Pembrolizumab Day 21 (n = 8) Day 0 only, i.v. Q4D from Day 1, i.p. (1 × 10{circumflex over ( )}6cells) 10 mg/kg 05 T-cells Isotype control (IgG4) Day 60 (n = 8) Day 0 only, i.v. Q4D from Day 1, i.p. (1 × 10{circumflex over ( )}6cells) 06 T-cells Pembrolizumab Day 60 (n = 8) Day 0 only, i.v. Q4D from Day −1, i.p. (1 × 10{circumflex over ( )}6cells) 10 mg/kg 07 T-cells Pembrolizumab Day 60 (n = 8) Day 0 only, i.v. Q4D from Day 7, i.p. (1 × 10{circumflex over ( )}6cells) 10 mg/kg 08 T-cells Pembrolizumab Day 60 (n = 8) Day 0 only, i.v. Q4D from Day 14, i.p. (1 × 10{circumflex over ( )}6cells) 10 mg/kg 09 T-cells Pembrolizumab Day 60 (n=8) Day 0 only, i.v. Q4D from Day 21, i.p. (1 × 10{circumflex over ( )}6cells) 10 mg/kg

[0252] All animals in groups 1 and 2 will be administered either isotype control (group 1) or pembrolizumab (group 2) Q4D from Day 0 (i.e. Day 0, 4, 8, 12, 16, 20) via i.p. (maximum dosing volume via i.p. 10 ml/kg). No other treatment will be administered. (Day 0 is the day on which T-cells are administered as per groups 3 to 9).

[0253] All animals in groups 3 and 4 will receive a single administration of NTD T-cells on Day 0 via i.v. at a recommended volume of 5 ml/kg. No other treatment will be administered to animals in group 3. In addition, animals in group 4 will receive pembrolizumab Q4D from Day 1 (i.e. Day 1, 5, 9, 13, 17) via i.p. (maximum dosing volume via i.p. 10 ml/kg)

[0254] All animals in groups 5, 6, 7, 8 and 9 will receive a single administration of T-cells on Day 0 via i.v. at a recommended volume of 5 ml/kg and groups 5, 6, 7, 8 and 9 a second treatment as described below: [0255] Group 5: isotype control Q4D from Day 1 (i.e. Day 1, 5, 9, 13, 17, 21, 25, 29, 33, 37, 41, 45, 49, 53, 57) via i.p. (maximum dosing volume via i.p. 10 ml/kg) [0256] Group 6: pembrolizumab Q4D from Day −1 (i.e. Day −1, 3, 7, 11, 15, 19, 23, 27, 31, 35, 39, 43, 47, 51, 55, 59) via i.p. (maximum dosing volume via i.p. 10 ml/kg) [0257] Group 7: pembrolizumab Q4D from Day 7 (i.e. Day 7, 11, 15, 19, 23, 27, 31, 35, 39, 43, 47, 51, 55, 59) via i.p. (maximum dosing volume via i.p. 10 ml/kg) [0258] Group 8: pembrolizumab Q4D from Day 14 (i.e. Day 14, 18, 22, 26, 30, 34, 38, 42, 46, 50, 54, 58) via i.p. (maximum dosing volume via i.p. 10 ml/kg) [0259] Group 9: pembrolizumab Q4D from Day 21 (i.e. Day 21, 25, 29, 33, 37, 41, 45, 49, 53, 57) via i.p. (maximum dosing volume via i.p. 10 ml/kg)

[0260] In all cases tumour volumes are measured with callipers 3 times per week and animals weighed 3 times per week from initiation of treatment.

Example 3, Clinical Investigation

[0261] An investigation was designed to investigate combination treatment with Pembrolizumab with ADP-A2M4 (SPEAR T-cell, T-cell engineered to express MAGE A4 specific TCR) for the treatment of patients with recurrent or metastatic HNSCC in which the subject has received no prior systemic therapy for metastatic disease or subject has disease progression on or after platinum containing chemotherapy. Disease may be histologically or cytogenetically confirmed and/or measurable disease according to RECIST v1.1.

[0262] The study is a single arm Phase II study (n=10 patients) in advanced/recurrent Head and neck squamous cell carcinoma (HNSCC) with checkpoint inhibitor naïve patients in which the subject has received 0-1 prior lines of platinum containing systemic therapy for metastatic disease or in which the subject is checkpoint inhibitor naïve at screening prior to treatment and is due to start pembrolizumab treatment, or has recently started pembrolizumab treatment for advanced disease (having received 1-2 doses). Subjects are determined as PD-L1+, HLA-A2+, MAGE-A4 expression: 2+/3+ in ≥30% cells by IHC [i.e. tumour shows MAGE-A4 expression defined as ≥30% of tumour cells that are ≥2+ by IHC (ImmunoHistoChemistry)]. The primary endpoint for measure is ORR by RECIST v1.1. Historic response rates of standard of care PD-1 therapy, pembrolizumab monotherapy, in advanced/recurrent Head and neck squamous cell carcinoma (HNSCC) produces an ORR=19%.

[0263] Prior to initiating pembrolizumab treatment the subject is apheresed to obtain the T-cell population, the cells are subsequently transduced with the ADP-A2M4 “SPEAR” TCR specific for MAGE A4 antigen (particularly the specific MAGE A4 antigenic peptide SEQ ID NO:18) and the cells expanded and cryopreserved for later use. The subject is thereafter treated with up to 3 doses of pembrolizumab (200 mg IV infusion over 30 mins every 3 weeks) and then scanned for disease status, for example measurable disease according to RECIST v1.1. Subjects with progressive disease or stable disease go forward for a lymphodepletion regimen, fludarabine (30 mg/m.sup.2/day for 4 days) and cyclophosphamide (600 mg/m.sup.2/day for 3 days) and SPEAR T cell infusion: ADP-A2M4 dosing of 1 billion to 10 billion transduced cells with no dose escalation and continue pembrolizumab (200 mg IV infusion over 30 mins every 3 weeks) until disease progression or until 24 months whichever is the earlier. Subjects scanned and determined to have complete or partial response do not undergo lymphodepletion and cell infusion but continue pembrolizumab (200 mg IV infusion over 30 mins every 3 weeks) until disease progression or until 24 months whichever is the earlier, after which they may then undergo lymphodepletion and cell infusion and combination pembrolizumab administration. Optionally apheresis is to occur prior to start of pembrolizumab. First dose of pembrolizumab following cell infusion may be on day 17 or 22 following the cell infusion.

[0264] For the present study the subject exclusion criteria include: (a) HLA-A*02:05 positive in either allele. (b) HLA-A*02:07 (and alleles having the same protein sequence in the antigen binding domains as A*02:07) or any A*02 null allele (designated with an “N” suffix, e.g. A*02:32N) as the sole HLA-A*02 allele (e.g. a subject with HLA alleles A*02:04 and A*02:07 is eligible), (c) CNS metastases, (d) any prior checkpoint inhibitor therapy, (e) any prior cell therapy, (f) if received chemotherapy, wash out period required before leukapheresis or LD is 3 weeks.

[0265] The primary endpoint for measure is ORR by RECIST v1.1. The treatment scheme is presented in FIG. 9.