High avidity HPV T-cell receptors

Abstract

The present invention pertains to novel high avidity antigen recognizing constructs against Human Papilloma Virus antigens. The invention provides novel T cell receptor (TCR) based molecules which are selective and specific for HPV 16/18 proteins E5, E6 and E7. The TCR of the invention, and HPV antigen-binding fragments derived therefrom, are of use for the diagnosis, treatment and prevention of HPV infection, as well as for the diagnosis, treatment and prevention of HPV infection mediated secondary diseases as HPV infection caused cancers, such as cervical, nasopharyngeal or head and neck cancer. Further provided are nucleic acids encoding the proteins of the invention, and recombinant cells expressing the same.

Claims

1. An antigen recognizing construct comprising a T cell receptor (TCR), or an antigen binding fragment thereof, wherein the TCR or the antigen binding fragment thereof is composed of a TCR α chain sequence and a TCR β chain sequence, wherein (i) the TCR α chain sequence comprises an α chain complementary determining region 3 (CDR3), wherein an amino acid sequence of the α chain CDR3 comprises only one sequence selected from the group consisting of SEQ ID Nos: 9, 10, 12, 14, 16, 18, 21, 23, 25, and 27, and (ii) the TCR β chain sequence comprises a β chain CDR3; wherein an amino acid sequence of the β chain CDR3 comprises only one sequence selected from the group consisting of SEQ ID Nos: 11, 13, 15, 17, 19, 20, 22, 24, 26, and 28.

2. The antigen recognizing construct according to claim 1, wherein said TCR comprises a CDR1 and a CDR2 having sequences as set forth in SEQ ID NOs: 29 and 31; or 30 and 31; or 32 and 33; or 34 and 35; or 36 and 37; or 38 and 39; or 38 and 40; or 41 and 42; or 43 and 44; or 45 and 46; or 47 and 48.

3. The antigen recognizing construct according to claim 1, wherein said TCR comprises variable sequences as set forth in SEQ ID NOs: 29 and 31; or 30 and 31; or 32 and 33; or 34 and 35; or 36 and 37; or 38 and 39; or 38 and 40; or 41 and 42; or 43 and 44; or 45 and 46; or 47 and 48.

4. A nucleic acid encoding for an antigen recognizing construct according to claim 1.

5. A vector comprising a nucleic acid according to claim 4.

6. An isolated host cell comprising an antigen recognizing construct according to claim 1.

7. The host cell according to claim 6, comprising a lymphocyte.

8. An isolated host cell comprising a nucleic acid according to claim 4.

9. An isolated host cell comprising a vector according to claim 5.

10. The antigen recognizing construct according to claim 1, wherein the amino acid sequence of the α chain CDR3 consists of SEQ ID No. 9 or 10, and wherein the amino acid sequence of the β chain CDR consists of SEQ ID No. 11.

11. The antigen recognizing construct according to claim 1, wherein the amino acid sequence of the α chain CDR3 consists of SEQ ID No. 12, and wherein the amino acid sequence of the β chain CDR consists of SEQ ID No. 13.

12. The antigen recognizing construct according to claim 1, wherein the amino acid sequence of the α chain CDR3 consists of SEQ ID No. 14, and wherein the amino acid sequence of the β chain CDR consists of SEQ ID No. 15.

13. The antigen recognizing construct according to claim 1, wherein the amino acid sequence of the α chain CDR3 consists of SEQ ID No. 16, and wherein the amino acid sequence of the β chain CDR consists of SEQ ID No. 17.

14. The antigen recognizing construct according to claim 1, wherein the amino acid sequence of the α chain CDR3 consists of SEQ ID No. 18, and wherein the amino acid sequence of the β chain CDR consists of SEQ ID No. 20.

15. The antigen recognizing construct according to claim 1, wherein the amino acid sequence of the α chain CDR3 consists of SEQ ID No. 21, and wherein the amino acid sequence of the β chain CDR consists of SEQ ID No. 22.

16. The antigen recognizing construct according to claim 1, wherein the amino acid sequence of the α chain CDR3 consists of SEQ ID No. 23, and wherein the amino acid sequence of the β chain CDR consists of SEQ ID No. 24.

17. The antigen recognizing construct according to claim 1, wherein the amino acid sequence of the α chain CDR3 consists of SEQ ID No. 25, and wherein the amino acid sequence of the β chain CDR consists of SEQ ID No. 26.

18. The antigen recognizing construct according to claim 1, wherein the amino acid sequence of the α chain CDR3 consists of SEQ ID No. 27, and wherein the amino acid sequence of the β chain CDR consists of SEQ ID No. 28.

19. The antigen recognizing construct according to claim 1, wherein the amino acid sequence of the α chain CDR3 consists of SEQ ID No, 19, and wherein the amino acid sequence of the β chain CDR consists of SEQ ID No. 20.

Description

(1) The present invention will now be further described in the following examples with reference to the accompanying figures and sequences, nevertheless, without being limited thereto. For the purposes of the present invention, all references as cited herein are incorporated by reference in their entireties. In the Figures and Sequences:

(2) FIG. 1: shows examples for the specific CD8+ T cell response against HLA-A*02.01 restricted HPV16 E6 epitope TIHDIILECV in ABabDII mice. ABabDII mice were either immunized i.p. with E6/7 expressing adenovirus followed by s.c. administration of 50 μg of E6 peptide together with 50 μg of CpG ODN 1826 emulsified in IFA, or with peptide/CpG/IFA mixture alone. The presence of HPV-specific CD8+ T cells in the peripheral blood of immunized animals was assessed by intracellular IFNγ staining 7 days after each boost. Stimulation with phorbol myristate acetate (PMA) and ionomycin was used as a positive control (+ctrl).

(3) FIG. 2: shows an example for the specific CD8+ T cell response against HLA-A201 restricted HPV16 E7 epitope TLGIVCPI in ABabDII mice. ABabDII mice were immunized s.c. at the tail base with 50 μg of E7 peptide together with 50 μg of CpG ODN 1826 emulsified in IFA. The presence of HPV-specific CD8+ T cells in the peripheral blood of immunized animals was assessed by intracellular IFNγ staining 7 days after each boost. Stimulation with phorbol myristate acetate (PMA) and ionomycin was used as a positive control (+ctrl).

(4) FIG. 3: shows FACSort strategy (IFNγ capture assay) for the specific CD8+ T cells against HLA-A*02.01 restricted HPV16 E5, E6 and E7 epitopes. Spleen cells from responsive mice were depleted for CD4+ T cells and restimulated for 10 days with 10.sup.−8 mol of the respective peptide. For 5′RACE PCR HPV-specific CD8+ T cells were sorted directly into RNA purification buffer using IFNγ capture assay.

(5) FIG. 4: shows HPV16 E6 and E7 TCR transduction of freshly prepared human PBLs. 1×10.sup.6 freshly isolated or frozen hPBMCs were stimulated with anti-CD3 and anti-CD28-coated plates in the presence of 200 U/ml recombinant human interleukin 2. Transductions were done 48 and 72 h after stimulation by addition of retrovirus containing supernatant and protamine sulfate followed by spinoculation (1.sup.st transduction) or preloading of virus onto retronectin (Takara)-coated plates and spinoculation (2.sup.nd transduction). Transduction efficacy was assessed by staining with anti-CD8 and anti-mouse constant TCRβ mAbs.

(6) FIG. 5: shows the HPV16 E6/E7 peptide-specific IFN-γ release of TCR transduced human PBMCs; 5859 TCR corresponds to Seq ID No: 34 & 35, 5843B14 TCR corresponds to Seq ID No: 36 & 37, 14800 TCR corresponds to Seq ID NO: 43&44. IFNγ production was measured by enzyme-linked immunosorbent assay after 16 h coculture of 1×10.sup.4 TCR-transduced T cells with 1×10.sup.4 peptide-loaded T2 cells. Stimulation with phorbol myristate acetate (PMA) and ionomycin was used as a positive control (max); ut=untransduced PBMCs.

(7) FIG. 6: shows the HPV16 E6 IFN-γ release of TCR transduced human PBMCs on titrated E6-specific peptide and CASKI cell line. Left panel shows the TCR of SEQ ID NO 14/15 (T3), right panel shows TCR of SEQ ID NO 16/17 (T4). IFNγ production was measured by enzyme-linked immunosorbent assay after 16 h coculture of 1×10.sup.4 TCR-transduced T cells with 1×10.sup.4 peptide-loaded T2 cells or 1×10.sup.4 cells of CASKI cell line. Stimulation with phorbol myristate acetate (PMA) and ionomycin was used as a positive control (P+I). IFNγ release comparable to recognition of CASKI cells was also seen for HLA-transduced cervical cancer cell lines SiHa cell line and head and neck cancer cell lines SCC-090 and SCC-152 (not shown).

(8) FIG. 7: shows the HPV16 E5 transduction of freshly prepared human PBLs and HPV16 E5 peptide-specific IFN-γ release of TCR transduced human PBMCs. TCR used was the TCR of SEQ ID NO 12/13 (T2). 1×10.sup.6 freshly isolated or frozen hPBMCs were stimulated with anti-CD3 and anti-CD28-coated plates in the presence of 200 U/ml recombinant human interleukin 2. Transductions were done 48 and 72 h after stimulation by addition of retrovirus containing supernatant and protamine sulfate followed by spinoculation (1.sup.st transduction) or preloading of virus onto retronectin (Takara)-coated plates and spinoculation (2.sup.nd transduction). Transduction efficacy was assessed by staining with anti-CD8 and anti-mouse constant TCRβ mAbs. IFNγ production was measured by enzyme-linked immunosorbent assay after 16 h coculture of 1×10.sup.4 TCR-transduced T cells with 1×10.sup.4 peptide-loaded T2 cells. Stimulation with phorbol myristate acetate (PMA) and ionomycin was used as a positive control (P+I).

(9) SEQ ID No 1 to 8: show the HPV 16 and 18 epitopes that are bound by the TCR of the invention.

(10) SEQ ID No 9 to 28: show alpha and beta chain CDR3 sequences of the TCR of the invention.

(11) SEQ ID No 29 to 48: show the full variable sequences of α and β chains of the TCR of the present invention.

EXAMPLES

(12) HPV Epitopes used for Immunization of Animals:

(13) TABLE-US-00001 TABLE 1 HPV SEQ Gene Protein HLA A2.01 Epitope ID NO: 16-E5 Human papillomavirus type 16 E5 protein YIIFVYIPL (63-71) 1 16-E6 Human papillomavirus type 16 E6 protein KLPQLCTEL (11-19) 2 16-E6 Human papillomavirus type 16 E6 protein TIHDIILECV (29-38) 3 16-E7 Human papillomavirus type 16 E7 protein YMLDLQPET (86-93) 4 16-E7 Human papillomavirus type 16 E7 protein YMLDLQPETT (11-20) 5 16-E7 Human papillomavirus type 16 E7 protein TLGIVCPI (86-93) 6 18-E6 Human papillomavirus type 18 E6 protein KCIDFYSRI (67-75) 7 18-E7 Human papillomavirus type 18 E7 protein FQQLFLNTL (86-94) 8

(14) Peptide Epitopes with CTL reactivity in ABabDII mice

Example 1: T-Cell Receptor T1 Recognizing HPV 16-E5 Epitope YIIFVYIPL

(15) TABLE-US-00002 Immunogenic CDR3 peptide/ SEQ TCR Antigen purification TCR sequence ID NO: T1* E5 YIIFVYIPL TRAV13-1*01-CAASSYNQGGKLF-TRAJ23*-1  9 IFNγ- TRAV26-2*01-CILRDVNAGGTSYGKLTF-TRAJ52*01 10 CAPTURE TRBV12-3*01-CASSLLSSYNEQFF-TRBD2*01-TRBJ2-3*01 11

(16) CDR1 sequences are bold

(17) CDR2 sequences are double underlined

(18) CDR3 sequences are underlined

(19) Full length variable chain sequences (CDR3 sequences are underlined):

(20) TABLE-US-00003 TRAV13-1*01-CAASSYNQGGKLIF-TRAJ23*01 (SEQ ID NO: 29) MTSIRAVF IFLWLQLDLV NGENVEQHPS TLSVQEGDSA VIKCTYSDSA SNYFPWYKQE LGKGPQLIID IRSNVGEKKD QRIAVTLNKT AKHFSLHITE TQPEDSAVYF CAASSYNQGG KLIFGQGTEL SVKPN TRAV26-2*01-CILRDVNAGGTSYGKLTF-TRAJ52*01 (SEQ ID NO: 30) MKLVTSIT VLLSLGIMGD AKTTQPNSME SNEEEPVHLP CNHSTISGTD YIHWYRQLPS QGPEYVIHGL TSNVNNRMAS LAIAEDRKSS TLILHRATLR DAAVYYCILR DVNAGGTSYG KLTFGQGTIL TVHPN TRBV12-3*01-CASSLLSSYNEQFF-TRBD2*01-TRBJ2-3*01 (SEQ ID NO: 31) MDSWTF CCVSLCILVA KHTDAGVIQS PRHEVTEMGQ EVTLRCKPIS GHNSLFWYRQ TMMRGLELLI YFNNNVPIDD SGMPEDRFSA KMPNASFSTL KIQPSEPRDS AVYFCASSLL SSYNEQFFGP GTRLTVL

Example 2: T-Cell Receptor T2 Recognizing HPV 16-E5 Epitope YIIFVYIPL

(21) TABLE-US-00004 Immunogenic CDR3 peptide/ SEQ TCR Antigen purification TCR sequence ID NO: T2 E5 YIIFVYIPL TRAV12-2*02-CAVNVDFNKFYF-TRAJ21*01 12 IFNγ- TRBV4-1*01-CASSQDWNNEQFF-TRBD1*01-TRBJ2-1*01 13 CAPTURE

(22) Full length variable chain sequences (CDR3 sequences are underlined):

(23) TABLE-US-00005 TRAV12-2*02-CAVNVDFNKFYF-TRAJ21*01 (SEQ ID NO): 32: MMKSLRVLLV ILWLQLSWVW SQQKEVEQNS GPLSVPEGAI ASLNCTYSDR GSQSFFWYRQ YSGKSPELIM SIYSNGDKED GRFTAQLNKA SQYVSLLIRD SQPSDSATYL CAVNVDFNKF YFGSGTKLNV KPN TRBV4-1*01-CASSQDWNNEQFF-TRBD1*01-TRBJ2-1*01 SEQ ID NO: 33: MGCRLL CCAVLCLLGA VPIDTEVTQT PKHLVMGMTN KKSLKCEQHM GHRAMYWYKQ KAKKPPELMF VYSYEKLSIN ESVPSRFSPE CPNSSLLNLH LHALQPEDSA LYLCASSQDW NNEQFFGPGT RLTVL

Example 3: T-Cell Receptor T3 Recognizing HPV 16-E6 Epitope TIHDIILECV

(24) TABLE-US-00006 Immunogenic CDR3 peptide/ SEQ TCR Antigen purification TCR sequence ID NO: T3 E6 TIHDIILECV TRAV20*02-CA 14 VQANRGSTLGRL YF-TRAJ18*01 IFNγ- TRBV28*01-CA 15 CAPTURE SSLWGRLAKNIQ YF-TRBD1*01- TRBJ2-4*01

(25) Full length variable chain sequences (CDR3 sequences are underlined):

(26) TABLE-US-00007 TRAV20*02-CAVQANRGSTLGRLYF-TRAJ18*01 SEQ ID NO: 34 MEKMLEC AFIVLWLQLG WLSGEDQVTQ SPEALRLQEG ESSSLNCSYT VSGLRGLFWY RQDPGKGPEF LFTLYSAGEE KEKERLKATL TKKESFLHIT APKPEDSATY LCAVQANRGS TLGRLYFGRG TQLTVWPD TRBV28*01-CASSLWGRLAKNIQYF-TRBD1*01-TRBJ2-4*01 SEQ ID NO: 35 MGIRLL CRVAFCFLAV GLVDVKVTQS SRYLVKRTGE KVFLECVQDM DHENMFWYRQ DPGLGLRLIY FSYDVKMKEK GDIPEGYSVS REKKERFSLI LESASTNQTS MYLCASSLWG RLAKNIQYFG AGTRLSVL

Example 4: T-Cell Receptor T4 Recognizing HPV 16-E6 Epitope TIHDIILECV

(27) TABLE-US-00008 Immunogenic CDR3 peptide/ SEQ TCR Antigen purification TCR sequence ID NO: T4 E6 TIHDIILECV TRAV21*02-CA 16 VRETSGSRLTF- TRAJ58*01 IFNγ- TRBV28*01-CA 17 CAPTURE SSFWGRSTDTQY F-TRBD1*01- TRBJ2-3*01

(28) Full length variable chain sequences (CDR3 sequences are underlined):

(29) TABLE-US-00009 TRAV21*02-CAVRETSGSRLTF-TRAJ58*01 SEQ ID NO: 36 METLLGLL ILWLQLQWVS SKQEVTQIPA ALSVPEGENL VLNCSFTDSA IYNLQWFRQD PGKGLTSLLL IQSSQREQTS GRLNASLDKS SGRSTLYIAA SQPGDSATYL CAVRETSGSR LTFGEGTQLT VNPD TRBV28*01-CASSFWGRSTDTQYF-TRBD1*01-TRBJ2-3*01 SEQ ID NO: 37  MGIRLL CRVAFCFLAV GLVDVKVTQS SRYLVKRTGE KVFLECVQDM DHENMFWYRQ DPGLGLRLIY FSYDVKMKEK GDIPEGYSVS REKKERFSLI LESASTNQTS MYLCASSFWG RSTDTQYFGP GTRLTVL

Example 5: T-Cell Receptor T5 Recognizing HPV 16-E6 Epitope TIHDIILECV

(30) TABLE-US-00010 Immunogenic CDR3 peptide/ SEQ TCR Antigen purification TCR sequence ID NO: T5* E6 TIHDIILECV TRAV17*01-CA 18 TVSTDSWGKKLQ F-TRAJ24*02 IFNγ- TRBV10-3*02- 19 CAPTURE CAISDSNGINIQ YF-TRBD2*02- TRBJ2-4*01 TRBV28*01-CA 20 SSLWGRAGKDTQ YF-TRBD2*02- TRBJ2-3*01

(31) Full length variable chain sequences (CDR3 sequences are underlined):

(32) TABLE-US-00011 TRAV17*01-CATVSTDSWGKKLQF-TRAJ24*02 SEQ ID NO: 38 M ETLLGVSLVI LWLQLARVNS QQGEEDPQAL  SIQEGENATM NCSYKTSINN LQWYRQNSGR GLVHLILIRS NEREKHSGRL RVTLDTSKKS SSLLITASRA ADTASYFCAT VSTDSWGKLQ FGAGTQVVVT PD TRBV10-3*02-CAISDSNGINIQYF-TRBD2*02-TRBJ2-4*01 SEQ ID NO: 39 MRSWPG PEMGTRLFFY VALCLLWTGH MDAGITQSPR HKVTETGTPV TLRCHQTENH RYMYWYRQDP GHGLRLIHYS YGVKDTDKGE VSDGYSVSRS KTEDFLLTLE SATSSQTSVY FCAISDSNGI NIQYFGAGTR LSVL TRBV28*01-CASSLWGRAGKDTQYF-TRBD2*02-TRBJ2-3*01 SEQ ID NO: 40 MGIRLL CRVAFCFLAV GLVDVKVTQS SRYLVKRTGE KVFLECVQDM DHENMFWYRQ DPGLGLRLIY FSYDVKMKEK GDIPEGYSVS REKKERFSLI LESASTNQTS MYLCASSLWG RAGKDTQYFG PGTRLTVL

Example 6: T-Cell Receptor T6 Recognizing HPV 16-E6 Epitope KLPQLCTEL

(33) TABLE-US-00012 Immunogenic CDR3 peptide/ SEQ TCR Antigen purification TCR sequence ID NO: T6 E6 KLPQLCTEL TRAV8-6*01-C 21 AVSLNSGNTPLV F-TRAJ29*01 IFNγ- TRBV20-1*01- 22 CAPTURE CSARDLAGNTGE LFF-TRBD2*01- TRBJ2-2*01

(34) Full length variable chain sequences (CDR3 sequences are underlined):

(35) TABLE-US-00013 TRAV8-6*01-CAVSLNSGNTPLVF-TRAJ29*01 SEQ ID NO: 41 ML LLLVPAFQVI FTLGGTRAQS VTQLDSQVPV  FEEAPVELRC NYSSSVSVYL FWYVQYPNQG LQLLLKYLSG STLVESINGF EAEFNKSQTS FHLRKPSVHI SDTAEYFCAV SLNSGNTPLV FGKGTRLSVI AN TRBV20-1*01-CSARDLAGNTGELFF-TRBD2*01-TRBJ2-2*01 SEQ ID NO: 42 MLLLLLLLGP GSGLGAVVSQ HPSWVICKSG TSVKIECRSL DFQATTMFWY RQFPKQSLML MATSNEGSKA TYEQGVEKDK FLINHASLTL STLTVTSAHP EDSSFYICSA RDLAGNTGEL FFGEGSRLTVL

Example 7: T-Cell Receptor T7 Recognizing HPV 16-E7 Epitope TLGIVCPI

(36) TABLE-US-00014 Immunogenic CDR3 peptide/ SEQ TCR Antigen purification TCR sequence ID NO: T7 E7 TLGIVCPI TRAV30*01-CG 23 TGTDSWGKLQF- TRAJ24*02 IFNγ- TRBV12-4*01- 24 CAPTURE CASSPGLAGGEQ FF-TRBD2*02- TRBJ2-1*01

(37) Full length variable chain sequences (CDR3 sequences are underlined):

(38) TABLE-US-00015 TRAV30*01-CGTGTDSWGKLQF-TRAJ24*02 SEQ ID NO: 43 METLLKVL SGTLLWQLTW VRSQQPVQSP QAVILREGED AVINCSSSKA LYSVHWYRQK HGEAPVFLMI LLKGGEQKGH EKISASFNEK KQQSSLYLTA SQLSYSGTYF CGTGTDSWGK LQFGAGTQVV VTPD TRBV12-4*01-CASSPGLAGGEQFF-TRBD2*02-TRBJ2-1*01 SEQ ID NO: 44 MGSWTL CCVSLCILVA KHTDAGVIQS PRHEVTEMGQ EVTLRCKPIS GHDYLFWYRQ TMMRGLELLI YFNNNVPIDD SGMPEDRFSA KMPNASFSTL KIQPSEPRDS AVYFCASSPG LAGGEQFFGP GTRLTVL

Example 8: T-Cell Receptor T8 Recognizing HPV 16-E7 Epitope TLGIVCPI

(39) TABLE-US-00016 Immunogenic CDR3 peptide/ SEQ TCR Antigen purification TCR sequence ID NO: T8 E7 TLGIVCPI TRAV22*01-CA 25 VEPNSGNTPLVF- TRAJ29*01 IFNγ- TRBV7-2*02 or 26 CAPTURE 03-CASSLIISY NEQFF-TRBJ2- 1*01

(40) Full length variable chain sequences (CDR3 sequences are underlined):

(41) TABLE-US-00017 TRAV22*01-CAVEPNSGNTPLVF-TRAJ29*01 SEQ ID NO: 45 MKRILGAL LGLLSAQVCC VRGIQVEQSP PDLILQEGAN STLRCNFSDS VNNLQWFHQN PWGQLINLFY IPSGTKQNGR LSATTVATER YSLLYISSSQ TTDSGVYFCA VEPNSGNTPL VFGKGTRLSV IAN TRBV7-2*02 or 03-CASSLIISYNEQFF-TRBJ2-1*01 SEQ ID NO: 46 MGTRLL FWVAFCLLGA YHTGAGVSQS PSNKVTEKGK DVELRCDPIS GHTALYWYRQ RLGQGLEFLI YFQGNSAPDK SGLPSDRFSA ERTGESVSTL TIQRTQQEDS AVYLCASSLI ISYNEQFFGP GTRLTVL

Example 9: T-Cell Receptor T9 Recognizing HPV 16-E7 Epitope TLGIVCPI

(42) TABLE-US-00018 Immunogenic CDR3 peptide/ SEQ TCR Antigen purification TCR sequence ID NO: T9 E7 TLGIVCPI TRAV38-2/DV- 27 8*01-CAYRSAP YSGAGSYQLTF- TRAJ28*01 IFNγ- TRBV4-2*01-C 28 CAPTURE ASSQAPGLAGAE QYF-TRBD2*02- TRBJ2-7*01

(43) Full length variable chain sequences (CDR3 sequences are underlined):

(44) TABLE-US-00019 TRAV38-2/DV-8*01-CAYRSAPYSGAGSYQLTF-TRAJ28*01 SEQ ID NO: 47 MACPGFL WALVISTCLE FSMAQTVTQS QPEMSVQEAE TVTLSCTYDT SESDYYLFWY KQPPSRQMIL VIRQEAYKQQ NATENRFSVN FQKAAKSFSL KISDSQLGDA AMYFCAYRSA PYSGAGSYQL TFGKGTKLSV IPN TRBV4-2*01-CASSQAPGLAGAEQYF-TRBD2*02-TRBJ2-7*01 SEQ ID NO: 48 MGCRLL CCAVLCLLGA VPMETGVTQT PRHLVMGMTN KKSLKCEQHL GHNAMYWYKQ SAKKPLELMF VYNFKEQTEN NSVPSRFSPE CPNSSHLFLH LHTLQPEDSA LYLCASSQAP GLAGAEQYFG PGTRLTVT

Example 10: Interferon γ Release by TCR T2, T3 and T4 Transduced Cell Lines

(45) FIGS. 6 and 7 show that T-cell lines transduced with the TCR T3 and T4 (FIG. 6) and T2 (FIG. 7). Results are shown in the figures and figure legend.