Determining WT-1 specific T cells and WT-1 specific T cell receptors (TCRs)
The invention is directed to methods for determining antigen-specific T cells generally and to T cell receptors which bind an epitope of the Wilms' tumor antigen-1 (WT1) protein specifically. The disclosure also provides polynucleotides encoding the TCRs, engineered cells exogenously expressing the TCRs, and methods of making and using the TCRs and/or cells expressing the TCRs.
1. A method for treating cancer in a subject in need thereof, the method comprising:
administering to the subject a therapeutically effective amount of a pharmaceutical composition comprising a recombinant T cell receptor (TCR) that binds Wilms' tumor antigen-1 (WT1), wherein the recombinant TCR comprises an alpha chain and a beta chain, wherein the alpha chain comprises a CDR3 sequence comprising an amino acid sequence having at least 90% sequence identity to an amino acid sequence set forth in SEQ ID NOs: 3, 11, 19, 27, 35, or 43, wherein the beta chain comprises a CDR3 sequence comprising an amino acid sequence having at least 90% sequence identity to an amino acid sequence set forth in SEQ ID NOs: 6, 14, 22, 30, 38, or 46, and wherein the cancer is treated by such administration.
2. The method according to claim 1 , wherein the beta chain comprises a CDR2 sequence comprising an amino acid sequence having at least 90% sequence identity to an amino acid sequence set forth in SEQ ID NOs: 5, 13, 21, 29, 37, or 45.
3. The method according to claim 2 , wherein the alpha chain comprises a CDR2 sequence comprising an amino acid sequence having at least 90% sequence identity to an amino acid sequence set forth in SEQ ID NOs: 2, 10, 18, 26, 34, or 42.
4. The method according to claim 1 , wherein the beta chain comprises a CDR1 sequence comprising an amino acid sequence having at least 90% sequence identity to an amino acid sequence set forth in SEQ ID NOs: 4, 12, 20, 28, 36, or 44.
5. The method according to claim 4 , wherein the alpha chain comprises a CDR1 sequence comprising an amino acid sequence having at least 90% sequence identity to an amino acid sequence set forth in SEQ ID NOs: 1, 9, 17, 25, 33, or 41.
6. The method according to claim 1 , wherein the TCR comprises:
(a) an alpha chain CDR3 comprising an amino acid sequence according to SEQ ID NO: 3 and a beta chain CDR3 comprising an amino acid sequence according to SEQ ID NO: 6; or
(b) an alpha chain CDR3 comprising an amino acid sequence according to SEQ ID NO: 11 and a beta chain CDR3 comprising an amino acid sequence according to SEQ ID NO: 14;
(c) an alpha chain CDR3 comprising an amino acid sequence according to SEQ ID NO: 19 or SEQ ID NO: 23, and a beta chain CDR3 comprising an amino acid sequence according to SEQ ID NO: 22;
(d) an alpha chain CDR3 comprising an amino acid sequence according to SEQ ID NO: 27 or SEQ ID NO: 31, and a beta chain CDR3 comprising an amino acid sequence according to SEQ ID NO: 30;
(e) an alpha chain CDR3 comprising an amino acid sequence according to SEQ ID NO: 35 and a beta chain CDR3 comprising an amino acid sequence according to SEQ ID NO: 38; or
(f) an alpha chain CDR3 comprising an amino acid sequence according to SEQ ID NO: 43 and a beta chain CDR3 comprising an amino acid sequence according to SEQ ID NO: 46.
7. A method for treating cancer in a subject in need thereof, comprising the steps:
a) isolating T cells from peripheral blood mononuclear cells (PBMC) obtained from the subject;
b) transducing or transfecting the T cells with a nucleic acid comprising a sequence selected from the group consisting of SEQ ID NOs: 55, 56, 63, 64, 71, 72, 79, 80, 87, 88, 95, and/or 96; and
c) administering the transduced or transfected T cells to the subject under conditions permissive for expression of the nucleic acid from the T cells in the subject.
8. The method according to claim 7 , wherein the T cells are further transduced or transfected with a nucleic acid encoding an exogenous immune activating molecule.
9. The method according to claim 8 , wherein the exogenous immune activating molecule is a cytokine or a ligand.
10. The method according to claim 9 , wherein the exogenous immune activating molecule is selected from the group consisting of: IL-12, TNFα, IFNγ, IL2, IL-7, IL-15, IL-18, CD40L, and IL-21.