Engineered immune-mobilizing T-cell receptors with enhanced affinity for HIV-1 Gag
The present invention relates to T cell receptors (TCRs) which bind the HLA-A*02 restricted peptide SLYNTVATL (SEQ ID NO: 1) derived from the HIV Gag gene product, p17. Said TCRs comprise non-natural mutations within the alpha and/or beta variable domains relative to a native HIV TCR. The TCRs of the invention possess unexpectedly high affinity, specificity and sensitivity for a complex of SEQ ID NO: 1 and HLA-A*02, and drive a particularly potent T cell response. Such TCRs are particularly useful in the development of soluble immunotherapeutic reagents for the treatment of HIV infected individuals.
1. A T-cell receptor (TCR) having the property of binding to SLYNTVATL (SEQ ID No: 1) in complex with HLA-A*02, comprising:
a TCR alpha chain variable domain and
a TCR beta chain variable domain, wherein
the alpha chain variable domain comprises the amino acid sequence set forth in SEQ ID NO: 6 or 7, and
the beta chain variable domain comprises the amino acid sequence set forth in SEQ ID NO: 8.
2. The TCR of claim 1 , wherein the TCR is an alpha-beta heterodimer having an alpha chain TRAC constant domain sequence and a beta chain TRBC1 or TRBC2 constant domain sequence.
3. The TCR of claim 2 , wherein the alpha and beta chain constant domain sequences are modified by truncation or substitution to delete the native disulphide bond between Cys4 of exon 2 of TRAC and Cys2 of exon 2 of TRBC1 or TRBC2.
4. The TCR of claim 2 , wherein the alpha and beta chain constant domain sequence(s) are modified by substitution of cysteine residues for Thr 48 of TRAC and Ser 57 of TRBC1 or TRBC2, the said cysteines forming a disulphide bond between the alpha and beta constant domains of the TCR.
5. The TCR of claim 1 , wherein the TCR has a single chain format of the type Va-L-Vb, Vb-L-Va, Va-Ca-L-Vb, Va-L-Vb-Cb, Va-Ca-L-Vb-Cb or Vb-Cb-L-Va-Ca wherein Va and Vb are TCR alpha and beta variable regions respectively, Ca and Cb are TCR alpha and beta constant regions respectively, and L is a linker sequence.
6. The TCR of claim 1 , wherein the TCR is associated with a detectable label, a therapeutic agent or a PK modifying moiety.
7. The TCR of claim 1 , wherein the TCR is associated with an anti-CD3 antibody covalently linked to the C- or N-terminus of the alpha or beta chain of the TCR.
8. The TCR of claim 7 , wherein the anti-CD3 antibody is covalently linked to the C- or N-terminus of the beta chain of the TCR via a linker sequence.
9. The TCR of claim 8 , wherein the linker sequence is selected from the group consisting of: GGGGS (SEQ ID NO: 12), GGGSG (SEQ ID NO: 13), GGSGG (SEQ ID NO: 14), GSGGG (SEQ ID NO: 15), GSGGGP (SEQ ID NO: 16), GGEPS (SEQ ID NO: 17), GGEGGGP (SEQ ID NO: 18), and GGEGGGSEGGGS (SEQ ID NO: 19).
10. A pharmaceutical composition, comprising: a TCR of claim 1 ; and one or more pharmaceutically acceptable carriers or excipients.