H3.3 CTL peptides and uses thereof
Peptides that generate an immune response to glioma-related H3.3 proteins and methods of their use are provided.
1. A modified T-cell comprising a heterologous T-cell receptor (TCR), or fragment thereof, which binds to a peptide/major histocompatibility complex (MHC) complex, wherein the TCR or the fragment thereof binds to a histone H3 variant H3.3 peptide, wherein the TCR or fragment thereof comprises:
a TCR alpha chain or fragment thereof comprising complementarity determining regions (CDRs) 1, 2, and 3 comprising SEQ ID NOs: 12, 13, and 14, respectively; and
a TCR beta chain or fragment thereof comprising CDRs 1, 2, and 3 comprising SEQ ID NOs: 15, 16, and 17, respectively.
2. The T-cell of claim 1 , wherein the TCR alpha chain or fragment thereof is a TCR alpha chain and the TCR beta chain or fragment thereof is a TCR beta chain.
3. The T-cell of claim 2 , wherein the TCR alpha chain comprises the amino acid sequence set forth in SEQ ID NO: 8, and the TCR beta chain comprises the amino acid sequence set forth in SEQ ID NO: 10.
4. The T-cell of claim 3 , wherein the heterologous TCR is expressed from a nucleic acid comprising a polynucleotide sequence encoding a self-cleaving peptide that links polynucleotide sequences encoding the TCR alpha and beta chains.
5. The T-cell of claim 4 , wherein the self-cleaving peptide is a porcine teschovirus-1 2A (P2A) peptide.
6. The T-cell of claim 1 , wherein the heterologous TCR is expressed from a retroviral vector.
7. The T-cell of claim 6 , wherein the retroviral vector is a lentiviral vector.
8. The T-cell of claim 7 , wherein the T-cell exhibits downregulated expression of an endogenous T-cell receptor comprising an endogenous TCR alpha chain and an endogenous TCR beta chain.
9. The T-cell of claim 8 , wherein
i) the T-cell exhibits downregulated expression of an endogenous TCR alpha chain;
ii) the T-cell exhibits downregulated expression of an endogenous TCR beta chain; or
iii) the T-cell exhibits downregulated expression of an endogenous TCR alpha chain and downregulated expression of an endogenous TCR beta chain.
10. The T-cell of claim 9 , wherein
i) the T-cell exhibits downregulated expression of an endogenous TCR alpha chain due to expression of an siRNA complementary to the endogenous TCR alpha chain;
ii) the T-cell exhibits downregulated expression of an endogenous TCR beta chain due to expression of an siRNA complementary to the endogenous TCR beta chain; or
iii) the T-cell exhibits downregulated expression of an endogenous TCR alpha chain due to expression of an siRNA complementary to the endogenous TCR alpha chain and downregulated expression of an endogenous TCR beta chain due to expression of an siRNA complementary to the endogenous TCR beta chain.
11. The T-cell of claim 10 , wherein
i) the lentiviral vector further expresses the siRNA capable of downregulating expression of an endogenous TCR alpha chain;
ii) the lentiviral vector further expresses the siRNA capable of downregulating expression of an endogenous TCR beta chain; or
iii) the lentiviral vector further expresses the siRNA capable of downregulating expression of an endogenous TCR alpha chain and the siRNA capable of downregulating expression of an endogenous TCR beta chain.
12. The T-cell of claim 1 , wherein expression of the heterologous TCR is under the control of a heterologous promoter.
13. The T-cell of claim 12 , wherein the heterologous promoter is a constitutive promoter.
14. The T-cell of claim 11 , wherein expression of the siRNA capable of downregulating expression of an endogenous TCR alpha chain and expression of the siRNA capable of downregulating expression of an endogenous TCR beta chain are under the control of a heterologous promoter.
15. The T-cell of claim 1 , wherein the peptide is in a complex with a MHC.
16. The T-cell of claim 1 , wherein the peptide in the peptide/WIC complex comprises the amino acid sequence (R/A)MSAP(S/A)TGGV (SEQ ID NO: 1).
17. The T-cell of claim 16 , wherein the peptide in the peptide/MHC complex consists of 10-12 amino acids.
18. The T-cell of claim 17 , wherein the amino acid sequence is RMSAPSTGGV (SEQ ID NO: 2).