LILRB1-based chimeric antigen receptor
Provided are chimeric antigen receptors having the hinge, transmembrane region, and/or intracellular domain of LILRB1, or functional fragments or variants thereof. Also provided herein are cells comprising the LILRB1 based receptors, and methods of making and using same.
1. A chimeric antigen receptor comprising a polypeptide comprising:
a) an antigen binding domain comprising an scFv comprising a CDR-L1, a CDR-L2, a CDR-L3, a CDR-H1, a CDR-H2, and a CDR-H3, wherein
i) the CDR-L1 comprises SEQ ID NO: 22, the CDR-L2 comprises SEQ ID NO: 23, the CDR-L3 comprises SEQ ID NO: 24, the CDR-H1 comprises SEQ ID NO: 25, the CDR-H2 comprises SEQ ID NO: 26, and the CDR-H3 comprises SEQ ID NO: 27; or
ii) the CDR-L1 comprises SEQ ID NO: 28, the CDR-L2 comprises SEQ ID NO: 29, the CDR-L3 comprises SEQ ID NO: 30, the CDR-H1 comprises SEQ ID NO: 31, the CDR-H2 comprises SEQ ID NO: 32, and the CDR-H3 comprises SEQ ID NO: 33;
b) an LILRB1 hinge domain comprising SEQ ID NO: 4, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 80, SEQ ID NO: 81, SEQ ID NO: 82, SEQ ID NO: 83, SEQ ID NO: 84, SEQ ID NO: 93, or a sequence having at least 95% identity thereto;
c) a transmembrane domain comprising SEQ ID NO: 5 or a sequence having at least 95% identity thereto; and
d) an LILRB1 intracellular domain comprising at least one immunoreceptor tyrosine-based inhibitory motif (ITIM) selected from the group consisting of NLYAAV (SEQ ID NO: 8), VTYAEV (SEQ ID NO: 9), VTYAQL (SEQ ID NO: 10), and SIYATL (SEQ ID NO: 11).
2. The receptor of claim 1 , wherein the intracellular domain comprises a sequence at least 95% identical to SEQ ID NO: 7.
3. The receptor of claim 1 , wherein the LILRB1 hinge and transmembrane comprise a sequence at least 95% identical to SEQ ID NO: 20.
4. The receptor of claim 1 , wherein the LILRB1 transmembrane and intracellular domain comprise a sequence at least 95% identical to SEQ ID NO: 21.
5. The receptor of claim 1 , wherein the LILRB1 hinge, transmembrane and intracellular domains comprise a sequence at least 95% identical to SEQ ID NO: 3.
6. The receptor of claim 1 , wherein the LILRB1 hinge, transmembrane and intracellular domains comprise a sequence at least 95% identical to SEQ ID NO: 2.
7. The receptor of claim 1 , wherein the LILRB1 hinge, transmembrane and intracellular domains comprise SEQ ID NO: 2.
8. The receptor of claim 1 , wherein the antigen binding domain specifically binds a HLA-A*02 allele of HLA-A.
9. The receptor of claim 1 , wherein the scFv comprises a sequence at least 95% identical to SEQ ID NO: 39 or SEQ ID NO: 125.
10. The receptor of claim 1 , wherein the receptor comprises an amino acid sequence at least 95% identical to SEQ ID NO: 122 or SEQ ID NO: 89.
11. The receptor of claim 1 , wherein the receptor comprises an amino acid sequence of SEQ ID NO: 122 or SEQ ID NO: 89.
12. A polynucleotide comprising a nucleic acid sequence encoding the polypeptide of claim 1 .
13. A vector comprising the polynucleotide of claim 12 .
14. An immune cell comprising the receptor of claim 1 .
15. The immune cell of claim 14 , wherein the immune cell is a T cell.
16. The immune cell of 15 , further comprising an activator receptor.