METHODS OF GENERATING, STIMULATING, OR INCREASING PROLIFERATION OF DIFFERENT SUBSETS OF CD8+ T CELLS
Provided herein are multi-chain chimeric polypeptides and use thereof in generating, and/or stimulating, and/or increasing proliferation of different subsets of CD8 + T cells. Further, provided herein are pharmaceutical compositions comprising the multi-chain chimeric polypeptides. In addition, kits comprising the pharmaceutical compositions comprising the multi-chain chimeric polypeptides disclosed herein are also provided.
1 . A method of increasing proliferation of a CD8 + T cell selected from the group consisting of a progenitor exhausted CD8 + T (Tpex) cell, an effector memory CD8 + T (Tem) cell, a central memory CD8 + T (Tcm) cell, a memory stem CD8 + T (Tscm) cell, an antigen-experienced CD8 + T cell, and a terminally-exhausted CD8 + T (Tex) cell in vitro, the method comprising:
contacting the CD8 + T cell with an effective amount of a multi-chain chimeric polypeptide comprising:
(a) a first chimeric polypeptide comprising:
(i) a first target-binding domain;
(ii) a soluble tissue factor domain comprising a sequence that is at least 90% identical to SEQ ID NO:1; and
(iii) a first domain of a pair of affinity domains comprising a sequence that is at least 90% identical to SEQ ID NO:24;
(b) a second chimeric polypeptide comprising:
(i) a second domain of a pair of affinity domains comprising a sequence that is at least 90% identical to SEQ ID NO:22; and
(ii) a second target-binding domain,
wherein:
the first chimeric polypeptide and the second chimeric polypeptide associate through the binding of the first domain and the second domain of the pair of affinity domains; and
the first target-binding domain and the second target-binding domain each comprises a soluble TGF-ß receptor II (TGF-ßRII) having an amino acid sequence that is at least 90% identical to SEQ ID NO:5.
2 . The method of claim 1 , wherein the method does not comprise specific-antigen stimulation.
3 - 10 . (canceled)
11 . The method of claim 1 , wherein the first chimeric polypeptide and/or the second chimeric polypeptide further comprises a signal sequence at its N-terminal end.
12 . The method of claim 1 , wherein the first target-binding domain comprises a sequence of SEQ ID NO:5.
13 . (canceled)
14 . The method of claim 1 , wherein the first chimeric polypeptide comprises a sequence that is at least 90% identical to SEQ ID NO: 8.
15 . The method of claim 14 , wherein the first chimeric polypeptide comprises a sequence of SEQ ID NO: 8.
16 . The method of claim 15 , wherein the first chimeric polypeptide comprises a sequence of SEQ ID NO:10.
17 . The method of claim 1 , wherein the second target-binding domain comprises a sequence of SEQ ID NO:5.
18 . (canceled)
19 . The method of claim 1 , wherein the second chimeric polypeptide comprises a sequence that is at least 90% identical to SEQ ID NO:12.
20 . The method of claim 19 , wherein the second chimeric polypeptide comprises a sequence of SEQ ID NO: 12.
21 . The method of claim 20 , wherein the second chimeric polypeptide comprises a sequence of SEQ ID NO:14.
22 . A pharmaceutical composition comprising a therapeutically effective amount of (i) a multi-chain chimeric polypeptide, (ii) a T-cell or a CAR-T cell, and optionally (iii) a T-cell stimulating agent,
wherein the multi-chain chimeric polypeptide comprises:
(a) a first chimeric polypeptide comprising:
(i) a first target-binding domain;
(ii) a soluble tissue factor domain comprising a sequence that is at least 90% identical to SEQ ID NO:1; and
(iii) a first domain of a pair of affinity domains comprising a sequence that is at least 90% identical to SEQ ID NO:24;
(b) a second chimeric polypeptide comprising:
(i) a second domain of a pair of affinity domains comprising a sequence that is at least 90% identical to SEQ ID NO:22; and
(ii) a second target-binding domain,
wherein:
the first chimeric polypeptide and the second chimeric polypeptide associate through the binding of the first domain and the second domain of the pair of affinity domains; and
the first target-binding domain and the second target-binding domain each comprise a soluble TGF-ß receptor II (TGF-ßRII) having an amino acid sequence that is at least 90% identical to SEQ ID NO:5.
23 . The pharmaceutical composition of claim 22 , wherein the pharmaceutical composition comprises a T-cell.
24 . The pharmaceutical composition of claim 22 , wherein the pharmaceutical composition comprises a CAR-T cell.
25 . The pharmaceutical composition of claim 22 , wherein the pharmaceutical composition comprises a T-cell stimulating agent.
26 - 28 . (canceled)
29 . The pharmaceutical composition of claim 22 , wherein the first target-binding domain comprises a sequence of SEQ ID NO:5.
30 . (canceled)
31 . The pharmaceutical composition of claim 22 , wherein the first chimeric polypeptide comprises a sequence that is at least 90% identical to SEQ ID NO:8.
32 . The pharmaceutical composition of claim 31 , wherein the first chimeric polypeptide comprises a sequence of SEQ ID NO:8.
33 . The pharmaceutical composition of claim 32 , wherein the first chimeric polypeptide comprises a sequence of SEQ ID NO:10.
34 . The pharmaceutical composition of claim 22 , wherein the second target-binding domain comprises a sequence of SEQ ID NO:5.
35 . (canceled)
36 . The pharmaceutical composition of claim 22 , wherein the second chimeric polypeptide comprises a sequence that is at least 90% identical to SEQ ID NO:12.
37 . The pharmaceutical composition of claim 36 , wherein the second chimeric polypeptide comprises a sequence of SEQ ID NO:12.
38 . The pharmaceutical composition of claim 37 , wherein the second chimeric polypeptide comprises a sequence of SEQ ID NO:14.
39 . A kit comprising the pharmaceutical composition of claim 22 .