METHODS FOR STIMULATING PROLIFERATION OR DIFFERENTIATION OF AN IMMUNE CELL WITH A MULTI-CHAIN CHIMERIC POLYPEPTIDE
The present disclosure relates to the field of biotechnology, and more specifically, to single-chain and multi-chain chimeric polypeptides having a linker domain positioned between two target-binding domains that are useful for a variety of applications including, without limitation, stimulating an immune cell, inducing or increasing proliferation of an immune cell, inducing differentiation of an immune cell, or treating a subject in need thereof (e.g., a subject having cancer or an aging-related disease or condition).
1 . A method of activating or inducing differentiation or expansion of a memory T cell, the method comprising contacting a memory T cell in a liquid culture medium comprising:
(1) 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; and
(iii) a first domain of a pair of affinity domains;
(b) a second chimeric polypeptide comprising:
(i) a second domain of the pair of affinity domains; 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
(2) an effective amount of an IgG1 antibody construct comprising at least one antigen-binding domain that binds specifically to the soluble tissue factor domain.
2 . The method of claim 1 , wherein the immune cell was previously obtained from a subject.
3 . The method of claim 1 , wherein the memory T cell is a peripheral blood memory T cell selected from the group consisting of: a Th17 cell, a Th22 cell, a Th9 cell, a Th2 cell, a Th1 cell, a Th3 cell, a λδ T cell, an αβ T cell, a tumor-infiltrating T cell, an effector T cell, a CD8 + T cell, and a CD4 + T cell.
4 . The method of claim 1 , wherein the memory T cell has previously been genetically modified to express a chimeric antigen receptor or a recombinant T cell receptor.
5 . The method of claim 1 , wherein the method further comprises, after the contacting step, introducing into the memory T cell a nucleic acid encoding a chimeric antigen receptor or a recombinant T cell receptor.
6 . The method of claim 1 , wherein the first target-binding domain and the soluble tissue factor domain directly abut each other in the first chimeric polypeptide.
7 . The method of claim 1 , wherein the first chimeric polypeptide further comprises a linker sequence between the first target-binding domain and the soluble tissue factor domain in the first chimeric polypeptide.
8 . The method of claim 1 , wherein the soluble tissue factor domain and the first domain of the pair of affinity domains directly abut each other in the first chimeric polypeptide.
9 . The method of claim 1 , wherein the first chimeric polypeptide further comprises a linker sequence between the soluble tissue factor domain and the first domain of the pair of affinity domains in the first chimeric polypeptide.
10 . The method of claim 1 , wherein the second domain of the pair of affinity domains and the second target-binding domain directly abut each other in the second chimeric polypeptide.
11 . The method of claim 1 , wherein the second chimeric polypeptide further comprises a linker sequence between the second domain of the pair of affinity domains and the second target-binding domain in the second chimeric polypeptide.
12 . The method of claim 1 , wherein the soluble tissue factor domain is a soluble human tissue factor domain.
13 . The method of claim 1 , wherein the soluble tissue factor domain does not stimulate blood coagulation.
14 . The method of claim 1 , wherein the multi-chain chimeric polypeptide does not stimulate blood coagulation.
15 . The method of claim 1 , wherein the contacting step is performed for a period of about 2 hours to about 20 days.
16 . The method of claim 1 , wherein the liquid culture medium comprises the multi-chain chimeric polypeptide and the IgG1 antibody construct at a molar ratio of about 0.5:1 to about 2:1.
17 . The method of claim 1 , wherein:
the soluble tissue factor domain comprises a sequence that is at least 80% identical to SEQ ID NO: 1;
the first domain of the pair of affinity domains comprises a sequence that is at least 80% identical to SEQ ID NO: 39;
the second domain of the pair of affinity domains comprises a sequence that is at least 80% identical to SEQ ID NO: 10; and
wherein:
(A) the first target-binding domain comprises a sequence that is at least 80% identical to SEQ ID NO: 135 and the second target-binding domain comprises a sequence that is at least 80% identical to SEQ ID NO: 124;
(B) the first target-binding domain comprises a sequence that is at least 80% identical to SEQ ID NO: 124 and the second target-binding domain comprises a sequence that is at least 80% identical to SEQ ID NO: 135;
(C) the first target-binding domain comprises a sequence that is at least 80% identical to SEQ ID NO: 124 and the second target-binding domain comprises a sequence that is at least 80% identical to SEQ ID NO: 60; or
(D) the first target-binding domain comprises a sequence that is at least 80% identical to SEQ ID NO: 60 and the second target-binding domain comprises a sequence that is at least 80% identical to SEQ ID NO: 60.
18 . The method of claim 17 , wherein:
the first target-binding domain comprises a sequence that is at least 80% identical to SEQ ID NO: 135; and
the second target-binding domain comprises a sequence that is at least 80% identical to SEQ ID NO: 124.
19 . The method of claim 18 , wherein:
the first target-binding domain comprises a sequence that is at least 90% identical to SEQ ID NO: 135;
the soluble tissue factor domain comprises a sequence that is at least 90% identical to SEQ ID NO: 1;
the first domain of the pair of affinity domains comprises a sequence that is at least 90% identical to SEQ ID NO: 39;
the second domain of the pair of affinity domains comprises a sequence that is at least 90% identical to SEQ ID NO: 10; and
the second target-binding domain comprises a sequence that is at least 90% identical to SEQ ID NO: 124.
20 . The method of claim 18 , wherein:
the first target-binding domain comprises a sequence that is at least 95% identical to SEQ ID NO: 135;
the soluble tissue factor domain comprises a sequence that is at least 95% identical to SEQ ID NO: 1;
the first domain of the pair of affinity domains comprises a sequence that is at least 95% identical to SEQ ID NO: 39;
the second domain of the pair of affinity domains comprises a sequence that is at least 95% identical to SEQ ID NO: 10; and
the second target-binding domain comprises a sequence that is at least 95% identical to SEQ ID NO: 124.
21 . The method of claim 18 , wherein:
the first target-binding domain comprises SEQ ID NO: 135;
the soluble tissue factor domain comprises SEQ ID NO: 1;
the first domain of the pair of affinity domains comprises SEQ ID NO: 39;
the second domain of the pair of affinity domains comprises SEQ ID NO: 10; and
the second target-binding domain comprises SEQ ID NO: 124.
22 . The method of claim 18 , wherein:
the first chimeric polypeptide comprises a sequence at least 80% identical to SEQ ID NO: 141; and
the second chimeric polypeptide comprises a sequence at least 80% identical to SEQ ID NO: 145.
23 . The method of claim 18 , wherein:
the first chimeric polypeptide comprises a sequence at least 90% identical to SEQ ID NO: 141; and
the second chimeric polypeptide comprises a sequence at least 90% identical to SEQ ID NO: 145.
24 . The method of claim 18 , wherein:
the first chimeric polypeptide comprises a sequence at least 95% identical to SEQ ID NO: 141; and
the second chimeric polypeptide comprises a sequence at least 95% identical to SEQ ID NO: 145.
25 . The method of claim 18 , wherein:
the first chimeric polypeptide comprises SEQ ID NO: 141; and
the second chimeric polypeptide comprises SEQ ID NO: 145.
26 . The method of claim 17 , wherein:
the first target-binding domain comprises a sequence that is at least 80% identical to SEQ ID NO: 124; and
the second target-binding domain comprises a sequence that is at least 80% identical to SEQ ID NO: 135.
27 . The method of claim 26 , wherein:
the first target-binding domain comprises a sequence that is at least 90% identical to SEQ ID NO: 124;
the soluble tissue factor domain comprises a sequence that is at least 90% identical to SEQ ID NO: 1;
the first domain of the pair of affinity domains comprises a sequence that is at least 90% identical to SEQ ID NO: 39;
the second domain of the pair of affinity domains comprises a sequence that is at least 90% identical to SEQ ID NO: 10; and
the second target-binding domain comprises a sequence that is at least 90% identical to SEQ ID NO: 135.
28 . The method of claim 26 , wherein:
the first target-binding domain comprises a sequence that is at least 95% identical to SEQ ID NO: 124;
the soluble tissue factor domain comprises a sequence that is at least 95% identical to SEQ ID NO: 1;
the first domain of the pair of affinity domains comprises a sequence that is at least 95% identical to SEQ ID NO: 39;
the second domain of the pair of affinity domains comprises a sequence that is at least 95% identical to SEQ ID NO: 10; and
the second target-binding domain comprises a sequence that is at least 95% identical to SEQ ID NO: 135.
29 . The method of claim 26 , wherein:
the first target-binding domain comprises SEQ ID NO: 124;
the soluble tissue factor domain comprises SEQ ID NO: 1;
the first domain of the pair of affinity domains comprises SEQ ID NO: 39;
the second domain of the pair of affinity domains comprises SEQ ID NO: 10; and
the second target-binding domain comprises SEQ ID NO: 135.
30 . The method of claim 26 , wherein:
the first chimeric polypeptide comprises a sequence that is at least 80% identical to SEQ ID NO: 126; and
the second chimeric polypeptide comprises a sequence that is at least 80% identical to SEQ ID NO: 137.
31 . The method of claim 26 , wherein:
the first chimeric polypeptide comprises a sequence that is at least 90% identical to SEQ ID NO: 126; and
the second chimeric polypeptide comprises a sequence that is at least 90% identical to SEQ ID NO: 137.
32 . The method of claim 26 , wherein:
the first chimeric polypeptide comprises a sequence that is at least 95% identical to SEQ ID NO: 126; and
the second chimeric polypeptide comprises a sequence that is at least 95% identical to SEQ ID NO: 137.
33 . The method of claim 26 , wherein:
the first chimeric polypeptide comprises SEQ ID NO: 126; and
the second chimeric polypeptide comprises SEQ ID NO: 137.
34 . The method of claim 17 , wherein:
the first target-binding domain comprises a sequence that is at least 80% identical to SEQ ID NO: 124; and
the second target-binding domain comprises a sequence that is at least 80% identical to SEQ ID NO: 60.
35 . The method of claim 34 , wherein:
the first target-binding domain comprises a sequence that is at least 90% identical to SEQ ID NO: 124;
the soluble tissue factor domain comprises a sequence that is at least 90% identical to SEQ ID NO: 1;
the first domain of the pair of affinity domains comprises a sequence that is at least 90% identical to SEQ ID NO: 39;
the second domain of the pair of affinity domains comprises a sequence that is at least 90% identical to SEQ ID NO: 10; and
the second target-binding domain comprises a sequence that is at least 90% identical to SEQ ID NO: 60.
36 . The method of claim 34 , wherein:
the first target-binding domain comprises a sequence that is at least 95% identical to SEQ ID NO: 124;
the soluble tissue factor domain comprises a sequence that is at least 95% identical to SEQ ID NO: 1;
the first domain of the pair of affinity domains comprises a sequence that is at least 95% identical to SEQ ID NO: 39;
the second domain of the pair of affinity domains comprises a sequence that is at least 95% identical to SEQ ID NO: 10; and
the second target-binding domain comprises a sequence that is at least 95% identical to SEQ ID NO: 60.
37 . The method of claim 34 , wherein:
the first target-binding domain comprises SEQ ID NO: 124;
the soluble tissue factor domain comprises SEQ ID NO: 1;
the first domain of the pair of affinity domains comprises SEQ ID NO: 39;
the second domain of the pair of affinity domains comprises SEQ ID NO: 10; and
the second target-binding domain comprises SEQ ID NO: 60.
38 . The method of claim 34 , wherein:
the first chimeric polypeptide comprises a sequence that is at least 80% identical to SEQ ID NO: 126; and
the second chimeric polypeptide comprises a sequence that is at least 80% identical to SEQ ID NO: 132.
39 . The method of claim 34 , wherein:
the first chimeric polypeptide comprises a sequence that is at least 90% identical to SEQ ID NO: 126; and
the second chimeric polypeptide comprises a sequence that is at least 90% identical to SEQ ID NO: 132.
40 . The method of claim 34 , wherein:
the first chimeric polypeptide comprises a sequence that is at least 95% identical to SEQ ID NO: 126; and
the second chimeric polypeptide comprises a sequence that is at least 95% identical to SEQ ID NO: 132.
41 . The method of claim 34 , wherein:
the first chimeric polypeptide comprises SEQ ID NO: 126; and
the second chimeric polypeptide comprises SEQ ID NO: 132.
42 . The method of claim 17 , wherein:
the first target-binding domain comprises a sequence that is at least 80% identical to SEQ ID NO: 60; and
the second target-binding domain comprises a sequence that is at least 80% identical to SEQ ID NO: 60.
43 . The method of claim 42 , wherein:
the first target-binding domain comprises a sequence that is at least 90% identical to SEQ ID NO: 60;
the soluble tissue factor domain comprises a sequence that is at least 90% identical to SEQ ID NO: 1;
the first domain of the pair of affinity domains comprises a sequence that is at least 90% identical to SEQ ID NO: 39;
the second domain of the pair of affinity domains comprises a sequence that is at least 90% identical to SEQ ID NO: 10; and
the second target-binding domain comprises a sequence that is at least 90% identical to SEQ ID NO: 60.
44 . The method of claim 42 , wherein:
the first target-binding domain comprises a sequence that is at least 95% identical to SEQ ID NO: 160;
the soluble tissue factor domain comprises a sequence that is at least 95% identical to SEQ ID NO: 1;
the first domain of the pair of affinity domains comprises a sequence that is at least 95% identical to SEQ ID NO: 39;
the second domain of the pair of affinity domains comprises a sequence that is at least 95% identical to SEQ ID NO: 10; and
the second target-binding domain comprises a sequence that is at least 95% identical to SEQ ID NO: 60.
45 . The method of claim 42 , wherein:
the first target-binding domain comprises SEQ ID NO: 60;
the soluble tissue factor domain comprises SEQ ID NO: 1;
the first domain of the pair of affinity domains comprises SEQ ID NO: 39;
the second domain of the pair of affinity domains comprises SEQ ID NO: 10; and
the second target-binding domain comprises SEQ ID NO: 60.
46 . The method of claim 42 , wherein:
the first chimeric polypeptide comprises a sequence that is at least 80% identical to SEQ ID NO: 161; and
the second chimeric polypeptide comprises a sequence that is at least 80% identical to SEQ ID NO: 130.
47 . The method of claim 42 , wherein:
the first chimeric polypeptide comprises a sequence that is at least 90% identical to SEQ ID NO: 161; and
the second chimeric polypeptide comprises a sequence that is at least 90% identical to SEQ ID NO: 130.
48 . The method of claim 42 , wherein:
the first chimeric polypeptide comprises a sequence that is at least 95% identical to SEQ ID NO: 161; and
the second chimeric polypeptide comprises a sequence that is at least 95% identical to SEQ ID NO: 130.
49 . The method of claim 42 , wherein:
the first chimeric polypeptide comprises SEQ ID NO: 161; and
the second chimeric polypeptide comprises SEQ ID NO: 130.