Granted Patent B2

US 12,516,290 · App. 17/777,746 · Granted Jan 6, 2026

Inventors: Myoung Ho Jang (Seoul, KR); Chun-Pyo Hong (Gyeonggi-do, KR); Young Joo Choi (Gyeonggi-do, KR); June Sub Lee (Gyeonggi-do, KR)

Assignee: GI CELL, INC.

C12N5/0636C12N2501/2302C12N2501/51C12N2506/115

Abstract

Claims (44)

1 . A method for culturing a T cell comprising:

culturing CD8+ T cells in a medium comprising a fusion protein dimer comprising an IL-2 variant and a CD80 protein or a fragment thereof,

wherein the fusion protein comprises the following structural formula (I) or (II):

N′-X-[linker (1)] n -Fc domain-[linker (2)] m -Y-C′  formula (I)

N′-Y-[linker (1)] n -Fc domain-[linker (2)] m -X-C′  formula (II)

wherein

N′ is the N-terminus of the fusion protein,

C′ is the C-terminus of the fusion protein,

X is a CD80 protein or a fragment thereof,

Y is the IL-2 variant,

the linkers (1) and (2) are peptide linkers, and

n and m are each independently 0 or 1,

wherein the IL-2 variant comprises any one selected from the following substitution combinations (a) to (d) in the amino acid sequence of SEQ ID NO: 10;

(a) R38A/F42A

(b) R38A/F42A/Y45A

(c) R38A/F42A/E61R

(d) R38A/F42A/L72G, and

wherein the CD80 protein fragment comprises an extracellular domain of CD80.

2 . The method according to claim 1 , wherein the CD8+ T cell is obtained from peripheral blood mononuclear cells (PBMCs).

3 . The method according to claim 1 , wherein the culturing is performed for 7 to 21 days.

4 . A method for in vitro activation and proliferation of CD8+ T cells comprising:

simultaneously culturing a cell and a cancer antigen in a medium comprising a fusion protein dimer comprising an IL-2 variant and a CD80 protein or a fragment thereof,

wherein the cell is either a peripheral blood mononuclear cell (PBMC) or a T cell,

wherein the fusion protein comprises the following structural formula (I) or (II):

N′-X-[linker (1)] n -Fc domain-[linker (2)] m -Y-C′  formula (I)

N′-Y-[linker (1)] n -Fc domain-[linker (2)] m -X-C′  formula (II)

wherein

N′ is the N-terminus of the fusion protein,

C′ is the C-terminus of the fusion protein,

X is a CD80 protein or a fragment thereof,

Y is the IL-2 variant,

the linkers (1) and (2) are peptide linkers, and

n and m are each independently 0 or 1,

wherein the IL-2 variant comprises any one selected from the following substitution combinations (a) to (d) in the amino acid sequence of SEQ ID NO: 10:

(a) R38A/F42A

(b) R38A/F42A/Y45A

(c) R38A/F42A/E61R

(d) R38A/F42A/L72G,

wherein the CD80 protein fragment comprises an extracellular domain of CD80.

5 . The method according to claim 4 , wherein the T cells are CD8+ T cells.

6 . The method according to claim 1 , wherein the IL-2 variant comprises the amino acid sequence of SEQ ID NO: 6, 22, 23, or 24.

7 . The method according to claim 1 , wherein the CD80 protein has the amino acid sequence of SEQ ID NO: 11.

8 . The method according to claim 1 , wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 9, 26, 28, or 30.

9 . The method according to claim 1 , wherein the T cells comprise memory T cells.

Assignments (1)

Priority Claims (1)

Continuity (1)

References Cited (69)