Granted Patent
B2
US 12,668,778 · App. 18/146,142 · Granted Jun 30, 2026
Method for expansion of double negative regulatory T cells
Inventors:
Li Zhang (Toronto, CA); Paulina Achita (Mississauga, CA); Jongbok Lee (Toronto, CA); Dalam Ly (Toronto, CA); Dzana Dervovic (Toronto, CA)
Assignee:
UNIVERSITY HEALTH NETWORK
C12N5/0637A61K40/11A61K40/22A61K40/418A61K40/42A61P35/00A61K40/50A61K2239/31A61K2239/38A61K2239/48C12N2501/51C12N2501/515C12N2501/999
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Abstract
There is provided herein a method for expanding human CD4 − CD8 − regulatory T cells (DN Tregs) from a population of cells comprising DN Tregs, comprising: culturing the population of cells with artificial antigen presenting cells (APCs), preferably the DN Tregs are αβ-TCR + CD56 − or alternatively γδ-TCR+.
Claims (36)
1 . A method of treating allograft rejection, graft vs. host disease (GVHD), or an autoimmune disease in a subject, comprising:
expanding human CD4 − CD8 − regulatory T cells (DN Tregs) from a population of cells comprising DN Tregs, by:
depleting the population of cells comprising DN Tregs of CD4 + and CD8 + cells;
activating the depleted population of cells with anti-CD3 antibodies; then
culturing the activated population of cells with artificial antigen presenting cells (APCs) that express at least one cell surface adhesion molecule for immunological synapse, the at least one cell surface adhesion molecule comprising at least one of CD54 and CD58;
wherein the DN Tregs are αβ-TCR + CD56 − , γδ-TCR + , or both; and
administering to the subject a therapeutically effective amount of the expanded DN Tregs.
2 . The method of claim 1 , for treating allograft rejection.
3 . The method of claim 1 , for treating GVHD.
4 . The method of claim 1 , for treating an autoimmune disease.
5 . The method of claim 1 , wherein the artificial APCs display anti-CD3 antibodies.
6 . The method of claim 5 , wherein the artificial APCs further express a co-stimulatory molecule.
7 . The method of claim 6 , wherein the co-stimulatory molecule is at least one of CD80, CD86, CD83, 4-1BBL, OX40L, ICOSL, CD40L, and CD28 antibody.
8 . The method of claim 5 , wherein the artificial APCs do not express inhibitory molecules.
9 . The method of claim 5 , wherein the artificial APCs further express at least one of M-CSF, IL-6, IL-8, TGF-β, MIP-1a, IL-7, IL-15 and IL-2.
10 . The method of claim 1 , wherein the artificial APCs are K-562 cells.
11 . The method of claim 1 , wherein the culturing is additionally in the presence of at least one of IL-2, IL-7 and IL15.
12 . The method of claim 1 , wherein the activating is with anti-CD3 antibodies cross-linked, or otherwise attached, to a surface.
13 . The method of claim 1 , wherein the activating is with soluble anti-CD3 antibodies.
14 . The method of claim 1 , wherein the activating is sequentially with anti-CD3 antibodies cross-linked, or otherwise attached, to a surface and soluble anti-CD3 antibodies.
15 . The method of claim 1 , wherein the population comprises peripheral blood mononuclear cells (PBMCs) or cord blood mononuclear cells (CMBCs).
16 . The method of claim 1 , further comprising incubating the expanded DN Tregs with at least one inhibitor of the PI3K/AKT/mTOR pathway prior to administration.
17 . The method of claim 16 , wherein the at least one inhibitor is a mTOR inhibitor, dual PI3K/mTOR inhibitor, AKT inhibitor, or Pan-class I and isoform-specific PI3K inhibitors.
18 . The method of claim 16 wherein the inhibitor is a Rapalog.
19 . The method of claim 16 , wherein the inhibitor is rapamycin, deforolimus, emsirolimus, everolimus, ridaforolimus, temsirolimus or a mTORC1/mTORC2 dual inhibitor.
20 . The method of claim 19 , wherein the inhibitor is rapamycin.
21 . The method of claim 16 , wherein the inhibitor is Wortmannin, LY294002, PKI-179, or Akt inhibitor IV.
22 . The method of claim 8 , wherein the inhibitory molecule is PDL1, PDL2, B7H3, or B7H4.
23 . The method of claim 1 , wherein the artificial APC is an artificial APC:
i. expressing 4-1BBL and membrane-bound IL-15;
ii. expressing CD64, CD86, 4-1BBL, truncated CD19, and membrane-bound IL-21;
iii. expressing CD64 and CD86;
iv. expressing HLA class I, CD80, and CD83;
v. expressing CD64 and 4-1BBL;
vi. expressing CD32, CD80, CD83, CD86, and 4-1BBL; or
vii. expressing CD64, CD86, 4-1BBL, truncated CD19, and membrane-bound IL-15.
Assignments (1)
ASSIGNMENT OF ASSIGNOR'S INTEREST
Recorded Dec 23, 2022
From: ZHANG, LI; ACHITA, PAULINA; LEE, JONG BOK; LY, DALAM; DERVOVIC, DZANA
To: UNIVERSITY HEALTH NETWORK
Reel/Frame 062196/0802 →
Continuity (4)
Division
15573379
Provisional Application
62237050
· Oct 5, 2015
Provisional Application
62159561
· May 11, 2015
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