Activation and Expansion of T Cells
The present disclosure relates to compositions, methods, and systems for selectively activating and/or expanding T cells for use in therapy. For example, the method may include contacting a population of T cells with an agent capable of binding an extracellular domain of a chimeric antigen receptor (CAR) that is expressed on the surface of the population of T cells.
1 . A method of selectively ex vivo activating a population of T cells for use in therapy, the method comprising:
contacting the population of T cells with an agent to activate the population of T cells that express a CAR to release IFN gamma, the agent comprising an extracellular domain of an antigen, the CAR comprising an extracellular domain, a transmembrane domain, an intracellular domain.
2 . The method of claim 1 , wherein the antigen comprises HER2, CD19, CD20, CD22, Kappa or light chain, CD30, CD33, CD123, CD38, ROR1, ErbB3/4, EGFR, EGFRvIII, EphA2, FAP, carcinoembryonic antigen, EGP2, EGP40, mesothelin, TAG72, PSMA, NKG2D ligands, B7-H6, IL-13 receptor α2, IL-11 receptor a, MUC1, MUC16, CA9, GD2, GD3, HMW-MAA, CD171, Lewis Y, G250/CAIX, HLA-AI MAGE A1, HLA-A2 NY-ESO-1, PSC1, folate receptor-α, CD44v7/8, 8H9, NCAM, VEGF receptors, 5T4, Fetal AchR, NKG2D ligands, CD44v6, TEM1, TEM8, or viral-associated antigens expressed by a tumor
3 . The method of claim 1 , further comprising:
prior to contacting the population of T cells with the agent,
(a) contacting the population of T cells with an anti-CD3 antibody that is immobilized on a solid surface for a time period;
(b) transferring a nucleic acid sequence encoding the CAR to the contacted population of T cells; and
(c) removing the solid surface from the population of T cells after the transferring.
4 . The method of claim 1 , further comprising:
collecting peripheral blood mononuclear cells (PBMCs) from a subject; and
selecting CD3 + cells from the PBMCs.
5 . The method of claim 1 , wherein the selecting the CD3 + cells from the PBMCs comprises:
mixing the PBMCs with a group of antibodies to allow the group of antibodies to bind target cells, the group of antibodies not including CD3 antibodies; and
removing the target cells from the PBMCs to obtain a solution containing the CD3 + cells.
6 . The method of claim 1 , wherein the group of antibodies comprises CD14, CD15, CD16, CD19, CD34, CD36, CD56, CD123, and CD235a.
7 . The method of claim 1 , wherein the antigen comprises the amino acid sequence of SEQ ID NO: 1.
8 . The method of claim 1 , wherein the agent is attached to a surface.
9 . The method of claim 8 , wherein the surface comprises a bead.
10 . The method of claim 9 , wherein the bead is capable of activating the population of T cells expressing the CAR to release IFN gamma.
11 . The method of claim 8 , wherein the surface comprises a magnetic particle.
12 . The method of claim 1 wherein the population of T cells is ex vivo expanded to a sufficient number for use in therapy.
13 . The method of claim 12 , wherein the population of T cells is expanded to at least 100-fold of the original T cell population.