Artificial antigen presenting cells comprising protein L for expanding immune cells for immunotherapy
View Patent ↗Disclosed herein are methods of expanding immune cells for immunotherapy and/or increasing the purity of a population of CAR T cells using artificial antigen presenting cells (aAPCs) having on their surface Protein L. The disclosed aAPCs can also secrete antibodies that bind molecules of the T cell inhibitory pathway. For example, anti-CD3 scFv on the surface of the aAPCs can bind and activate T cells, while anti-CD28 scFv and 4-1BBL on the surface of the aAPCs can provide dual co-stimulation for the T cells resulting in decreased levels of the markers CD25, TIM3, LAG3, and PD1. For example, blocking PD1/PDL1 ligation can limit suppression that is mediated by the tumor microenvironment. This is a less costly and more efficient alternative to peripheral blood mononuclear cells (PBMCs) and cytokine treatments that result in better quality T cell for adoptive transfer back into patients.
1 . A method for expanding a γδ T cell from a subject for use in immune therapy, comprising:
(a) isolating a γδ T cell from peripheral blood of the subject,
b) providing an artificial antigen presenting cell (aAPC) comprising a cell membrane, wherein the aAPC expresses Protein L and comprises Protein L on its membrane; wherein the aAPC comprises one or more single chain variable fragment (scFv) antibodies or ligands that bind CD28; and
(c) contacting the peripheral blood-isolated γδ T cell with an effective amount of the aAPC to expand the γδ T cell in an amount effective for immunotherapy.
2 . The method of claim 1 , wherein the aAPC further expresses one or more scFv antibodies that bind a T cell inhibitory molecule, or a combination thereof.
3 . The method of claim 2 , wherein the T cell inhibitory molecule comprises PD1, PDL1, or a combination thereof.
4 . The method of claim 2 , wherein the T cell inhibitory molecule comprises CTLA4, LAG3, TIM3, BTLA, CD160, 2B4, A2aR, KIR, or any combination thereof.
5 . The method of claim 1 , wherein the cell membrane further comprises on its membrane one or more scFv or ligands that bind a second co-stimulatory molecule on T-cells, one or more scFv antibodies or ligands that selectively bind CD3, or a combination thereof.
6 . The method of claim 5 , wherein the co-stimulatory molecule comprises 4-1BB.
7 . A γδ T cell produced by the method of claim 1 .
8 . A method for increasing the purity of γδ T cells, comprising
(a) isolating a γδ T cell from peripheral blood of a subject:
(b) providing an artificial antigen presenting cell (aAPC) comprising a cell membrane, wherein the aAPC expresses Protein L and comprises Protein L on its membrane; wherein the aAPC comprises one or more scFv antibodies or ligands that bind CD28; and
(c) incubating the γδ T cell with the aAPC for at least 8 hours; wherein the aAPC induces the γδ T cellto proliferate.
9 . The method of claim 8 , wherein the γδ T cell is incubated with the aAPC for at least 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 26, 28, 30, 32, 34, 36, or 48 hours.
10 . The method of claim 8 , wherein the aAPC further expresses one or more scFv antibodies that bind a T cell inhibitory molecule, or a combination thereof.
11 . The method of claim 10 , wherein the T cell inhibitory molecule comprises PD1, PDL1, or a combination thereof.
12 . The method of claim 10 , wherein the T cell inhibitory molecule comprises CTLA4, LAG3, TIM3, BTLA, CD160, 2B4, A2aR, KIR, or any combination thereof.
13 . The method of claim 8 , wherein the cell membrane contains on its membrane one or more scFv antibodies or ligands that bind a co-stimulatory molecule on T-cells, one or more scFv antibodies or ligands that selectively bind CD3, or a combination thereof.
14 . The method of claim 13 , wherein the co-stimulatory molecule comprises 4-1BB.