Methods and composition for producing and using immune cells and stem cells for cell-based therapies
Described herein are methods for selecting lymphocytes for adoptive cell therapy based on P-glycoprotein expression and compositions comprising same.
1. A method for isolating T cells and/or NK cells suitable for adoptive cell therapy, comprising:
(a) obtaining a sample comprising T cells and/or NK cells;
(b) optionally enriching the sample for T cells and/or NK cells; and
(c) isolating p-glycoprotein positive (Pgp+) T cells and/or NK from the sample, so as to obtain a fraction enriched in Pgp-positive T cells and/or NK cells by
(i) contacting the sample with at least one phototoxic compound that is a substrate of Pgp; and
(ii) exposing the sample to a light source sufficient to activate the at least one phototoxic compound so as to kill Pgp-negative cells,
thereby isolating T cells and/or NK cells suitable for adoptive cell transfer therapy.
2. The method of claim 1 , wherein the cells are contacted with the at least one phototoxic compound for a time and at a temperature sufficient to allow efflux of the phototoxic compound.
3. The method of claim 1 , wherein after contact with the at least one phototoxic compound the cells are removed from the photototoxic compound for a time and at a temperature sufficient to allow efflux of the phototoxic compound.
4. The method of claim 2 , wherein the temperature is 37° C.
5. The method of claim 1 , wherein the at least one phototoxic compound is any one or more of 2-(4,5-dibromo-6-amino-3-imino-3H-xanthen-9-yl)-benzoic acid methyl ester hydrochloride, 2-(4,5-dibromo-6-amino-3-imino-3H-xanthen-9-yl)-benzoic acid ethyl ester hydrochloride, 2-(4,5-dibromo-6-amino-3-imino-3H-xanthen-9-yl)-benzoic acid octyl ester hydrochloride, 2-(4,5-dibromo-6-amino-3-imino-3H-xanthen-9-yl)-benzoic acid n-butyl ester hydrochloride, 2-(6-ethyl amino-3-ethyl imino-3H-xanthen-9-yl)-benzoic acid n-butyl ester hydrochloride, or derivatives thereof or combinations thereof.
6. The method of claim 1 , wherein the light source is a visible light source.
7. The method of claim 1 , wherein the Pgp+ cells are further enriched using other methods for Pgp enrichment.
8. The method of claims 1 , wherein the fraction enriched in Pgp+ cells contains less than (a) 50% Pgp− cells; b) 40% Pgp− cells; c) 30% Pgp− cells; d) 20% Pgp− cells; d) 10% Pgp− cells; e) 5% Pgp− cells; or f) 1% Pgp− n cells.
9. The method of claim 1 , wherein the Pgp+ cells are further genetically modified so as to obtain genetically modified Pgp+ cells.
10. The method of claim 9 , wherein the genetically modified Pgp+ cells are selected from the group consisting of T cells and/or NK cells.
11. The method of claim 1 , wherein Pgp+ T cells are further genetically modified to express at least one chimeric antigen receptor, T cell receptor, synthetic immune receptor, chimeric T cell receptor, or other genetic element so as to obtain genetically modified Pgp-positive T cells.