Photodynamic process and product obtained therefrom
The present invention relates to a method for ex vivo photodynamic treatment of cells. A cell preparation is incubated in a first medium having 5 μM or less of a photosensitive compound. A population of cells in the cell preparation retains the photosensitive compound, for example, a salt of 2-(4,5-dibromo-6-amino-3-imino-3H-xanthen-9-yl)-benzoic acid methyl ester. The first medium is then replaced with an extrusion medium to remove the photosensitive compound. The cell preparation is then illuminated in the extrusion medium to selectively kill the population of cells retaining the photosensitive compound, thereby creating an illuminated cell preparation.
1. A method for ex vivo photodynamic treatment of cells, comprising the steps of:
incubating a cell preparation in a first medium comprising 5 μM of a photosensitive compound, wherein the cell preparation in the first medium has a first cell concentration, wherein a population of cells retains the photosensitive compound, wherein the photosensitive compound is a salt of 2-(4, 5-dibromo-6-amino-3-imino-3H-xanthen-9-yl)-benzoic acid methyl ester;
replacing the first medium with an extrusion medium to remove the photosensitive compound, wherein the cell preparation in the extrusion medium has a second cell concentration, and wherein the ratio of the first cell concentration to the second cell concentration is 1:3; and
illuminating the cell preparation in the extrusion medium to selectively kill the population of cells retaining the photosensitive compound, thereby creating an illuminated cell preparation.
2. The method of claim 1 , wherein the salt of 2-(4, 5-dibromo-6-amino-3-imino-3H-xanthen-9-yl)-benzoic acid methyl ester is a hydrobromide or hydrochloride salt.
3. The method of claim 1 , wherein the cell preparation in the medium comprising 5 μM of a photosensitive compound was first subjected to a mixed lymphocyte reaction.
4. The method of claim 1 , wherein the cell preparation in the medium comprising 5 μM of a photosensitive compound comprises T-lymphocytes from a first person and non-proliferative T-lymphocytes from a second person.
5. The method of claim 4 , wherein the first person is a donor and the second person is a recipient.
6. The method of claim 1 , wherein the population of cells that retains the photosensitive compound are recipient-activated donor cells.
7. The method of claim 6 , wherein the recipient-activated donor cells are alloreactive T-lymphocytes.
8. The method of claim 5 , further comprising the step of infusing the illuminated cell preparation into the second person.
9. The method of claim 5 , wherein the second person has cancer.
10. The method of claim 5 , wherein the second person is a transplant patient.
11. The method of claim 1 , the first medium is replaced with the extrusion medium using a separation method.
12. The method of claim 11 , wherein the separation method is selected from the group consisting of centrifugation, filtration, and dialysis.
13. The method of claim 1 , wherein during the illuminating step, the cells are presented as a thin layer of cells, wherein the thin layer is between 1.0 mm and 20 mm thick.
14. The method of claim 1 , wherein at least 80% of the population of cells retaining the photosensitive compound is killed during the illuminating step.