Devices and methods for selecting apoptosis-signaling resistant cells, and uses thereof

Abstract

The present invention discloses a device and a kit adapted for selection of cells that are resistant to receptor-mediated apoptosis and a method for using the device and kit. The device enables negative selection of mature immune cells which induce graft versus host disease (GvHD) out of a heterogeneous cell population which is introduced into the device. The device enables an efficient cell selection in simplified and cheaper setting by an off the shelf product—a solution that currently do not exist. The present invention further discloses uses for the device.

1 - 38 . (canceled) 
     
     
         39 . A device, comprising: a container comprised of a biocompatible material and a biologically active apoptosis-inducing ligand immobilized to a surface, wherein said container comprises two interlocking chambers separated by a filter apparatus adapted to separate whole cells from cell debris and proteins, and wherein said device is adapted for cell selection by apoptosis of apoptosis-sensitive cells and said apoptosis-inducing ligand is selected from the group consisting of tumor necrosis factor α (TNF-α), Fas ligand (FasL), Trail, Tweak, and any combination thereof. 
     
     
         40 . The device of  claim 39 , wherein said surface is the inner surface of said container. 
     
     
         41 . The device of  claim 39 , wherein said biocompatible material is selected from the group comprising: polypropylene, polystyrene, silicone, polyvinyl chloride or a combination thereof. 
     
     
         42 . The device of  claim 39 , wherein said immobilized apoptosis-inducing ligand is Fas ligand (FasL). 
     
     
         43 . A cell selection kit, comprising: (a) a device of  claim 39 ; and (b) a solution for maintaining the integrity and activity of said apoptosis-inducing ligand within said device. 
     
     
         44 . The kit of  claim 43 , further comprising a second apoptosis-inducing ligand. 
     
     
         45 . A method for selecting an apoptosis-signaling resistant cell from a cell population, said cell population comprises an apoptosis-signaling resistant cell and an apoptosis-signaling sensitive cell, said method comprising the steps of:
 (a) introducing a sample comprising a cell population into the device of  claim 39 ; and   (b) incubating said cells within said device;   thereby selecting for apoptosis-signaling resistant cells through enrichment of such apoptosis-signaling resistant cells in the cell population by inducing cell death in apoptosis-signaling sensitive cells to eliminate such apoptosis-signaling sensitive cells from the cell population.   
     
     
         46 . The method of  claim 45 , wherein said apoptosis-signaling resistant cell comprises: a stem cell, an immune cell insensitive to activation-induced cell death (AICD), a progenitor cell, or any combination thereof. 
     
     
         47 . The method of  claim 46 , wherein said stem cell is a bone marrow stem cell. 
     
     
         48 . The method of  claim 46 , wherein said immune cell insensitive to activation-induced cell death is a T cell. 
     
     
         49 . The method of  claim 45 , wherein said cell population is derived from: bone marrow, a progenitor cell, mobilized peripheral blood, or umbilical cord blood (UCB). 
     
     
         50 . The method of  claim 45 , wherein said cell-population is introduced into one chamber, incubated within the chamber and after incubation the filter is used to further isolate the selected cells from cell debris and proteins in the solution. 
     
     
         51 . A method for improving the clinical outcome of hematopoietic stem and progenitor cells (HSPC) transplantation, comprising the steps of:
 (a) providing a sample comprising a cell population, said cell population comprises stem and progenitor cell;   (b) contacting said cell population with a biologically active apoptosis-inducing ligand, Fas ligand (FasL) for a period of 1 to 24 hours or tumor necrosis factor α (TNF-α) for a period of 24 to 48 hours, in the device of  claim 39 ;   (c) retrieving the cells of step (b); and   (d) transplanting the cells of step (c);   thereby improving the clinical outcome of hematopoietic stem and progenitor cells (HSPC) transplantation.   
     
     
         52 . A method for eliminating a malignant cell in a composition comprising a progenitor-cell transplant, comprising the steps of:
 (a) providing a composition comprising a progenitor-cell transplant; and   (b) contacting said composition with an apoptosis-inducing ligand for a period of about 24 hours in the device of  claim 39 ;   thereby, eliminating a malignant cell in a composition comprising a progenitor-cell transplant.   
     
     
         53 . A method for preventing graft vs. host disease (GvHD) while retaining graft vs. tumor (GvT) activity, comprising the steps of:
 (a) providing a sample comprising a cell population, said cell population comprises HSPC and immune cells;   (b) contacting said cell population with an apoptosis-inducing ligand for 2-16 hours in the device of  claim 39 ;   (c) retrieving the cells of step (b); and   (d) transplanting the cells of step (c),   thereby preventing graft vs. host disease (GvHD) while retaining graft vs. tumor (GvT) activity.   
     
     
         54 . A device, comprising: a container comprised of a biocompatible material and a biologically active apoptosis-inducing ligand immobilized to a surface, wherein said container is in the form of a column, and wherein said device is adapted for cell selection by apoptosis of apoptosis-sensitive cells and said apoptosis-inducing ligand is selected from the group consisting of tumor necrosis factor α (TNF-α), Fas ligand (FasL), Trail, Tweak, and any combination thereof. 
     
     
         55 . The device of  claim 54 , wherein said biocompatible material is selected from the group comprising: polypropylene, polystyrene, silicone, polyvinyl chloride or a combination thereof. 
     
     
         56 . A cell selection kit, comprising: (a) a device of  claim 54 ; and (b) a solution for maintaining the integrity and activity of said apoptosis-inducing ligand within said device. 
     
     
         57 . A device, comprising: a container comprised of a biocompatible material and a biologically active apoptosis-inducing ligand immobilized to a surface, wherein said container comprises beads, and wherein said surface is the surface of beads present within said container, and wherein said device is adapted for cell selection by apoptosis of apoptosis-sensitive cells and said apoptosis-inducing ligand is selected from the group consisting of tumor necrosis factor α (TNF-α), Fas ligand (FasL), Trail, Tweak, and any combination thereof. 
     
     
         58 . A cell selection kit, comprising: (a) a device of  claim 57 ; and (b) a solution for maintaining the integrity and activity of said apoptosis-inducing ligand within said device.