Ex vivo development, expansion and in vivo analysis of a novel lineage of dendritic cells

Abstract

Disclosed herein are new methods of producing a novel line of dendritic cells. The method comprises subjecting a sample of hematopoietic stem/precursor cells to a first feeder culture system that is supplemented with a first set of factors and a second feeder culture system supplemented with a second group of factors. The disclosure also pertains to new cell types that may be used as cancer immunotherapy.

What is claimed is: 
     
         1 . A method of producing a lineage of DC cells, said method comprising:
 culturing a population of hematopoietic stem/progenitor cells (HPCs) in a first feeder culture supplemented with kit ligand (KL); fms-like tyrosine kinase 3 ligand (FL); thrombopoietin (TPO); IL-3; IL-6 and/or basic fibroblast growth factor (bFGF) under conditions to produce a population of first expanded cells;   culturing said first expanded cells in a second feeder culture supplemented with KL, FL, TPO, GM-CSF, and/or IL-15 under conditions to produce DC progenitor cells (DCPs).   
     
     
         2 . The method of  claim 1 , wherein the first feeder culture comprises cells engineered to produce KL, FL, TPO, IL-3, IL-6 and/or bFGF. 
     
     
         3 . The method of  claim 2 , wherein said first feeder culture comprises cells engineered via a viral vector comprising an expression construct comprising a sequence encoding KL, FL, TPO, IL-3, IL-6 and/or bFGF. 
     
     
         4 . The method of  claim 1 , wherein the second feeder culture comprises cells engineered to produce KL, FL, TPO, GM-CSF, and/or IL-15. 
     
     
         5 . The method of  claim 1 , wherein said first expanded cells possess HPC phenotypic characteristics. 
     
     
         6 . The method of  claim 1 , further comprising culturing said DC progenitor cells in a third culture supplemented with GM-CSF and IL-15 under conditions to produce a population of cells having a phenotype similar to myeloid DCs. 
     
     
         7 . The method of  claim 6 , wherein the third culture is feeder free. 
     
     
         8 . A sample of DCPs produced by the method of  claim 1 . 
     
     
         9 . A sample of cells produced by the method of  claim 6 . 
     
     
         10 . A method of treating a patient in need, said method comprising administering a therapeutically effective amount of a sample of cells according to  claim 6 , wherein said patient in need is one who has cancer or infection related condition. 
     
     
         11 . The method of  claim 5 , wherein said patient in need is one who has multiple myeloma, acute myeloid leukemia, acute lymphoblastic leukemia, Hodgkin's lymphoma or glioblastoma. 
     
     
         12 . The method of  claim 5 , wherein said cells are autologous to or compatible with said patient in need. 
     
     
         13 . A method of treating a patient in need, said method comprising administering a therapeutically effective amount of a sample of cells according to  claim 9 , wherein said patient in need is one who has cancer or infection related condition. 
     
     
         14 . The method of  claim 8 , wherein said patient in need is one who has multiple myeloma, acute myeloid leukemia, acute lymphoblastic leukemia, Hodgkin's lymphoma orglioblastoma. 
     
     
         15 . The method of  claim 13 , wherein said HPCs are autologous to or compatible with said patient in need.