Compositions and methods for producing dendritic cells

Abstract

Described herein are compositions and methods for treating a disease, particularly a cancer, with primed dendritic cells recognizing a tumor antigen. The methods may comprise storing, shipping and/or culturing dendritic cells, where the dendritic cells are stored on a hard surface. Lysis protocols are described where the lysis does not result in complete lysis of cells in order to provide cell surface molecules maintained in a cell surface-embedded state. Non-lethal Dengue virus strains are also provided for therapeutic purposes.

What is claimed is: 
     
         1 . A method for producing primed dendritic cells, comprising:
 a) culturing dendritic cells on a hard surface;   b) lysing at least one cell with a hypochlorite solution to produce a lysate;   c) contacting the dendritic cells with the lysate to produce primed dendritic cells; and   d) maturing the primed dendritic cells, wherein maturing comprises contacting the primed dendritic cells with a toll-like receptor 7 agonist or a toll-like receptor 8 agonist.   
     
     
         2 . The method of  claim 1 , wherein the primed dendritic cells produce at least 6 ng/mL IL-12p70. 
     
     
         3 . The method of  claim 1 , wherein the primed dendritic cells produce at least about 6.5 ng/mL IL-12p70. 
     
     
         4 . The method of  claim 1 , wherein the primed dendritic cells produce about 19 ng/mL IL-12p70. 
     
     
         5 . The method of  claim 1 , wherein the hard surface is a plastic surface. 
     
     
         6 . The method of  claim 1 , wherein the hard surface comprises polystyrene. 
     
     
         7 . The method of  claim 1 , wherein the hard surface is substantially free of a component that reduces a Type 1 response produced by the primed dendritic cells. 
     
     
         8 . The method of  claim 7 , wherein the component is selected from a fluorinated polyethylene, a fluorinated polypropylene, and a phthalate. 
     
     
         9 . The method of  claim 1 , wherein the lysate comprises a plurality of intact cells. 
     
     
         10 . The method of  claim 1 , wherein the toll-like receptor 7 agonist or toll-like receptor 8 agonist is an imidazoquinoline compound. 
     
     
         11 . The method of  claim 10 , wherein the imidazoquinoline compound is R848. 
     
     
         12 . The method of  claim 1 , wherein the at least one cell is a tumor cell. 
     
     
         13 . The method of  claim 1 , wherein the dendritic cells are autologous or allogeneic to the subject. 
     
     
         14 . The method of  claim 1 , wherein the dendritic cells are allogeneic cells that are HLA matched to the subject. 
     
     
         15 . The method of  claim 1 , comprising obtaining the at least one cell from a subject, wherein the cell is associated with a harmful disease state. 
     
     
         16 . The method of  claim 15 , wherein the harmful disease state is a proliferative disorder or an autoimmune disorder. 
     
     
         17 . The method of  claim 1 , wherein maturing further comprises contacting the primed dendritic cells with (i) a toll-like receptor 2 agonist, a toll-like receptor 4 agonist, or (ii) interferon gamma. 
     
     
         18 . The method of  claim 17 , wherein the toll-like receptor 2 agonist or the toll-like receptor 4 agonist is lipopolysaccharide. 
     
     
         19 . The method of  claim 1 , wherein maturing further comprises contacting the primed dendritic cells with (i) a toll-like receptor 2 agonist or a toll-like receptor 4 agonist; and (ii) interferon gamma. 
     
     
         20 . The method of  claim 1 , wherein maturing comprises contacting the primed dendritic cells with R848; lipopolysaccharide; and interferon gamma.