Optimally activated dendritic cells that induce an improved or increased anti-tumor immune response

Abstract

The present disclosure provides populations of cells comprising partially mature and optimally activated dendritic cells that can be used for administration to individuals having a cancer and/or tumor. Partially matured dendritic cells, those contacted with a dendritic cell maturation agent for about 10 to about 19 hours, upon administration efficiently take up and process tumor antigens in the area of the tumor site, complete maturation, and can subsequently migrate to the lymph nodes of a treated individual. Once in a lymph node the now fully mature antigen presenting dendritic cells secrete the appropriate cytokines (e.g., TNFα, IL-6, IL-8, and/or IL-12) and contact T cells inducing a substantial and optimal clinical and/or anti-tumor immune response.

1 . A method for treating a tumor, comprising:
 i) isolating a cell population comprising human PBMCs from peripheral blood;   ii) enriching the cell population comprising human PBMCs for human monocytic dendritic cell precursors;   iii) culturing the cell population enriched for human monocytic dendritic cell precursors with a tissue culture medium supplemented with an effective amount of a dendritic cell differentiation agent for a time period sufficient to differentiate the human monocytic dendritic cell precursors into immature human dendritic cells;   iv) culturing the cell population enriched for immature human dendritic cells with an effective amount of a dendritic cell maturation agent for about 10 to about 19 hours to partially mature and activate the immature human dendritic cells;   v) isolating and washing the partially mature and activated human dendritic cells; and   vi) administering the partially mature and activated dendritic cells directly into the tumor, into a tumor bed subsequent to surgical removal or resection of the tumor, to a tissue area surrounding the tumor, into a lymph node directly draining a tumor area, directly to a circulatory vessel duct that delivers blood or lymph to the tumor or a tumor afflicted organ, or into the circulatory system such that the cells are delivered to the tumor or tumor afflicted organ to treat said tumor.   
     
     
         2 . A method for treating a tumor, comprising:
 i) isolating a cell population comprising human monocytic dendritic cell precursors;   ii) culturing the cell population enriched for human monocytic dendritic cell precursors with a tissue culture medium supplemented with an effective amount of a dendritic cell differentiation agent for a time period sufficient to differentiate the human monocytic dendritic cell precursors into immature human dendritic cells;   iii) culturing the cell population enriched for immature human dendritic cells with an effective amount of a dendritic cell maturation agent to activate the immature human dendritic cells for about 10 to about 19 hours;   iv) isolating and washing the activated human dendritic cells; and   v) administering the activated dendritic cells directly into the tumor, into a tumor bed subsequent to surgical removal or resection of the tumor, to a tissue area surrounding the tumor, into a lymph node directly draining a tumor area, directly to a circulatory vessel duct that delivers blood or lymph to the tumor or a tumor afflicted organ, or into the circulatory system such that the cells are delivered to the tumor or tumor afflicted organ to treat said tumor.   
     
     
         3 . The method according to  claim 1 , wherein the monocytic dendritic cell precursors are obtained from skin, spleen, bone marrow, thymus, lymph nodes, umbilical cord blood, or peripheral blood. 
     
     
         4 . The method according to  claim 1 , wherein the monocytic dendritic cell precursor cells are non-activated monocytic dendritic cell precursors. 
     
     
         5 . The method according to  claim 1 , wherein the monocytic dendritic cell precursors are obtained from the individual subject to be treated. 
     
     
         6 . The method according to  claim 1 , wherein the monocytic dendritic cell precursors are obtained from a healthy individual subject HLA-matched to the individual subject to be treated. 
     
     
         7 . The method according to  claim 1 , wherein the dendritic cell differentiation agent is GM-CSF without any other cytokine, or GM-CSF in combination with IL-4, IL-7, IL-13 or IL-15. 
     
     
         8 . The method according to  claim 1 , wherein the dendritic cell maturation agent is inactivated Bacillus Calmette-Guerin (BCG), interferon γ (IFNγ), lipopolysaccharide (LPS), tumor necrosis factor α (TNFα), an imidazoquinoline compound, a synthetic double stranded polyribonucleotide, a agonist of a Toll-like receptor (TLR), a sequence of nucleic acids containing unmethylated CpG motifs known to induce the maturation of dendritic cells, or any combination thereof. 
     
     
         9 . The method according to  claim 8 , wherein the inactivated BCG comprises whole BCG, cell wall constituents of BCG, BCG-derived lipoarabidomannans, or BCG components. 
     
     
         10 . The method according to  claim 9 , wherein the inactivated BCG is heat-inactivated BCG, formalin-treated BCG, or heat-inactivated and formalin treated BCG. 
     
     
         11 . The method according to  claim 8 , wherein the effective amount of BCG is about 10 5  to 10 7  cfu per milliliter of tissue culture media and the effective amount of IFNγ is about 100 to about 1,000 Units per milliliter of tissue culture media. 
     
     
         12 . The method according to  claim 8 , wherein the imidazoquinoline compound is an imidazoquinoline-4-amine compound. 
     
     
         13 . The method according to  claim 12 , wherein the imidazoquinoline-4-amine compound is 4-amino-2-ethoxymethyl-α,α-dimethyl-1H-imidazol[4,5-c]quinolin-1-5 ethanol or 1-(2-methylpropyl)-1H-imidazo[4,5-c]quinolin-4-amine, or a derivative thereof. 
     
     
         14 . The method according to  claim 8 , wherein the synthetic double stranded polyribonucleotide is poly[I]:poly[C(12)U]. 
     
     
         15 . The method according to  claim 1 , wherein about 10 2  to about 10 10  of the partially mature and activated dendritic cells are suspended in a pharmaceutically acceptable carrier. 
     
     
         16 . The method according to  claim 1 , wherein the partially mature and activated dendritic cells are administered as an adjuvant to radiation therapy, chemotherapy, or a combination thereof. 
     
     
         17 . The method according to  claim 16 , wherein the activated dendritic cells are administered prior to, simultaneous with, or subsequent to radiation therapy, chemotherapy, or a combination thereof. 
     
     
         18 . A method for producing an anti-tumor immune response comprising administrating a composition comprising a cell population enriched for human dendritic cells that have been partially matured and optimally activated in vitro with a human dendritic cell maturation agent for about 10 to about 19 hours and a pharmaceutically acceptable carrier; wherein the composition is administered into a tumor, a tumor bed or a tissue area surrounding a tumor in an individual in need of such treatment. 
     
     
         19 . A composition comprising partially mature and optimally activated human dendritic cells induced to mature by culturing immature dendritic cells with a human dendritic cell maturation agent for about 10 to about 19 hours, wherein the partially mature and optimally activated dendritic cells produce cytokines associated with an inflammatory response. 
     
     
         20 . The composition according to  claim 19 , wherein the cytokines are tumor necrosis factor α (TNFα), interleukin 6 (IL-6), and/or interleukin 8 (IL-8).