Compositions for the preparation of mature dendritic cells

Abstract

The invention relates to a method for in vitro maturation of at least one immate dendritic cell, comprising stimulating said immature dendritic cell with TNFα, IL-1β, IFNγ, a TLR7/8 agonist and prostaglandin E2(PG). Furthermore, the invention elates to a composition comprising these factors as well as to mature dendritic cells produced by a method of the invention.

1 - 22 . (canceled) 
     
     
         23 . A mature dendritic cell or a population of mature dendritic cells, obtainable by a method for in vitro maturation of at least one immature dendritic cell, comprising stimulating said immature dendritic cell with TNFα, IL-1β, IFNγ, a TLR7/8 agonist and prostaglandin E2 (PG2). 
     
     
         24 . The mature dendritic cell or a population of mature dendritic cells of  claim 23 , wherein the method further comprises stimulating said immature dendritic cell with a TLR3 agonist. 
     
     
         25 . The mature dendritic cell or a population of mature dendritic cells of  claim 24 , wherein the TLR3 agonist is polyI:C. 
     
     
         26 . The mature dendritic cell or a population of mature dendritic cells of  claim 23 , wherein said substances are part of a composition added to the culture medium of said immature dendritic cell. 
     
     
         27 . The mature dendritic cell or a population of mature dendritic cells of  claim 23 , wherein said TLR7/8 agonist is an imidazoquinilone type immune response modifying compound. 
     
     
         28 . The mature dendritic cell or a population of mature dendritic cells of  claim 27 , wherein said imidazoquinilone type immune response modifying compound is 4-amino-2-ethoxymethyl-α,α-dimethyl-1H-imidazol[4,5-c]quinoline-1-ethanol (R848). 
     
     
         29 . The mature dendritic cell or a population of mature dendritic cells of  claim 23 , wherein said immature dendritic cell is a monocyte derived immature dendritic cell, or wherein said immature dendritic cell is obtained directly from peripheral blood. 
     
     
         30 . The mature dendritic cell or a population of mature dendritic cells of  claim 29 , wherein said immature dendritic cell is derived from human peripheral blood mononuclear cells, monocytes, other myeloid progenitor cells, or from CD34 positive progenitor cells by in vitro differentiation to CD14 positive cells. 
     
     
         31 . The mature dendritic cell or a population of mature dendritic cells of  claim 23 , wherein the immature dendritic cell is obtained by incubating human peripheral blood mononuclear cells, monocytes or other myeloid progenitor cells with GM-CSF and IL-4 or IL-13. 
     
     
         32 . The mature dendritic cell or a population of mature dendritic cells of  claim 23 , wherein the immature dendritic cell is of human origin. 
     
     
         33 . The mature dendritic cell or a population of mature dendritic cells of  claim 23 , wherein the method comprises the following steps:
 a) preparing mononuclear cells from peripheral blood, preferably wherein said mononuclear cells are obtained by leukapheresis from peripheral blood,   b) incubating the mononuclear cells of step a) with GM-CSF and IL-4 or IL-13,   c) incubating the cells obtained in step b) with a cocktail comprising TNFα, IL-1β, IFNγ, a TLR7/8 agonist, prostaglandin E2 (PG), and, optionally, a TLR3 agonist, preferably poly I:C, and   d) harvesting the mature dendritic cell or cells.   
     
     
         34 . The mature dendritic cell or a population of mature dendritic cells of  claim 33 , wherein in step a) the mononuclear cells are obtained by leukapheresis from peripheral blood,
 wherein said incubation in step b) takes 1 to 9 days, 2 to 9 days, or 2 to 6 days, or wherein said incubation in step c) takes 12 h to 72 h, 20 h or 24 h.   
     
     
         35 . The mature dendritic cell or a population of mature dendritic cells of  claim 23 , wherein the mature dendritic cell or cells is/are further loaded in vitro with one or more antigens. 
     
     
         36 . The mature dendritic cell or a population of mature dendritic cells of  claim 35 , wherein said antigen or antigens are supposed to trigger the effector T cell maturation within the lymph nodes. 
     
     
         37 . The mature dendritic cell or a population of mature dendritic cells of  claim 35 , wherein said loading is performed by incubating the mature dendritic cell or cells with at least one peptide of said antigen or by transfecting the dendritic cell or cells with antigen encoding RNA or DNA. 
     
     
         38 . A pharmaceutical composition comprising the mature dendritic cell or the population of mature dendritic cells of  claim 23 . 
     
     
         39 . A method of treating a disease selected from the group consisting of tumorigenic and infectious diseases in a patient in need of such treatment, comprising administering to said patient a therapeutically effective dose of the mature dendritic cell or the population of mature dendritic cells of  claim 23 . 
     
     
         40 . A method for the preparation of the mature dendritic cell or the population of mature dendritic cells of  claim 23  comprising the combined use of TNFα, IL-1β, IFNγ, a TLR7/8 agonist, prostaglandin E2 (PG2) and, optionally, a TLR3 agonist, preferably poly I:C. 
     
     
         41 . The method of  claim 40 , wherein said TLR7/8 agonist is an imidazoquinilone type immune response modifying compound. 
     
     
         42 . The method of  claim 40 , wherein said imidazoquinilone type immune response modifying compound is 4-amino-2-ethoxymethyl-α,α-dimethyl-1H-imidazol[4,5-c]quinoline-1-ethanol (R848).