Method for generating highly active human dendritic cells from monocytes

Abstract

The present invention relates to a process for deriving dendritic cells from mononuclear cells in culture comprising the step of putting in contact type I IFN with said mononuclear cells. Dendritic cells suitable as cellular adjuvants in prophylactic as well as therapeutic vaccination of animal and human beings, are obtainable thereby, after a single step treatment in a brief period of time. Dendritic cells obtainable thereby, pharmaceutical compositions including them, in particular a vaccine comprising said cells as active principle, and a method of treatment of a pathology associated with the presence of an antigen in human beings, are further objects of the invention, as well as a kit for deriving said dendritic cells and a method for the ex vivo expansion of T cells using them.

1 . A process for the preparation of dendritic cells comprising the step of culturing mononuclear cells in a culture medium containing type I interferon, wherein said mononuclear cells are chosen from the group consisting of total peripheral blood mononuclear cells, adherent peripheral blood mononuclear cells and highly purified CD14 +  monocytes isolated from peripheral blood mononuclear cells.  
     
     
         2 . A process according to  claim 1 , wherein said dendritic cells are obtained in no more than three days.  
     
     
         3 . A process according to  claim 1 , wherein said type I interferon is chosen from the group consisting of natural or recombinant IFN□, natural or recombinant INFO consensus interferon and any synthetic type I interferon.  
     
     
         4 . A process according to  claim 1 , wherein the concentration of said type I interferon in the culture medium is greater than 100 IU/ml.  
     
     
         5 . A process according to  claim 4  wherein said concentration is between 100 and 10.000 IU/ml.  
     
     
         6 . A process according to  claim 5  wherein said concentration is between 400 and 10.000 IU/ml.  
     
     
         7 . A process according to  claim 6  wherein said concentration is between 500 and 2.000 IU/ml.  
     
     
         8 . A process according to  claim 7  wherein said concentration is about 1000 IU/ml.  
     
     
         9 . A process according to  claim 1  wherein said culture medium also contains a cell growth factor.  
     
     
         10 . A process according to  claim 9  wherein said cell growth factor is GM-CSF.  
     
     
         11 . A process according to  claim 10  wherein said the concentration of said GM-CSF in the medium is between 250 and 1000 IU/ml.  
     
     
         12 . A process according to  claim 1  wherein said process further comprises the step of contacting the dendritic cells obtained with a maturation agent.  
     
     
         13 . Dendritic cells obtainable with a process according to  claim 1 .  
     
     
         14 . Dendritic cells according to  claim 13  said cells having been loaded with antigenic peptides or proteins, with a cellular extract containing at least one antigen or with nucleic acid molecules encoding for antigens to which an immune response is of interest  
     
     
         15 . Dendritic cells according to  claim 13  wherein said cells are in a dehydrated or frozen form in an appropriate cryo-preservative medium.  
     
     
         16 . A kit for preparing dendritic cells according to  claim 13  comprising: 
 a) single use elements necessary for the culture and the washing of the cells;  
 b) a composition comprising type I IFN and compatible additives;  
 c) optionally a composition comprising a cell growth factor and compatible additives; and  
 d) optionally a composition comprising antigens or nucleic acids encoding for antigens to which an immune response is of interest.  
 
     
     
         17 . A pharmaceutical composition or a vaccine comprising, as an adjuvant, the dendritic cells according to  claim 13  together with at least one immunogen and a pharmaceutically acceptable vehicle or an auxiliary agent.  
     
     
         18 . A pharmaceutical composition or a vaccine comprising, as an active principle, the dendritic cells according to  claim 13  together with a pharmaceutically acceptable vehicle or auxiliary agent.  
     
     
         19 . (cancelled)  
     
     
         20 . Use of dendritic cells according to  claim 13  for the preparation of a vaccine or a pharmaceutical composition for the prevention or the treatment of a pathology associated with the presence of an antigen in the human body.  
     
     
         21 . Use according to  claim 20  wherein said pathology is an infectious or neoplastic disease.  
     
     
         22 . Use according to  claim 21  wherein said infectious disease is a viral infection.  
     
     
         23 . Use according to  claim 22  wherein said viral infection is a HIV, a HBV or a HCV infection.  
     
     
         24 . Use according to  claim 20  wherein said neoplastic disease is a lymphoma.  
     
     
         25 . Use according to  claim 21  wherein said neoplastic disease is virally induced.  
     
     
         26 . Use according to  claim 25  wherein said neoplastic disease is induced by Epstein-Barr virus.  
     
     
         27 . Use according to  claim 20  wherein said pharmaceutical composition is suitable for administration at the site of infection or within the tumour.  
     
     
         28 . A method for the ex-vivo expansion of T cells, comprising the step of putting in contact said T cells with the dendritic cells according to  claim 13 .  
     
     
         29 . (cancelled)  
     
     
         30 . A pharmaceutical composition containing, as active principle, the T cells according to  claim 28.