Use of gilz protein expressed in dendritic cells to modulate an antigen-specific immune response

Abstract

A pharmaceutical composition comprising at least dendritic cells modified with a molecule selected from the group consisting of: a GILZ protein or a functional fragment of at least 5 consecutive amino acids of said protein; a small interfering RNA (siRNA) targeting the GILZ transcript or an antisense oligodeoxynucleotide complementary to said transcript; and a recombinant expression vector containing a polynucleotide encoding said GILZ protein, said fragment, said siRNA or said antisense oligodeoxynucleotide complementary to said transcript and a pharmaceutically acceptable carrier. In addition, a method for treatment of autoimmune diseases, inflammatory diseases, allergies, transplant rejection and graft-versus-host disease, cancers or pathogenic microorganism infections by administering an effective amount of said pharmaceutical composition to a subject in need thereof.

1 - 16 . (canceled) 
     
     
         17 . A pharmaceutical composition comprising at least dendritic cells modified with a molecule selected from the group consisting of: a GILZ protein or a functional fragment of at least 5 consecutive amino acids of said protein, a small interfering RNA (siRNA) targeting the GILZ transcript or an antisense oligodeoxynucleotide complementary to said transcript, and a recombinant expression vector comprising a polynucleotide encoding said GILZ protein, said fragment, said siRNA or said antisense oligodeoxynucleotide complementary to said transcript and a pharmaceutically acceptable carrier. 
     
     
         18 . The pharmaceutical composition as claimed in  claim 17 , further comprising an antigen. 
     
     
         19 . The pharmaceutical composition as claimed in  claim 18 , wherein said antigen is loaded onto said modified dendritic cells or is presented by said cells. 
     
     
         20 . A method for treatment of autoimmune diseases, inflammatory diseases, allergies, transplant rejection and graft-versus-host disease, cancers or pathogenic microorganism infections comprising administering an effective amount of the composition of  claim 17  to a subject in need thereof. 
     
     
         21 . A pharmaceutical composition comprising at least: a) a GILZ protein, a functional fragment of at least 5 consecutive amino acids of said protein, a small interfering RNA (siRNA) targeting the GILZ transcript, an antisense oligodeoxynucleotide complementary to said transcript or a recombinant expression vector comprising a polynucleotide encoding said GILZ protein, said fragment, said siRNA or said antisense oligodeoxynucleotide complementary to said transcript, and b) an antigen of interest and/or a molecule for targeting and/or crossing the plasma membrane of dendritic cells, and a pharmaceutically acceptable carrier. 
     
     
         22 . The pharmaceutical composition as claimed in  claim 21 , wherein said composition comprises a chimeric GILZ protein or peptide comprising the sequence of a peptide for targeting and/or crossing the plasma membrane of dendritic cells and/or the sequence of said antigen of interest defined in b). 
     
     
         23 . The pharmaceutical composition as claimed in  claim 21 , wherein said composition comprises one or more recombinant expression vectors encoding: said GILZ protein, said fragment, said siRNA or said antisense oligodeoxynucleotide complementary to said transcript, defined in a), and said antigen and/or said molecule defined in b). 
     
     
         24 . The pharmaceutical composition as claimed in  claim 21 , wherein said composition comprises a recombinant expression vector for a chimeric GILZ protein or peptide comprising the sequence of a peptide for targeting and/or crossing the plasma membrane of dendritic cells and/or the sequence of said antigen of interest defined in b). 
     
     
         25 . (canceled) 
     
     
         26 . A method for inhibiting suppressor/regulatory T lymphocytes in vitro, using a tissue sample containing CD4 +  and/or CD8 +  T cells, comprising the steps of:
 a2) preparing autologous dendritic cells in which a GILZ protein is inhibited, and, simultaneously or sequentially, 
 b2) incubating said dendritic cells obtained in a2) with an antigen and said tissue sample containing CD4 +  and/or CD8 +  T cells. 
 
     
     
         27 . A pharmaceutical composition comprising at least: a) a GILZ protein, a functional fragment of at least 5 consecutive amino acids of said protein, a GILZ modulator, or a recombinant expression vector comprising a polynucleotide encoding said GILZ protein, said fragment, or said modulator, the recombinant vector being isolated or expressed in modified dendric cells, and b) an antigen of interest and/or a molecule for targeting and/or crossing the plasma membrane of dendritic cells, and a pharmaceutically acceptable carrier. 
     
     
         28 . The pharmaceutical composition of  claim 27 , wherein the antigen is selected from  Phleum pratense, Dermatophagoides pteronyssinus  and farinae, a latex antigen such as Hev b 1 to 11, proinsulin, MAG, thyroperoxidase and thyreoglobulin. 
     
     
         29 . The pharmaceutical composition of  claim 27 , wherein a) is a GILZ inhibitor or a recombinant expression vector comprising the inhibitor, the vector being isolated or expressed in modified dendritic cells, and said antigen defined in b) is an antigen of vaccine of interest. 
     
     
         30 . The pharmaceutical composition of  claim 29 , wherein said inhibitor is a small interfering RNA that targets a GILZ transcript or an antisense oligodeoxynucleotide complementary to said transcript. 
     
     
         31 . The pharmaceutical composition of  claim 29 , wherein said inhibitor is a GILZ antagonist selected from the group consisting of: an antagonist of the akt kinase and an activator of said kinase. 
     
     
         32 . The pharmaceutical composition of  claim 27 , wherein a) is a GILZ protein, a functional fragment of at least 5 consecutive amino acids of said protein, a GILZ activator, or a recombinant expression vector encoding said GILZ protein, said fragment, or said activator, the vector being isolated or expressed in modified dendritic cells, and said antigen defined in b) is selected from the group consisting of: an autoantigen, an allergen, an antigen involved in a chronic inflammatory disease and a transplantation antigen. 
     
     
         33 . The pharmaceutical composition of  claim 32 , wherein said GILZ protein is the human protein. 
     
     
         34 . The pharmaceutical composition of  claim 32 , wherein said activator is an inducer of GILZ gene expression, selected from the group consisting of: dexamethasone, IL-10 and TGFβ. 
     
     
         35 . The pharmaceutical composition of  claim 32 , wherein said antigen in b) is selected from  Phleum pratense, Dermatophagoides pteronyssinus  and farinae, a latex antigen such as Hev b 1 to 11, proinsulin, MAG, thyroperoxidase and thyreoglobulin. 
     
     
         36 . The pharmaceutical composition of  claim 27 , wherein said molecule for targeting dendritic cells is a ligand of a membrane antigen or receptor selected from the group consisting of: DC-SIGN, CD40, DEC-205, langerin, the mannose receptor and a scavenger receptor. 
     
     
         37 . The pharmaceutical composition of  claim 27 , wherein said molecule for targeting dendritic cells is an antibody directed against a membrane antigen or receptor selected from the group consisting of: DC-SIGN, CD40, DEC-205, langerin, the mannose receptor and a scavenger receptor. 
     
     
         38 . The pharmaceutical composition of  claim 27 , wherein said vector comprises an expression cassette that includes a promoter for a gene expressed specifically or preferentially in dendritic cells. 
     
     
         39 . The pharmaceutical composition of  claim 38 , wherein said promoter is the promoter of a gene encoding a protein selected from the group consisting of: DC-SIGN, CD11c, a molecule of the major histocompatibility complex and langerin. 
     
     
         40 . The pharmaceutical composition of  claim 27 , wherein said expression vector defined in a) is a lentivirus. 
     
     
         41 . The pharmaceutical composition of  claim 27 , wherein said GILZ protein or said fragment are in the form of a chimeric protein or peptide that comprises the sequence of a peptide for targeting and/or crossing the plasma membrane of dendritic cells and/or the sequence of said antigen of interest defined in b). 
     
     
         42 . The pharmaceutical composition of  claim 27 , wherein said recombinant expression vector defined in a) encodes a chimeric GILZ protein or a chimeric GILZ fragment comprising the sequence of a peptide for targeting and/or crossing the plasma membrane of dendritic cells and/or the sequence of said antigen of interest defined in b). 
     
     
         43 . The pharmaceutical composition of  claim 27 , wherein the antigen is loaded onto said modified dendritic cells or is presented by said cells. 
     
     
         44 . A method for treatment of autoimmune diseases, inflammatory diseases, allergies, transplant rejection and graft-versus-host disease, cancers or pathogenic microorganism infections comprising administering an effective amount of the composition of  claim 27  to a subject in need thereof. 
     
     
         45 . A method for treatment of autoimmune diseases, inflammatory diseases, allergies, transplant rejection and graft-versus-host disease, cancers or pathogenic microorganism infections comprising administering an effective amount of the composition of  claim 21  to a subject in need thereof. 
     
     
         46 . The pharmaceutical composition of  claim 29 , wherein the antigen is a tumor antigen or an antigen of a pathogenic microorganism. 
     
     
         47 . The pharmaceutical composition of  claim 31 , wherein the activator of said kinase is a chemokine RANTES.