Compositions and methods to immunize against hepatitis c virus

Abstract

Compositions comprising viral antigens and antigenic peptides corresponding to or derived from Hepatitis C virus (HCV) proteins or fragments thereof, fused to heavy and/or light chain of antibodies, or fragments thereof specific for dendritic cells (DCs) are described herein. Included herein are immunostimulatory compositions (HCV vaccines, HCV antigen presenting dendritic cells, etc.) and methods for increasing effectiveness of HCV antigen presentation by an antigen presenting cell, for a treatment, a prophylaxis or a combination thereof against hepatitis C in a human subject, and methods of providing immunostimulation by activation of one or more dendritic cells, methods to treat or prevent hepatitis C.

1 . An immunostimulatory composition for generating an immune response for a prophylaxis, a therapy, or any combination thereof against a Hepatitis C infection in a human or animal subject comprising:
 one or more antibodies or fragments thereof specific for a dendritic cell (DC); and   one or more HCV antigens attached to the one or more antibodies or fragments thereof.   
     
     
         2 . The composition of  claim 1 , further comprising at least one Toll-Like Receptor (TLR) agonist selected from the group consisting of TLR1, TLR2, TLR3, TLR4, TLR5, TLR6, TLR7, and TLR8 agonists. 
     
     
         3 . The composition of  claim 1 , further comprising an optional pharmaceutically acceptable carrier that is effective, in combination, to produce the immune response for prophylaxis, for therapy, or any combination thereof in the human or animal subject in need of immunostimulation. 
     
     
         4 . The composition of  claim 1 , wherein the antibody or fragment specific for the DC is specific for a DC specific cell surface receptor selected from an antibody that specifically binds to MHC class I, MHC class II, CD1, CD2, CD3, CD4, CD8, CD11b, CD14, CD15, CD16, CD19, CD20, CD29, CD31, CD40, CD43, CD44, CD45, CD54, CD56, CD57, CD58, CD83, CD86, CMRF-44, CMRF-56, DCIR, DC-ASPGR, CLEC-6, CD40, BDCA-2, MARCO, DEC-205, mannose receptor, Langerin, DECTIN-1, B7-1, B7-2, IFN-γ receptor and IL-2 receptor, ICAM-1, Fcγ receptor, LOX-1, and ASPGR. 
     
     
         5 . The composition of  claim 1 , wherein the HCV antigens comprise a peptide sequence derived from a HCV 1a genotype protein or a fragment thereof. 
     
     
         6 . The composition of  claim 1 , wherein the HCV antigens are selected from the group consisting of protein E1, envelope protein E2, non-structural protein NS3, non-structural protein NS4b, non-structural protein NS5b, and a fragment thereof. 
     
     
         7 . The composition of  claim 1 , wherein the one or more HCV antigens are selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14, or a fragment thereof. 
     
     
         8 . The composition of  claim 1 , wherein the composition comprises a recombinant antibody that comprises a fusion protein and the one or more HCV antigens at a C-terminal position relative to the one or more antibody or fragment thereof; a recombinant antibody or fragment thereof specific for the DC and one or more HCV antigens are fused to a C-terminus of a heavy chain of the antibody; a recombinant antibody or fragment thereof specific for the DC and one or more HCV antigens are fused to a C-terminus of a light chain of the antibody; or a recombinant antibody or fragment thereof specific for the DC and three HCV antigens or antigenic domains fused to the C-terminus of the heavy chain of the antibody or two HCV antigens or antigenic domains fused to the heavy chain of the antibody and one HCV antigen or antigenic domain fused to the light chain of the antibody. 
     
     
         9 . The composition of  claim 1 , wherein the one or more HCV antigens are selected from the group consisting of SEQ ID NO: 12-linker A-SEQ ID NO: 13, SEQ ID NO: 12-linker A-SEQ ID NO: 11, SEQ ID NO: 12-linker B-SEQ ID NO: 14, SEQ ID NO: 14-linker B-SEQ ID NO: 12, SEQ ID NO: 12-linker B-SEQ ID NO: 10, SEQ ID NO: 10-linker B-SEQ ID NO: 12, SEQ ID NO: 9-linker B-SEQ ID NO: 10, SEQ ID NO: 10-linker B-SEQ ID NO: 9, SEQ ID NO: 10-linker B-SEQ ID NO: 14, SEQ ID NO: 14-linker B-SEQ ID NO: 10, SEQ ID NO: 9-linker B-SEQ ID NO: 12, SEQ ID NO: 12-linker B-SEQ ID NO: 9, SEQ ID NO: 8-linker B-E1b. SEQ ID NO: 12-linkerB-SEQ ID NO: 10-linker C-SEQ ID NO: 14, SEQ ID NO: 12-linker B-SEQ ID NO: 14-linker C-SEQ ID NO: 10, SEQ ID NO: 10-linker B-SEQ ID NO: 12-linker C-SEQ ID NO: 14, SEQ ID NO: 10-linker B-SEQ ID NO: 14-linker C-SEQ ID NO: 12, SEQ ID NO: 14-linker B-SEQ ID NO: 12-linker C-SEQ ID NO: 10, SEQ ID NO: 14-linker B-SEQ ID NO: 10-linker C-SEQ ID NO: 12, and SEQ ID NO: 12-linker B-SEQ ID NO: 10-linker C-SEQ ID NO: 14-linker D-SEQ ID NO: 8. 
     
     
         10 . The composition of  claim 1 , wherein the one or more HCV antigens are attached to a C-terminus of a light chain of the recombinant antibody and selected from a group consisting of: SEQ ID NO: 9; SEQ ID NO: 11; or SEQ ID NO: 9 fused to the C-terminus of a light chain and SEQ ID NO: 10-linker B-SEQ ID NO: 12-linker C-SEQ ID NO: 14 fused to the C-terminus of the heavy chain of the antibody. 
     
     
         11 . The composition of  claim 1 , wherein the one or more HCV antigen are chemically coupled to the one or more antibodies or fragments thereof or are attached to the one or more antibodies or fragments thereof via an affinity association. 
     
     
         12 . The composition of  claim 1 , wherein the DC-specific antibody is humanized. 
     
     
         13 . The composition of  claim 1 , wherein the one or more HCV antigens are selected from antibodies having a heavy and a light chain combination of sequences SEQ ID NOS: 80 and 81; 82 and 83; 84 and 85; 86 and 87; 88 and 89; 90 and 91; 92 and 93; 94 and 95; 96 and 97; 98 and 99; 100 and 101; 102 and 103; 104 and 105; 106 and 107; 108 and 109; 110 and 111; 112 and 113; 114 and 115; 116 and 117; 118 and 119; 120 and 121; 122 and 123; 124 and 125; 126 and 127; 128 and 129; 130 and 131; 132 and 134; 136 and 137; 138 and 139; 140 and 141; 158 and 159; 160 and 161; 162 and 163; 164 and 165. 
     
     
         14 . A vaccine comprising:
 one or more antibodies or fragments thereof specific for a dendritic cell (DC); and   one or more HCV antigens attached to the one or more antibodies or fragments thereof and optionally a pharmaceutically acceptable carrier or an adjuvant that is effective, in combination, to produce an immune response for prophylaxis, for therapy, or any combination thereof in a subject in need of immunostimulation.   
     
     
         15 . The vaccine of  claim 14 , further comprising at least one Toll-Like Receptor (TLR) agonist selected from the group consisting of TLR1, TLR2, TLR3, TLR4, TLR5, TLR6, TLR7, and TLR8 agonists. 
     
     
         16 . The vaccine of  claim 14 , wherein the HCV antigen comprises a peptide sequence derived from a HCV 1a genotype protein or a fragment thereof, protein E1, envelope protein E2, non-structural protein NS3, non-structural protein NS4b, non-structural protein NS5b, and a fragment thereof. 
     
     
         17 . The vaccine of  claim 14 , wherein the one or more HCV antigens is selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14, or a fragment thereof. 
     
     
         18 . The vaccine of  claim 14 , wherein the composition comprises a recombinant antibody that comprises a fusion protein and the one or more HCV antigens are at a C-terminal position relative to the one or more antibody or fragment thereof within a fusion protein; one or more HCV antigens are fused to a C-terminus of a heavy chain of the antibody; one or more HCV antigens are fused to a C-terminus of a light chain of the one or more antibody or fragment thereof specific for a dendritic cell; or one or more HCV antigens or antigenic domains fused to a C-terminus of the heavy chain of the antibody and at least one HCV antigen or antigenic domain fused to the C-terminus of the light chain of the antibody. 
     
     
         19 . The vaccine of  claim 14 , wherein the vaccine comprises a recombinant antibody or fragment thereof specific for the DC and three HCV antigens or antigenic domains fused to the C-terminus of the heavy chain of the antibody or two HCV antigens or antigenic domains fused to the heavy chain of the antibody and one HCV antigen or antigenic domain fused to the light chain of the antibody. 
     
     
         20 . The vaccine of  claim 14 , wherein the one or more HCV antigens are selected from the group consisting of SEQ ID NO: 12-linker A-SEQ ID NO: 13, SEQ ID NO: 12-linker A-SEQ ID NO: 11, SEQ ID NO: 12-linker B-SEQ ID NO: 14, SEQ ID NO: 14-linker B-SEQ ID NO: 12, SEQ ID NO: 12-linker B-SEQ ID NO: 10, SEQ ID NO: 10-linker B-SEQ ID NO: 12, SEQ ID NO: 9-linker B-SEQ ID NO: 10, SEQ ID NO: 10-linker B-SEQ ID NO: 9, SEQ ID NO: 10-linker B-SEQ ID NO: 14, SEQ ID NO: 14-linker B-SEQ ID NO: 10, SEQ ID NO: 9-linker B-SEQ ID NO: 12, SEQ ID NO: 12-linker B-SEQ ID NO: 9, SEQ ID NO: 8-linker B-E1b. SEQ ID NO: 12-linkerB-SEQ ID NO: 10-linker C-SEQ ID NO: 14, SEQ ID NO: 12-linker B-SEQ ID NO: 14-linker C-SEQ ID NO: 10, SEQ ID NO: 10-linker B-SEQ ID NO: 12-linker C-SEQ ID NO: 14, SEQ ID NO: 10-linker B-SEQ ID NO: 14-linker C—SEQ ID NO: 12, SEQ ID NO: 14-linker B-SEQ ID NO: 12-linker C-SEQ ID NO: 10, SEQ ID NO: 14-linker B-SEQ ID NO: 10-linker C-SEQ ID NO: 12, and SEQ ID NO: 12-linker B-SEQ ID NO: 10-linker C-SEQ ID NO: 14-linker D-SEQ ID NO: 8. 
     
     
         21 . The vaccine of  claim 14 , wherein the one or more HCV antigens are chemically coupled to the one or more antibodies or fragments thereof or are attached to the one or more antibodies or fragments thereof via an affinity association. 
     
     
         22 . The vaccine of  claim 14 , wherein the DC-specific antibody is humanized. 
     
     
         23 . The vaccine of  claim 14 , wherein the vaccine comprises an antibody with a heavy and a light chain combination of sequences SEQ ID NOS: 80 and 81; 82 and 83; 84 and 85; 86 and 87; 88 and 89; 90 and 91; 92 and 93; 94 and 95; 96 and 97; 98 and 99; 100 and 101; 102 and 103; 104 and 105; 106 and 107; 108 and 109; 110 and 111; 112 and 113; 114 and 115; 116 and 117; 118 and 119; 120 and 121; 122 and 123; 124 and 125; 126 and 127; 128 and 129; 130 and 131; 132 and 134; 136 and 137; 138 and 139; 140 and 141; 158 and 159; 160 and 161; 162 and 163; 164 and 165. 
     
     
         24 . A Hepatitis C vaccine (HCV) comprising a fusion protein comprising:
 one or more antibodies or fragments thereof specific for a dendritic cell (DC);   one or more HCV antigens located C-terminal of the antibodies or fragments thereof;   at least one Toll-Like Receptor (TLR) agonist which is selected from the group consisting of TLR1, TLR2, TLR3, TLR4, TLR5, TLR6, TLR7, and TLR8 agonists; and   one or more optional pharmaceutically acceptable carriers and adjuvants, wherein the vaccine is effective to produce an immune response, for a prophylaxis, a therapy, or any combination thereof against hepatitis C in a human or an animal subject in need thereof.   
     
     
         25 . The vaccine of  claim 24 , wherein the vaccine comprises one or more optional agents selected from the group consisting of an agonistic anti-CD40 antibody, an agonistic anti-CD40 antibody fragment, a CD40 ligand (CD40L) polypeptide, a CD40L polypeptide fragment, anti-4-1BB antibody, an anti-4-1BB antibody fragment, 4-1BB ligand polypeptide, a 4-1BB ligand polypeptide fragment, IFN-γ, TNF-α, type 1 cytokines, type 2 cytokines or combinations and modifications thereof. 
     
     
         26 . The vaccine of  claim 24 , wherein the antibody or fragment specific for the DC is selected from an antibody that specifically binds to MHC class I, MHC class II, CD1, CD2, CD3, CD4, CD8, CD11b, CD14, CD15, CD16, CD19, CD20, CD29, CD31, CD40, CD43, CD44, CD45, CD54, CD56, CD57, CD58, CD83, CD86, CMRF-44, CMRF-56, DCIR, DC-ASPGR, CLEC-6, CD40, BDCA-2, MARCO, DEC-205, mannose receptor, Langerin, DECTIN-1, B7-1, B7-2, IFN-γ receptor and IL-2 receptor, ICAM-1, Fcγ receptor, LOX-1, and ASPGR. 
     
     
         27 . A method for increasing effectiveness of Hepatitis C virus (HCV) antigen presentation by an antigen presenting cell (APC) comprising the steps of:
 providing an antibody conjugate comprising a dendritic cell (DC) specific antibody or a fragment thereof and one or more native or engineered HCV antigenic peptides;   providing one or more APCs; and   contacting the APC with the conjugate, wherein the antibody-antigen complex is processed and presented for T cell recognition.   
     
     
         28 . The method of  claim 27 , wherein the antigen presenting cell comprises a dendritic cell (DC). 
     
     
         29 . The method of  claim 27 , wherein the DC-specific antibody or fragment is selected from an antibody that specifically binds to MHC class I, MHC class II, CD1, CD2, CD3, CD4, CD8, CD11b, CD 14, CD 15, CD 16, CD19, CD20, CD29, CD31, CD40, CD43, CD44, CD45, CD54, CD56, CD57, CD58, CD83, CD86, CMRF-44, CMRF-56, DCIR, DC-ASPGR, CLEC-6, CD40, BDCA-2, MARCO, DEC-205, mannose receptor, Langerin, DECTIN-1, B7-1, B7-2, IFN-γ receptor and IL-2 receptor, ICAM-1, Fcγ receptor, LOX-1, and ASPGR. 
     
     
         30 . The method of  claim 27 , wherein the one or more native or engineered HCV antigenic peptide is selected from the group consisting of: SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 6, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14, E1b, and a fragment thereof. 
     
     
         31 . The method of  claim 27 , wherein the DC-specific antibody is humanized. 
     
     
         32 . A method for increasing effectiveness of antigen presentation by an antigen presenting cell (APC) comprising the steps of:
 isolating and purifying one or more dendritic cell (DC)-specific antibodies or a fragments thereof;   providing one or more HCV antigens or antigenic peptides;   loading or chemically coupling the one or more HCV antigens or antigenic peptides to the DC-specific antibody to form an antibody-antigen conjugate; and   contacting the antigen presenting cell with the conjugate, wherein the antibody-antigen complex is processed and presented for T cell recognition.   
     
     
         33 . The method of  claim 32 , further comprising the optional steps of:
 adding one or more optional agents selected from the group consisting of an agonistic anti-CD40 antibody, an agonistic anti-CD40 antibody fragment, a CD40 ligand (CD40L) polypeptide, a CD40L polypeptide fragment, anti-4-1BB antibody, an anti-4-1BB antibody fragment, 4-1BB ligand polypeptide, a 4-1BB ligand polypeptide fragment, IFN-γ, TNF-α, type 1 cytokines, type 2 cytokines or combinations and modifications thereof to the antibody-antigen conjugate and the TLR agonist prior to contacting the antigen presenting cells; and   measuring a level of one or more agents selected from the group consisting of IFN-γ, TNF-α, IL-12p40, IL-4, IL-5, and IL-13, wherein a change in the level of the one or more agents is indicative of the increase in the effectiveness antigen presentation by the antigen presenting cell.   
     
     
         34 . The method of  claim 32 , wherein the APC comprises a dendritic cell (DC). 
     
     
         35 . The method of  claim 32 , wherein the DC-specific antibody or fragment is selected from an antibody that specifically binds to MHC class I, MHC class II, CD1, CD2, CD3, CD4, CD8, CD11b, CD14, CD 15, CD16, CD19, CD20, CD29, CD31, CD40, CD43, CD44, CD45, CD54, CD56, CD57, CD58, CD83, CD86, CMRF-44, CMRF-56, DCIR, DC-ASPGR, CLEC-6, CD40, BDCA-2, MARCO, DEC-205, mannose receptor, Langerin, DECTIN-1, B7-1, B7-2, IFN-γ receptor and IL-2 receptor, ICAM-1, Fcγ receptor, LOX-1, and ASPGR. 
     
     
         36 . The method of  claim 32 , wherein the HCV antigen is selected from the group consisting of protein E1, envelope protein E2, non-structural protein NS3, non-structural protein NS4b, non-structural protein NS5b, and a fragment thereof. 
     
     
         37 . The method of  claim 32 , wherein the one or more HCV antigens is selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, and a fragment thereof. 
     
     
         38 . The method of  claim 32 , wherein the DC-specific antibody is humanized. 
     
     
         39 . The method of  claim 32 , further comprising the optional step of adding one or more optional agents selected from the group consisting of an agonistic anti-CD40 antibody, an agonistic anti-CD40 antibody fragment, a CD40 ligand (CD40L) polypeptide, a CD40L polypeptide fragment, anti-4-1BB antibody, an anti-4-1BB antibody fragment, 4-1BB ligand polypeptide, a 4-1BB ligand polypeptide fragment, IFN-γ, TNF-α, type 1 cytokines, type 2 cytokines or combinations and modifications thereof. 
     
     
         40 . The method of  claim 32 , further comprising adding at least one Toll-Like Receptor (TLR) agonist which is selected from the group consisting of TLR1, TLR2, TLR3, TLR4, TLR5, TLR6, TLR7, and TLR8 agonists. 
     
     
         41 . A Hepatitis C virus antigen presenting dendritic cell (DC) comprising one or more isolated dendritic cells (DCs) in contact with a fusion protein comprising an antibody or fragment thereof specific for the DC, the fusion protein further comprising a HCV peptide. 
     
     
         42 . A method for generating a Hepatitis C virus (HCV) presenting dendritic cells (DCs) in a human subject comprising the steps of
 providing one or more DCs;   incubating the dendritic cells with a fusion protein, wherein the fusion protein comprises an antibody or fragment thereof specific for a dendritic cell and a HCV antigen fused to the antibody or fragment thereof.   
     
     
         43 . The method of  claim 42 , further comprising administering to the subject an effective amount of IFNA, Ribavirin, or a combination thereof. 
     
     
         44 . A vaccine comprising one or more antibodies or fragments thereof specific for a dendritic cell (DC) and one or more HCV antigens or antigenic domains attached to the one or more antibodies or fragments thereof, wherein the vaccine has a general structure given by:
 H-w,   H-w-x,   H-w-x-y, or   H-w-x-y-z,   wherein H represents a heavy chain of an antibody or a fragment thereof specific for a DC, w, x, y, and z represent one or more HCV antigens or domains selected from the group consisting of protein E1, envelope protein E2, non-structural protein NS3, non-structural protein NS4b, non-structural protein NS5b, or any combinations thereof.   
     
     
         45 . The vaccine of  claim 44 , wherein w comprises the HCV antigenic domains selected from the group consisting of ProtA, ProtB, HelB, Palm, E1b, and E2. 
     
     
         46 . The vaccine of  claim 44 , wherein x comprises the HCV antigenic domains selected from the group consisting of HelC, HelA, Palm, ProtA, ProtB, and E1b. 
     
     
         47 . The vaccine of  claim 44 , wherein y comprises the HCV antigenic domains selected from the group consisting of Palm, ProtB, and Protb. 
     
     
         48 . The vaccine of  claim 44 , wherein z comprises HCV antigenic domains selected from E2, ProtA, and HelB. 
     
     
         49 . The vaccine of  claim 44 , wherein the one or more HCV antigens or antigenic domains are linked or attached to one another by one or more flexible linkers. 
     
     
         50 . The vaccine of  claim 44 , wherein the vaccine comprises H-ProtA, H-ProtB, H-HelB, H-Palm, H-E1b, H-E2, H-HelB-HelC, H-HelB-HelA, H-HelB-Palm, H-Palm-HelB, H-HelB-ProtB, H-ProtB-HelB, H-ProtA-ProtB, H-ProtB-ProtA, H-ProtB-Palm, H-Palm-ProtB, H-ProtA-HelB, H-HelB-ProtA, H-E2-E1b, H-HelB-ProtB-Palm, H-HelB-Palm-ProtB, H-ProtB-HelB-Palm, H-ProtB-Palm-HelB, H-Palm-HelB-ProtB, H-Palm-ProtB-HelB, H-HelB-ProtB-Palm-E2, or any combinations thereof. 
     
     
         51 . A vaccine comprising one or more antibodies or fragments thereof specific for a dendritic cell (DC) and one or more HCV antigens or antigenic domains attached to the one or more antibodies or fragments thereof, wherein the vaccine has a general structure given by L-w-x-y-z, wherein L represents a light chain of an antibody or a fragment thereof specific for a DC, w, x, y, and z represent one or more HCV antigens or domains selected from the group consisting of protein E1, envelope protein E2, non-structural protein NS3, non-structural protein NS4b, non-structural protein NS5b, or any combinations thereof. 
     
     
         52 . The composition of  claim 51 , wherein w, x, y, and z comprise HCV antigenic domains selected from the group consisting of ProtA, ProtB, HelB, HelA, HelC, Palm, E1a, E1b, and E2. 
     
     
         53 . The composition of  claim 51 , wherein the vaccine comprises L-ProtA-HelA-E1b-HelB. 
     
     
         54 . A vaccine comprising one or more antibodies or fragments thereof specific for a dendritic cell (DC) and one or more HCV antigens or antigenic domains attached to the one or more antibodies or fragments thereof, wherein the vaccine has a general structure given by: 
       
         
           
           
               
               
           
         
         wherein H represents a heavy chain of an antibody or a fragment thereof specific for a DC, L represents a light chain of an antibody or a fragment thereof specific for the DC, w, x, y, and z represent one or more HCV antigens or domains selected from the group consisting of protein E1, envelope protein E2, non-structural protein NS3, non-structural protein NS4b, non-structural protein NS5b, or any combinations thereof. 
       
     
     
         55 . The composition of  claim 54 , wherein w, x, y, and z comprise HCV antigenic domains selected from the group consisting of ProtA, ProtB, HelB, HelA, HelC, Palm, E1a, E1b, and E2. 
     
     
         56 . The composition of  claim 54 , wherein the vaccine comprises 
       
         
           
           
               
               
           
         
       
     
     
         57 . The composition of  claim 54 , wherein the vaccine comprises