JP2012507280A - 改良型hcvワクチンおよびその使用方法 - Google Patents
改良型hcvワクチンおよびその使用方法 Download PDFInfo
- Publication number
- JP2012507280A JP2012507280A JP2011534469A JP2011534469A JP2012507280A JP 2012507280 A JP2012507280 A JP 2012507280A JP 2011534469 A JP2011534469 A JP 2011534469A JP 2011534469 A JP2011534469 A JP 2011534469A JP 2012507280 A JP2012507280 A JP 2012507280A
- Authority
- JP
- Japan
- Prior art keywords
- seq
- nucleic acid
- protein
- vaccine
- hcv
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 238000000034 method Methods 0.000 title claims abstract description 64
- 229960005486 vaccine Drugs 0.000 title claims description 50
- 230000001976 improved effect Effects 0.000 title abstract description 18
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 117
- 102000004169 proteins and genes Human genes 0.000 claims abstract description 93
- 150000007523 nucleic acids Chemical class 0.000 claims abstract description 83
- 102000039446 nucleic acids Human genes 0.000 claims abstract description 79
- 108020004707 nucleic acids Proteins 0.000 claims abstract description 78
- 230000028993 immune response Effects 0.000 claims abstract description 42
- 108010008038 Synthetic Vaccines Proteins 0.000 claims abstract description 32
- 229940124551 recombinant vaccine Drugs 0.000 claims abstract description 32
- 230000001939 inductive effect Effects 0.000 claims abstract description 26
- 239000008194 pharmaceutical composition Substances 0.000 claims abstract description 25
- 229940124590 live attenuated vaccine Drugs 0.000 claims abstract description 8
- 229940023012 live-attenuated vaccine Drugs 0.000 claims abstract description 8
- 125000003275 alpha amino acid group Chemical group 0.000 claims abstract 4
- 239000002773 nucleotide Substances 0.000 claims description 92
- 125000003729 nucleotide group Chemical group 0.000 claims description 92
- 239000012634 fragment Substances 0.000 claims description 41
- FWMNVWWHGCHHJJ-SKKKGAJSSA-N 4-amino-1-[(2r)-6-amino-2-[[(2r)-2-[[(2r)-2-[[(2r)-2-amino-3-phenylpropanoyl]amino]-3-phenylpropanoyl]amino]-4-methylpentanoyl]amino]hexanoyl]piperidine-4-carboxylic acid Chemical compound C([C@H](C(=O)N[C@H](CC(C)C)C(=O)N[C@H](CCCCN)C(=O)N1CCC(N)(CC1)C(O)=O)NC(=O)[C@H](N)CC=1C=CC=CC=1)C1=CC=CC=C1 FWMNVWWHGCHHJJ-SKKKGAJSSA-N 0.000 claims description 39
- 238000004520 electroporation Methods 0.000 claims description 36
- 108020004414 DNA Proteins 0.000 claims description 32
- 208000015181 infectious disease Diseases 0.000 claims description 23
- 239000013612 plasmid Substances 0.000 claims description 22
- 239000000203 mixture Substances 0.000 claims description 20
- 230000002238 attenuated effect Effects 0.000 claims description 13
- 102000053602 DNA Human genes 0.000 claims description 5
- 206010046865 Vaccinia virus infection Diseases 0.000 claims description 5
- 208000007089 vaccinia Diseases 0.000 claims description 5
- 101800001020 Non-structural protein 4A Proteins 0.000 abstract description 79
- 230000002163 immunogen Effects 0.000 abstract description 39
- 241000711549 Hepacivirus C Species 0.000 description 89
- 150000001413 amino acids Chemical group 0.000 description 83
- 229940024606 amino acid Drugs 0.000 description 67
- 101710144111 Non-structural protein 3 Proteins 0.000 description 44
- 210000004027 cell Anatomy 0.000 description 39
- 229940021995 DNA vaccine Drugs 0.000 description 35
- 108010041986 DNA Vaccines Proteins 0.000 description 34
- 230000003053 immunization Effects 0.000 description 30
- 238000002649 immunization Methods 0.000 description 27
- 230000014509 gene expression Effects 0.000 description 26
- 230000024932 T cell mediated immunity Effects 0.000 description 20
- 241000699670 Mus sp. Species 0.000 description 18
- 241000700605 Viruses Species 0.000 description 18
- 230000002068 genetic effect Effects 0.000 description 18
- 108090000765 processed proteins & peptides Proteins 0.000 description 18
- 241000282560 Macaca mulatta Species 0.000 description 17
- 108010074328 Interferon-gamma Proteins 0.000 description 15
- 108091035707 Consensus sequence Proteins 0.000 description 14
- 230000004044 response Effects 0.000 description 14
- 102100037850 Interferon gamma Human genes 0.000 description 13
- 230000005867 T cell response Effects 0.000 description 13
- 239000003623 enhancer Substances 0.000 description 12
- 108091026890 Coding region Proteins 0.000 description 11
- 101710174009 Suppressor of RNA silencing p3 Proteins 0.000 description 11
- 244000052769 pathogen Species 0.000 description 11
- 210000004988 splenocyte Anatomy 0.000 description 11
- 108020004705 Codon Proteins 0.000 description 10
- 238000011510 Elispot assay Methods 0.000 description 10
- 241001465754 Metazoa Species 0.000 description 10
- -1 TNFβ Proteins 0.000 description 10
- 238000003114 enzyme-linked immunosorbent spot assay Methods 0.000 description 10
- 229940031626 subunit vaccine Drugs 0.000 description 10
- 230000005847 immunogenicity Effects 0.000 description 9
- 238000005457 optimization Methods 0.000 description 9
- 230000008488 polyadenylation Effects 0.000 description 9
- 230000001105 regulatory effect Effects 0.000 description 9
- 239000013598 vector Substances 0.000 description 9
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 8
- 229940031551 inactivated vaccine Drugs 0.000 description 8
- 241000282693 Cercopithecidae Species 0.000 description 7
- 230000001154 acute effect Effects 0.000 description 7
- 239000013604 expression vector Substances 0.000 description 7
- 230000036039 immunity Effects 0.000 description 7
- 230000014616 translation Effects 0.000 description 7
- 241000701022 Cytomegalovirus Species 0.000 description 6
- HVLSXIKZNLPZJJ-TXZCQADKSA-N HA peptide Chemical group C([C@@H](C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](C(C)C)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)N[C@@H](C)C(O)=O)NC(=O)[C@H]1N(CCC1)C(=O)[C@@H](N)CC=1C=CC(O)=CC=1)C1=CC=C(O)C=C1 HVLSXIKZNLPZJJ-TXZCQADKSA-N 0.000 description 6
- 101001109503 Homo sapiens NKG2-C type II integral membrane protein Proteins 0.000 description 6
- 241000829100 Macaca mulatta polyomavirus 1 Species 0.000 description 6
- 102100022683 NKG2-C type II integral membrane protein Human genes 0.000 description 6
- 229940031567 attenuated vaccine Drugs 0.000 description 6
- 230000008713 feedback mechanism Effects 0.000 description 6
- 230000006870 function Effects 0.000 description 6
- 239000007927 intramuscular injection Substances 0.000 description 6
- 238000010255 intramuscular injection Methods 0.000 description 6
- 210000003819 peripheral blood mononuclear cell Anatomy 0.000 description 6
- 230000003389 potentiating effect Effects 0.000 description 6
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 6
- 102000004196 processed proteins & peptides Human genes 0.000 description 6
- 210000001519 tissue Anatomy 0.000 description 6
- 102100022203 Tumor necrosis factor receptor superfamily member 25 Human genes 0.000 description 5
- 210000004899 c-terminal region Anatomy 0.000 description 5
- 239000003795 chemical substances by application Substances 0.000 description 5
- 238000013461 design Methods 0.000 description 5
- 208000006454 hepatitis Diseases 0.000 description 5
- 230000010354 integration Effects 0.000 description 5
- 230000003834 intracellular effect Effects 0.000 description 5
- 239000000523 sample Substances 0.000 description 5
- 208000005176 Hepatitis C Diseases 0.000 description 4
- 241000701044 Human gammaherpesvirus 4 Species 0.000 description 4
- 241000699666 Mus <mouse, genus> Species 0.000 description 4
- 108091028043 Nucleic acid sequence Proteins 0.000 description 4
- 108010038512 Platelet-Derived Growth Factor Proteins 0.000 description 4
- 102000010780 Platelet-Derived Growth Factor Human genes 0.000 description 4
- 108010076504 Protein Sorting Signals Proteins 0.000 description 4
- 102000002067 Protein Subunits Human genes 0.000 description 4
- 108010001267 Protein Subunits Proteins 0.000 description 4
- 101710172711 Structural protein Proteins 0.000 description 4
- 208000037621 acute hepatitis C virus infection Diseases 0.000 description 4
- 238000010171 animal model Methods 0.000 description 4
- 239000000427 antigen Substances 0.000 description 4
- 108091007433 antigens Proteins 0.000 description 4
- 102000036639 antigens Human genes 0.000 description 4
- 230000001684 chronic effect Effects 0.000 description 4
- CVSVTCORWBXHQV-UHFFFAOYSA-N creatine Chemical compound NC(=[NH2+])N(C)CC([O-])=O CVSVTCORWBXHQV-UHFFFAOYSA-N 0.000 description 4
- 230000000694 effects Effects 0.000 description 4
- 230000001965 increasing effect Effects 0.000 description 4
- 239000007924 injection Substances 0.000 description 4
- 238000002347 injection Methods 0.000 description 4
- 210000004185 liver Anatomy 0.000 description 4
- 238000013507 mapping Methods 0.000 description 4
- 230000004048 modification Effects 0.000 description 4
- 238000012986 modification Methods 0.000 description 4
- 210000003205 muscle Anatomy 0.000 description 4
- 230000007935 neutral effect Effects 0.000 description 4
- 238000012545 processing Methods 0.000 description 4
- 230000010076 replication Effects 0.000 description 4
- 238000012360 testing method Methods 0.000 description 4
- 238000011282 treatment Methods 0.000 description 4
- 102100021933 C-C motif chemokine 25 Human genes 0.000 description 3
- 102100021936 C-C motif chemokine 27 Human genes 0.000 description 3
- 101710112538 C-C motif chemokine 27 Proteins 0.000 description 3
- 102100021942 C-C motif chemokine 28 Human genes 0.000 description 3
- 101000897486 Homo sapiens C-C motif chemokine 25 Proteins 0.000 description 3
- 101000897477 Homo sapiens C-C motif chemokine 28 Proteins 0.000 description 3
- 101000679903 Homo sapiens Tumor necrosis factor receptor superfamily member 25 Proteins 0.000 description 3
- 102000013462 Interleukin-12 Human genes 0.000 description 3
- 108010065805 Interleukin-12 Proteins 0.000 description 3
- 102000003812 Interleukin-15 Human genes 0.000 description 3
- 108090000172 Interleukin-15 Proteins 0.000 description 3
- 108090000978 Interleukin-4 Proteins 0.000 description 3
- 102000004388 Interleukin-4 Human genes 0.000 description 3
- 241000714474 Rous sarcoma virus Species 0.000 description 3
- 210000001744 T-lymphocyte Anatomy 0.000 description 3
- 102100040112 Tumor necrosis factor receptor superfamily member 10B Human genes 0.000 description 3
- 238000013459 approach Methods 0.000 description 3
- 230000008901 benefit Effects 0.000 description 3
- 210000000349 chromosome Anatomy 0.000 description 3
- 230000001186 cumulative effect Effects 0.000 description 3
- 239000003814 drug Substances 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- 230000004927 fusion Effects 0.000 description 3
- 206010073071 hepatocellular carcinoma Diseases 0.000 description 3
- 231100000844 hepatocellular carcinoma Toxicity 0.000 description 3
- 238000010166 immunofluorescence Methods 0.000 description 3
- 238000001727 in vivo Methods 0.000 description 3
- 230000035772 mutation Effects 0.000 description 3
- 230000001681 protective effect Effects 0.000 description 3
- 230000006337 proteolytic cleavage Effects 0.000 description 3
- 230000000241 respiratory effect Effects 0.000 description 3
- 230000035945 sensitivity Effects 0.000 description 3
- 239000007858 starting material Substances 0.000 description 3
- 230000003612 virological effect Effects 0.000 description 3
- DIGQNXIGRZPYDK-WKSCXVIASA-N (2R)-6-amino-2-[[2-[[(2S)-2-[[2-[[(2R)-2-[[(2S)-2-[[(2R,3S)-2-[[2-[[(2S)-2-[[2-[[(2S)-2-[[(2S)-2-[[(2R)-2-[[(2S,3S)-2-[[(2R)-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[2-[[(2S)-2-[[(2R)-2-[[2-[[2-[[2-[(2-amino-1-hydroxyethylidene)amino]-3-carboxy-1-hydroxypropylidene]amino]-1-hydroxy-3-sulfanylpropylidene]amino]-1-hydroxyethylidene]amino]-1-hydroxy-3-sulfanylpropylidene]amino]-1,3-dihydroxypropylidene]amino]-1-hydroxyethylidene]amino]-1-hydroxypropylidene]amino]-1,3-dihydroxypropylidene]amino]-1,3-dihydroxypropylidene]amino]-1-hydroxy-3-sulfanylpropylidene]amino]-1,3-dihydroxybutylidene]amino]-1-hydroxy-3-sulfanylpropylidene]amino]-1-hydroxypropylidene]amino]-1,3-dihydroxypropylidene]amino]-1-hydroxyethylidene]amino]-1,5-dihydroxy-5-iminopentylidene]amino]-1-hydroxy-3-sulfanylpropylidene]amino]-1,3-dihydroxybutylidene]amino]-1-hydroxy-3-sulfanylpropylidene]amino]-1,3-dihydroxypropylidene]amino]-1-hydroxyethylidene]amino]-1-hydroxy-3-sulfanylpropylidene]amino]-1-hydroxyethylidene]amino]hexanoic acid Chemical compound C[C@@H]([C@@H](C(=N[C@@H](CS)C(=N[C@@H](C)C(=N[C@@H](CO)C(=NCC(=N[C@@H](CCC(=N)O)C(=NC(CS)C(=N[C@H]([C@H](C)O)C(=N[C@H](CS)C(=N[C@H](CO)C(=NCC(=N[C@H](CS)C(=NCC(=N[C@H](CCCCN)C(=O)O)O)O)O)O)O)O)O)O)O)O)O)O)O)N=C([C@H](CS)N=C([C@H](CO)N=C([C@H](CO)N=C([C@H](C)N=C(CN=C([C@H](CO)N=C([C@H](CS)N=C(CN=C(C(CS)N=C(C(CC(=O)O)N=C(CN)O)O)O)O)O)O)O)O)O)O)O)O DIGQNXIGRZPYDK-WKSCXVIASA-N 0.000 description 2
- YYGNTYWPHWGJRM-UHFFFAOYSA-N (6E,10E,14E,18E)-2,6,10,15,19,23-hexamethyltetracosa-2,6,10,14,18,22-hexaene Chemical compound CC(C)=CCCC(C)=CCCC(C)=CCCC=C(C)CCC=C(C)CCC=C(C)C YYGNTYWPHWGJRM-UHFFFAOYSA-N 0.000 description 2
- 102000007469 Actins Human genes 0.000 description 2
- 108010085238 Actins Proteins 0.000 description 2
- 241000714230 Avian leukemia virus Species 0.000 description 2
- 241000713704 Bovine immunodeficiency virus Species 0.000 description 2
- 102000004127 Cytokines Human genes 0.000 description 2
- 108090000695 Cytokines Proteins 0.000 description 2
- 241000588724 Escherichia coli Species 0.000 description 2
- ZHNUHDYFZUAESO-UHFFFAOYSA-N Formamide Chemical compound NC=O ZHNUHDYFZUAESO-UHFFFAOYSA-N 0.000 description 2
- 102000003886 Glycoproteins Human genes 0.000 description 2
- 108090000288 Glycoproteins Proteins 0.000 description 2
- 108010017213 Granulocyte-Macrophage Colony-Stimulating Factor Proteins 0.000 description 2
- 102100039620 Granulocyte-macrophage colony-stimulating factor Human genes 0.000 description 2
- 241000560067 HIV-1 group M Species 0.000 description 2
- 102000001554 Hemoglobins Human genes 0.000 description 2
- 108010054147 Hemoglobins Proteins 0.000 description 2
- 241000282412 Homo Species 0.000 description 2
- 101000610604 Homo sapiens Tumor necrosis factor receptor superfamily member 10B Proteins 0.000 description 2
- 241000725303 Human immunodeficiency virus Species 0.000 description 2
- 108060003951 Immunoglobulin Proteins 0.000 description 2
- 108010047761 Interferon-alpha Proteins 0.000 description 2
- 102000006992 Interferon-alpha Human genes 0.000 description 2
- 102000008070 Interferon-gamma Human genes 0.000 description 2
- 108010002352 Interleukin-1 Proteins 0.000 description 2
- 102000000589 Interleukin-1 Human genes 0.000 description 2
- 102000003814 Interleukin-10 Human genes 0.000 description 2
- 108090000174 Interleukin-10 Proteins 0.000 description 2
- 102000003810 Interleukin-18 Human genes 0.000 description 2
- 108090000171 Interleukin-18 Proteins 0.000 description 2
- 108010002350 Interleukin-2 Proteins 0.000 description 2
- 102000000588 Interleukin-2 Human genes 0.000 description 2
- 108090001005 Interleukin-6 Proteins 0.000 description 2
- 102000004889 Interleukin-6 Human genes 0.000 description 2
- 108010074338 Lymphokines Proteins 0.000 description 2
- 102000008072 Lymphokines Human genes 0.000 description 2
- 102000003792 Metallothionein Human genes 0.000 description 2
- 108090000157 Metallothionein Proteins 0.000 description 2
- 102000003505 Myosin Human genes 0.000 description 2
- 108060008487 Myosin Proteins 0.000 description 2
- 101710111275 Non-structural 3 Proteins 0.000 description 2
- 108091034117 Oligonucleotide Proteins 0.000 description 2
- 208000037581 Persistent Infection Diseases 0.000 description 2
- 206010035148 Plague Diseases 0.000 description 2
- 108010008281 Recombinant Fusion Proteins Proteins 0.000 description 2
- 102000007056 Recombinant Fusion Proteins Human genes 0.000 description 2
- IWUCXVSUMQZMFG-AFCXAGJDSA-N Ribavirin Chemical compound N1=C(C(=O)N)N=CN1[C@H]1[C@H](O)[C@H](O)[C@@H](CO)O1 IWUCXVSUMQZMFG-AFCXAGJDSA-N 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 108091081024 Start codon Proteins 0.000 description 2
- 101150056647 TNFRSF4 gene Proteins 0.000 description 2
- BHEOSNUKNHRBNM-UHFFFAOYSA-N Tetramethylsqualene Natural products CC(=C)C(C)CCC(=C)C(C)CCC(C)=CCCC=C(C)CCC(C)C(=C)CCC(C)C(C)=C BHEOSNUKNHRBNM-UHFFFAOYSA-N 0.000 description 2
- 241000607479 Yersinia pestis Species 0.000 description 2
- JLCPHMBAVCMARE-UHFFFAOYSA-N [3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-[[3-[[3-[[3-[[3-[[3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-hydroxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methyl [5-(6-aminopurin-9-yl)-2-(hydroxymethyl)oxolan-3-yl] hydrogen phosphate Polymers Cc1cn(C2CC(OP(O)(=O)OCC3OC(CC3OP(O)(=O)OCC3OC(CC3O)n3cnc4c3nc(N)[nH]c4=O)n3cnc4c3nc(N)[nH]c4=O)C(COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3CO)n3cnc4c(N)ncnc34)n3ccc(N)nc3=O)n3cnc4c(N)ncnc34)n3ccc(N)nc3=O)n3ccc(N)nc3=O)n3ccc(N)nc3=O)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cc(C)c(=O)[nH]c3=O)n3cc(C)c(=O)[nH]c3=O)n3ccc(N)nc3=O)n3cc(C)c(=O)[nH]c3=O)n3cnc4c3nc(N)[nH]c4=O)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)O2)c(=O)[nH]c1=O JLCPHMBAVCMARE-UHFFFAOYSA-N 0.000 description 2
- 231100000354 acute hepatitis Toxicity 0.000 description 2
- 230000003044 adaptive effect Effects 0.000 description 2
- 239000002671 adjuvant Substances 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 210000004369 blood Anatomy 0.000 description 2
- 239000008280 blood Substances 0.000 description 2
- 229940030156 cell vaccine Drugs 0.000 description 2
- 230000001413 cellular effect Effects 0.000 description 2
- 238000004891 communication Methods 0.000 description 2
- 229960003624 creatine Drugs 0.000 description 2
- 239000006046 creatine Substances 0.000 description 2
- 210000001151 cytotoxic T lymphocyte Anatomy 0.000 description 2
- 230000006378 damage Effects 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- PRAKJMSDJKAYCZ-UHFFFAOYSA-N dodecahydrosqualene Natural products CC(C)CCCC(C)CCCC(C)CCCCC(C)CCCC(C)CCCC(C)C PRAKJMSDJKAYCZ-UHFFFAOYSA-N 0.000 description 2
- 229940079593 drug Drugs 0.000 description 2
- 238000009472 formulation Methods 0.000 description 2
- 229940044627 gamma-interferon Drugs 0.000 description 2
- 238000010353 genetic engineering Methods 0.000 description 2
- 239000003102 growth factor Substances 0.000 description 2
- 230000005802 health problem Effects 0.000 description 2
- 231100000283 hepatitis Toxicity 0.000 description 2
- 210000003494 hepatocyte Anatomy 0.000 description 2
- 210000000987 immune system Anatomy 0.000 description 2
- 102000018358 immunoglobulin Human genes 0.000 description 2
- 230000001024 immunotherapeutic effect Effects 0.000 description 2
- 230000002458 infectious effect Effects 0.000 description 2
- 230000000977 initiatory effect Effects 0.000 description 2
- 238000002955 isolation Methods 0.000 description 2
- 208000019423 liver disease Diseases 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 239000011159 matrix material Substances 0.000 description 2
- 239000002609 medium Substances 0.000 description 2
- 239000013642 negative control Substances 0.000 description 2
- 230000002688 persistence Effects 0.000 description 2
- 108091033319 polynucleotide Proteins 0.000 description 2
- 102000040430 polynucleotide Human genes 0.000 description 2
- 239000002157 polynucleotide Substances 0.000 description 2
- 230000000750 progressive effect Effects 0.000 description 2
- 108020001775 protein parts Proteins 0.000 description 2
- 238000011084 recovery Methods 0.000 description 2
- 229960000329 ribavirin Drugs 0.000 description 2
- HZCAHMRRMINHDJ-DBRKOABJSA-N ribavirin Natural products O[C@@H]1[C@H](O)[C@@H](CO)O[C@H]1N1N=CN=C1 HZCAHMRRMINHDJ-DBRKOABJSA-N 0.000 description 2
- 150000003839 salts Chemical class 0.000 description 2
- 229910001415 sodium ion Inorganic materials 0.000 description 2
- 210000000952 spleen Anatomy 0.000 description 2
- 229940031439 squalene Drugs 0.000 description 2
- TUHBEKDERLKLEC-UHFFFAOYSA-N squalene Natural products CC(=CCCC(=CCCC(=CCCC=C(/C)CCC=C(/C)CC=C(C)C)C)C)C TUHBEKDERLKLEC-UHFFFAOYSA-N 0.000 description 2
- 238000011272 standard treatment Methods 0.000 description 2
- 238000007920 subcutaneous administration Methods 0.000 description 2
- 238000001890 transfection Methods 0.000 description 2
- 238000002054 transplantation Methods 0.000 description 2
- 241000701161 unidentified adenovirus Species 0.000 description 2
- VBEQCZHXXJYVRD-GACYYNSASA-N uroanthelone Chemical compound C([C@@H](C(=O)N[C@H](C(=O)N[C@@H](CS)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CS)C(=O)N[C@H](C(=O)N[C@@H]([C@@H](C)CC)C(=O)NCC(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)N[C@@H](CO)C(=O)NCC(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CS)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O)C(C)C)[C@@H](C)O)NC(=O)[C@H](CO)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CO)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@@H](NC(=O)[C@H](CC=1NC=NC=1)NC(=O)[C@H](CCSC)NC(=O)[C@H](CS)NC(=O)[C@@H](NC(=O)CNC(=O)CNC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CS)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)CNC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CO)NC(=O)[C@H](CO)NC(=O)[C@H]1N(CCC1)C(=O)[C@H](CS)NC(=O)CNC(=O)[C@H]1N(CCC1)C(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CO)NC(=O)[C@@H](N)CC(N)=O)C(C)C)[C@@H](C)CC)C1=CC=C(O)C=C1 VBEQCZHXXJYVRD-GACYYNSASA-N 0.000 description 2
- 238000002255 vaccination Methods 0.000 description 2
- 230000009385 viral infection Effects 0.000 description 2
- 229960004854 viral vaccine Drugs 0.000 description 2
- KIUKXJAPPMFGSW-DNGZLQJQSA-N (2S,3S,4S,5R,6R)-6-[(2S,3R,4R,5S,6R)-3-Acetamido-2-[(2S,3S,4R,5R,6R)-6-[(2R,3R,4R,5S,6R)-3-acetamido-2,5-dihydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-2-carboxy-4,5-dihydroxyoxan-3-yl]oxy-5-hydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-3,4,5-trihydroxyoxane-2-carboxylic acid Chemical compound CC(=O)N[C@H]1[C@H](O)O[C@H](CO)[C@@H](O)[C@@H]1O[C@H]1[C@H](O)[C@@H](O)[C@H](O[C@H]2[C@@H]([C@@H](O[C@H]3[C@@H]([C@@H](O)[C@H](O)[C@H](O3)C(O)=O)O)[C@H](O)[C@@H](CO)O2)NC(C)=O)[C@@H](C(O)=O)O1 KIUKXJAPPMFGSW-DNGZLQJQSA-N 0.000 description 1
- 206010065051 Acute hepatitis C Diseases 0.000 description 1
- 102000009027 Albumins Human genes 0.000 description 1
- 108010088751 Albumins Proteins 0.000 description 1
- GUBGYTABKSRVRQ-XLOQQCSPSA-N Alpha-Lactose Chemical compound O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@@H](CO)O[C@H](O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-XLOQQCSPSA-N 0.000 description 1
- 240000001546 Byrsonima crassifolia Species 0.000 description 1
- 235000003197 Byrsonima crassifolia Nutrition 0.000 description 1
- 101710155857 C-C motif chemokine 2 Proteins 0.000 description 1
- 102100021943 C-C motif chemokine 2 Human genes 0.000 description 1
- 102100032367 C-C motif chemokine 5 Human genes 0.000 description 1
- 108700012434 CCL3 Proteins 0.000 description 1
- 108010029697 CD40 Ligand Proteins 0.000 description 1
- 101150013553 CD40 gene Proteins 0.000 description 1
- 102100032937 CD40 ligand Human genes 0.000 description 1
- 108010084313 CD58 Antigens Proteins 0.000 description 1
- 101100289995 Caenorhabditis elegans mac-1 gene Proteins 0.000 description 1
- 241000283707 Capra Species 0.000 description 1
- 102000011727 Caspases Human genes 0.000 description 1
- 108010076667 Caspases Proteins 0.000 description 1
- 102000000013 Chemokine CCL3 Human genes 0.000 description 1
- 108010055166 Chemokine CCL5 Proteins 0.000 description 1
- 206010008909 Chronic Hepatitis Diseases 0.000 description 1
- KRKNYBCHXYNGOX-UHFFFAOYSA-K Citrate Chemical compound [O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O KRKNYBCHXYNGOX-UHFFFAOYSA-K 0.000 description 1
- 229920000832 Cutin Polymers 0.000 description 1
- FBPFZTCFMRRESA-FSIIMWSLSA-N D-Glucitol Natural products OC[C@H](O)[C@H](O)[C@@H](O)[C@H](O)CO FBPFZTCFMRRESA-FSIIMWSLSA-N 0.000 description 1
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 description 1
- FBPFZTCFMRRESA-JGWLITMVSA-N D-glucitol Chemical compound OC[C@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-JGWLITMVSA-N 0.000 description 1
- 238000011238 DNA vaccination Methods 0.000 description 1
- 101710088341 Dermatopontin Proteins 0.000 description 1
- 101100347633 Drosophila melanogaster Mhc gene Proteins 0.000 description 1
- 108010024212 E-Selectin Proteins 0.000 description 1
- 102100023471 E-selectin Human genes 0.000 description 1
- 108010031111 EBV-encoded nuclear antigen 1 Proteins 0.000 description 1
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 1
- 101710130332 ETS domain-containing protein Elk-4 Proteins 0.000 description 1
- 102000018233 Fibroblast Growth Factor Human genes 0.000 description 1
- 108050007372 Fibroblast Growth Factor Proteins 0.000 description 1
- 206010016654 Fibrosis Diseases 0.000 description 1
- 229920001917 Ficoll Polymers 0.000 description 1
- 101710177291 Gag polyprotein Proteins 0.000 description 1
- 108010010803 Gelatin Proteins 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- 102100031573 Hematopoietic progenitor cell antigen CD34 Human genes 0.000 description 1
- 206010019799 Hepatitis viral Diseases 0.000 description 1
- 241000238631 Hexapoda Species 0.000 description 1
- 101000777663 Homo sapiens Hematopoietic progenitor cell antigen CD34 Proteins 0.000 description 1
- 101000991061 Homo sapiens MHC class I polypeptide-related sequence B Proteins 0.000 description 1
- 101001109501 Homo sapiens NKG2-D type II integral membrane protein Proteins 0.000 description 1
- 101000934346 Homo sapiens T-cell surface antigen CD2 Proteins 0.000 description 1
- 101000914484 Homo sapiens T-lymphocyte activation antigen CD80 Proteins 0.000 description 1
- 101100100117 Homo sapiens TNFRSF10B gene Proteins 0.000 description 1
- 101001050288 Homo sapiens Transcription factor Jun Proteins 0.000 description 1
- 101000611183 Homo sapiens Tumor necrosis factor Proteins 0.000 description 1
- 101000610602 Homo sapiens Tumor necrosis factor receptor superfamily member 10C Proteins 0.000 description 1
- 101000610609 Homo sapiens Tumor necrosis factor receptor superfamily member 10D Proteins 0.000 description 1
- XQFRJNBWHJMXHO-RRKCRQDMSA-N IDUR Chemical compound C1[C@H](O)[C@@H](CO)O[C@H]1N1C(=O)NC(=O)C(I)=C1 XQFRJNBWHJMXHO-RRKCRQDMSA-N 0.000 description 1
- 208000002979 Influenza in Birds Diseases 0.000 description 1
- 102100025323 Integrin alpha-1 Human genes 0.000 description 1
- 102100022339 Integrin alpha-L Human genes 0.000 description 1
- 108010041341 Integrin alpha1 Proteins 0.000 description 1
- 108010055795 Integrin alpha1beta1 Proteins 0.000 description 1
- 108010064593 Intercellular Adhesion Molecule-1 Proteins 0.000 description 1
- 108010064600 Intercellular Adhesion Molecule-3 Proteins 0.000 description 1
- 102100037877 Intercellular adhesion molecule 1 Human genes 0.000 description 1
- 101710148794 Intercellular adhesion molecule 2 Proteins 0.000 description 1
- 102100037872 Intercellular adhesion molecule 2 Human genes 0.000 description 1
- 102100037871 Intercellular adhesion molecule 3 Human genes 0.000 description 1
- 108010078049 Interferon alpha-2 Proteins 0.000 description 1
- 102100036342 Interleukin-1 receptor-associated kinase 1 Human genes 0.000 description 1
- 101710199015 Interleukin-1 receptor-associated kinase 1 Proteins 0.000 description 1
- 102100026878 Interleukin-2 receptor subunit alpha Human genes 0.000 description 1
- 108010002616 Interleukin-5 Proteins 0.000 description 1
- 102000000743 Interleukin-5 Human genes 0.000 description 1
- 108010002586 Interleukin-7 Proteins 0.000 description 1
- 102000000704 Interleukin-7 Human genes 0.000 description 1
- 108090001007 Interleukin-8 Proteins 0.000 description 1
- 102000004890 Interleukin-8 Human genes 0.000 description 1
- 108020003285 Isocitrate lyase Proteins 0.000 description 1
- 108010055717 JNK Mitogen-Activated Protein Kinases Proteins 0.000 description 1
- 102000019145 JUN kinase activity proteins Human genes 0.000 description 1
- 101150069255 KLRC1 gene Proteins 0.000 description 1
- YQEZLKZALYSWHR-UHFFFAOYSA-N Ketamine Chemical compound C=1C=CC=C(Cl)C=1C1(NC)CCCCC1=O YQEZLKZALYSWHR-UHFFFAOYSA-N 0.000 description 1
- 108010092694 L-Selectin Proteins 0.000 description 1
- ZDXPYRJPNDTMRX-VKHMYHEASA-N L-glutamine Chemical compound OC(=O)[C@@H](N)CCC(N)=O ZDXPYRJPNDTMRX-VKHMYHEASA-N 0.000 description 1
- 229930182816 L-glutamine Natural products 0.000 description 1
- 102100033467 L-selectin Human genes 0.000 description 1
- GUBGYTABKSRVRQ-QKKXKWKRSA-N Lactose Natural products OC[C@H]1O[C@@H](O[C@H]2[C@H](O)[C@@H](O)C(O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@H]1O GUBGYTABKSRVRQ-QKKXKWKRSA-N 0.000 description 1
- 108010064548 Lymphocyte Function-Associated Antigen-1 Proteins 0.000 description 1
- 102100030301 MHC class I polypeptide-related sequence A Human genes 0.000 description 1
- 102100030300 MHC class I polypeptide-related sequence B Human genes 0.000 description 1
- 241000282567 Macaca fascicularis Species 0.000 description 1
- 101100404845 Macaca mulatta NKG2A gene Proteins 0.000 description 1
- 108010046938 Macrophage Colony-Stimulating Factor Proteins 0.000 description 1
- 102100028123 Macrophage colony-stimulating factor 1 Human genes 0.000 description 1
- 101710125418 Major capsid protein Proteins 0.000 description 1
- 241000124008 Mammalia Species 0.000 description 1
- 229930195725 Mannitol Natural products 0.000 description 1
- 108010060408 Member 25 Tumor Necrosis Factor Receptors Proteins 0.000 description 1
- 241000713333 Mouse mammary tumor virus Species 0.000 description 1
- 101100372761 Mus musculus Flt1 gene Proteins 0.000 description 1
- 101001044384 Mus musculus Interferon gamma Proteins 0.000 description 1
- 102000010168 Myeloid Differentiation Factor 88 Human genes 0.000 description 1
- 108010077432 Myeloid Differentiation Factor 88 Proteins 0.000 description 1
- 102100022682 NKG2-A/NKG2-B type II integral membrane protein Human genes 0.000 description 1
- 102100022680 NKG2-D type II integral membrane protein Human genes 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 108010025020 Nerve Growth Factor Proteins 0.000 description 1
- 102000015336 Nerve Growth Factor Human genes 0.000 description 1
- 108700001237 Nucleic Acid-Based Vaccines Proteins 0.000 description 1
- 108700020796 Oncogene Proteins 0.000 description 1
- 108700026244 Open Reading Frames Proteins 0.000 description 1
- 108010035766 P-Selectin Proteins 0.000 description 1
- 102100023472 P-selectin Human genes 0.000 description 1
- 101150044441 PECAM1 gene Proteins 0.000 description 1
- 241000288906 Primates Species 0.000 description 1
- 102100028688 Putative glycosylation-dependent cell adhesion molecule 1 Human genes 0.000 description 1
- 108010025832 RANK Ligand Proteins 0.000 description 1
- 102000014128 RANK Ligand Human genes 0.000 description 1
- 239000012979 RPMI medium Substances 0.000 description 1
- 239000012980 RPMI-1640 medium Substances 0.000 description 1
- 101710138742 Receptor-type tyrosine-protein phosphatase H Proteins 0.000 description 1
- 108020004511 Recombinant DNA Proteins 0.000 description 1
- 208000035415 Reinfection Diseases 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- 102400000830 Saposin-B Human genes 0.000 description 1
- 241000282887 Suidae Species 0.000 description 1
- 102100025237 T-cell surface antigen CD2 Human genes 0.000 description 1
- 102100027222 T-lymphocyte activation antigen CD80 Human genes 0.000 description 1
- 102000003714 TNF receptor-associated factor 6 Human genes 0.000 description 1
- 108090000009 TNF receptor-associated factor 6 Proteins 0.000 description 1
- 239000004098 Tetracycline Substances 0.000 description 1
- 102000006601 Thymidine Kinase Human genes 0.000 description 1
- 108020004440 Thymidine kinase Proteins 0.000 description 1
- 102100023132 Transcription factor Jun Human genes 0.000 description 1
- 108060008682 Tumor Necrosis Factor Proteins 0.000 description 1
- 108060008683 Tumor Necrosis Factor Receptor Proteins 0.000 description 1
- 102000000852 Tumor Necrosis Factor-alpha Human genes 0.000 description 1
- 102100040247 Tumor necrosis factor Human genes 0.000 description 1
- 102100040115 Tumor necrosis factor receptor superfamily member 10C Human genes 0.000 description 1
- 102100040110 Tumor necrosis factor receptor superfamily member 10D Human genes 0.000 description 1
- 102100040245 Tumor necrosis factor receptor superfamily member 5 Human genes 0.000 description 1
- 108010073929 Vascular Endothelial Growth Factor A Proteins 0.000 description 1
- 102000005789 Vascular Endothelial Growth Factors Human genes 0.000 description 1
- 108010019530 Vascular Endothelial Growth Factors Proteins 0.000 description 1
- 208000036142 Viral infection Diseases 0.000 description 1
- NOSIYYJFMPDDSA-UHFFFAOYSA-N acepromazine Chemical compound C1=C(C(C)=O)C=C2N(CCCN(C)C)C3=CC=CC=C3SC2=C1 NOSIYYJFMPDDSA-UHFFFAOYSA-N 0.000 description 1
- 229960005054 acepromazine Drugs 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 150000007513 acids Chemical class 0.000 description 1
- 230000004721 adaptive immunity Effects 0.000 description 1
- 239000000654 additive Substances 0.000 description 1
- 238000013019 agitation Methods 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
- 150000001450 anions Chemical class 0.000 description 1
- 230000001857 anti-mycotic effect Effects 0.000 description 1
- 239000002543 antimycotic Substances 0.000 description 1
- 238000003491 array Methods 0.000 description 1
- MXWJVTOOROXGIU-UHFFFAOYSA-N atrazine Chemical compound CCNC1=NC(Cl)=NC(NC(C)C)=N1 MXWJVTOOROXGIU-UHFFFAOYSA-N 0.000 description 1
- 206010064097 avian influenza Diseases 0.000 description 1
- 238000002869 basic local alignment search tool Methods 0.000 description 1
- 230000006399 behavior Effects 0.000 description 1
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 1
- 230000003115 biocidal effect Effects 0.000 description 1
- 230000005540 biological transmission Effects 0.000 description 1
- 231100000504 carcinogenesis Toxicity 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 230000007969 cellular immunity Effects 0.000 description 1
- 210000002230 centromere Anatomy 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 210000004978 chinese hamster ovary cell Anatomy 0.000 description 1
- 239000013611 chromosomal DNA Substances 0.000 description 1
- 208000020403 chronic hepatitis C virus infection Diseases 0.000 description 1
- 230000007882 cirrhosis Effects 0.000 description 1
- 208000019425 cirrhosis of liver Diseases 0.000 description 1
- 238000003776 cleavage reaction Methods 0.000 description 1
- 238000010367 cloning Methods 0.000 description 1
- 230000000295 complement effect Effects 0.000 description 1
- 238000012790 confirmation Methods 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 230000009089 cytolysis Effects 0.000 description 1
- 230000003013 cytotoxicity Effects 0.000 description 1
- 231100000135 cytotoxicity Toxicity 0.000 description 1
- 230000034994 death Effects 0.000 description 1
- 238000000432 density-gradient centrifugation Methods 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 230000000368 destabilizing effect Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 238000002405 diagnostic procedure Methods 0.000 description 1
- LOKCTEFSRHRXRJ-UHFFFAOYSA-I dipotassium trisodium dihydrogen phosphate hydrogen phosphate dichloride Chemical compound P(=O)(O)(O)[O-].[K+].P(=O)(O)([O-])[O-].[Na+].[Na+].[Cl-].[K+].[Cl-].[Na+] LOKCTEFSRHRXRJ-UHFFFAOYSA-I 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 241001493065 dsRNA viruses Species 0.000 description 1
- 238000001962 electrophoresis Methods 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 230000010502 episomal replication Effects 0.000 description 1
- 210000003743 erythrocyte Anatomy 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 239000013613 expression plasmid Substances 0.000 description 1
- 229940126864 fibroblast growth factor Drugs 0.000 description 1
- 229960002963 ganciclovir Drugs 0.000 description 1
- IRSCQMHQWWYFCW-UHFFFAOYSA-N ganciclovir Chemical compound O=C1NC(N)=NC2=C1N=CN2COC(CO)CO IRSCQMHQWWYFCW-UHFFFAOYSA-N 0.000 description 1
- 229920000159 gelatin Polymers 0.000 description 1
- 239000008273 gelatin Substances 0.000 description 1
- 235000019322 gelatine Nutrition 0.000 description 1
- 235000011852 gelatine desserts Nutrition 0.000 description 1
- 238000012239 gene modification Methods 0.000 description 1
- 230000005017 genetic modification Effects 0.000 description 1
- 235000013617 genetically modified food Nutrition 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 210000002443 helper t lymphocyte Anatomy 0.000 description 1
- 208000010710 hepatitis C virus infection Diseases 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- 229920002674 hyaluronan Polymers 0.000 description 1
- 229960003160 hyaluronic acid Drugs 0.000 description 1
- 238000009396 hybridization Methods 0.000 description 1
- 230000002519 immonomodulatory effect Effects 0.000 description 1
- 230000004957 immunoregulator effect Effects 0.000 description 1
- 238000009169 immunotherapy Methods 0.000 description 1
- 230000006698 induction Effects 0.000 description 1
- 230000002757 inflammatory effect Effects 0.000 description 1
- 238000011221 initial treatment Methods 0.000 description 1
- 229960003507 interferon alfa-2b Drugs 0.000 description 1
- 230000010468 interferon response Effects 0.000 description 1
- 238000001361 intraarterial administration Methods 0.000 description 1
- 238000007918 intramuscular administration Methods 0.000 description 1
- 238000007912 intraperitoneal administration Methods 0.000 description 1
- 239000007928 intraperitoneal injection Substances 0.000 description 1
- 238000001990 intravenous administration Methods 0.000 description 1
- 239000000644 isotonic solution Substances 0.000 description 1
- 229960003299 ketamine Drugs 0.000 description 1
- 239000008101 lactose Substances 0.000 description 1
- 208000032839 leukemia Diseases 0.000 description 1
- 239000003446 ligand Substances 0.000 description 1
- 239000002502 liposome Substances 0.000 description 1
- 201000007270 liver cancer Diseases 0.000 description 1
- 208000014018 liver neoplasm Diseases 0.000 description 1
- 210000004698 lymphocyte Anatomy 0.000 description 1
- 230000002934 lysing effect Effects 0.000 description 1
- 239000012139 lysis buffer Substances 0.000 description 1
- 210000004962 mammalian cell Anatomy 0.000 description 1
- 238000007726 management method Methods 0.000 description 1
- 239000000594 mannitol Substances 0.000 description 1
- 235000010355 mannitol Nutrition 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 238000002844 melting Methods 0.000 description 1
- 230000008018 melting Effects 0.000 description 1
- 210000003071 memory t lymphocyte Anatomy 0.000 description 1
- 108020004999 messenger RNA Proteins 0.000 description 1
- 239000010445 mica Substances 0.000 description 1
- 229910052618 mica group Inorganic materials 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 239000011859 microparticle Substances 0.000 description 1
- 238000010369 molecular cloning Methods 0.000 description 1
- 229940035032 monophosphoryl lipid a Drugs 0.000 description 1
- 229940126619 mouse monoclonal antibody Drugs 0.000 description 1
- 230000016379 mucosal immune response Effects 0.000 description 1
- 210000004400 mucous membrane Anatomy 0.000 description 1
- 125000001446 muramyl group Chemical group N[C@@H](C=O)[C@@H](O[C@@H](C(=O)*)C)[C@H](O)[C@H](O)CO 0.000 description 1
- 229940053128 nerve growth factor Drugs 0.000 description 1
- 230000003472 neutralizing effect Effects 0.000 description 1
- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- 229940023146 nucleic acid vaccine Drugs 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 230000001717 pathogenic effect Effects 0.000 description 1
- 230000007918 pathogenicity Effects 0.000 description 1
- 229940021222 peritoneal dialysis isotonic solution Drugs 0.000 description 1
- 230000002085 persistent effect Effects 0.000 description 1
- 239000002953 phosphate buffered saline Substances 0.000 description 1
- 238000013081 phylogenetic analysis Methods 0.000 description 1
- 229920002643 polyglutamic acid Polymers 0.000 description 1
- 238000004393 prognosis Methods 0.000 description 1
- 230000002250 progressing effect Effects 0.000 description 1
- 230000000069 prophylactic effect Effects 0.000 description 1
- 229940021993 prophylactic vaccine Drugs 0.000 description 1
- 108020003519 protein disulfide isomerase Proteins 0.000 description 1
- 229940023143 protein vaccine Drugs 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 239000002510 pyrogen Substances 0.000 description 1
- 238000004445 quantitative analysis Methods 0.000 description 1
- 150000004053 quinones Chemical class 0.000 description 1
- 230000035484 reaction time Effects 0.000 description 1
- 238000010188 recombinant method Methods 0.000 description 1
- 210000000664 rectum Anatomy 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 108091008146 restriction endonucleases Proteins 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- MIXCUJKCXRNYFM-UHFFFAOYSA-M sodium;diiodomethanesulfonate;n-propyl-n-[2-(2,4,6-trichlorophenoxy)ethyl]imidazole-1-carboxamide Chemical compound [Na+].[O-]S(=O)(=O)C(I)I.C1=CN=CN1C(=O)N(CCC)CCOC1=C(Cl)C=C(Cl)C=C1Cl MIXCUJKCXRNYFM-UHFFFAOYSA-M 0.000 description 1
- 239000000600 sorbitol Substances 0.000 description 1
- 239000003381 stabilizer Substances 0.000 description 1
- 229910001220 stainless steel Inorganic materials 0.000 description 1
- 239000010935 stainless steel Substances 0.000 description 1
- 238000007619 statistical method Methods 0.000 description 1
- 230000004936 stimulating effect Effects 0.000 description 1
- 238000010254 subcutaneous injection Methods 0.000 description 1
- 239000007929 subcutaneous injection Substances 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 108010012704 sulfated glycoprotein p50 Proteins 0.000 description 1
- 239000000829 suppository Substances 0.000 description 1
- 239000004094 surface-active agent Substances 0.000 description 1
- 210000003411 telomere Anatomy 0.000 description 1
- 102000055501 telomere Human genes 0.000 description 1
- 108091035539 telomere Proteins 0.000 description 1
- 229960002180 tetracycline Drugs 0.000 description 1
- 229930101283 tetracycline Natural products 0.000 description 1
- 235000019364 tetracycline Nutrition 0.000 description 1
- 150000003522 tetracyclines Chemical class 0.000 description 1
- JGVWCANSWKRBCS-UHFFFAOYSA-N tetramethylrhodamine thiocyanate Chemical compound [Cl-].C=12C=CC(N(C)C)=CC2=[O+]C2=CC(N(C)C)=CC=C2C=1C1=CC=C(SC#N)C=C1C(O)=O JGVWCANSWKRBCS-UHFFFAOYSA-N 0.000 description 1
- 229940021747 therapeutic vaccine Drugs 0.000 description 1
- 238000004448 titration Methods 0.000 description 1
- 230000000699 topical effect Effects 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 238000003146 transient transfection Methods 0.000 description 1
- 102000003298 tumor necrosis factor receptor Human genes 0.000 description 1
- 241000701447 unidentified baculovirus Species 0.000 description 1
- 238000011144 upstream manufacturing Methods 0.000 description 1
- 210000003708 urethra Anatomy 0.000 description 1
- 210000001215 vagina Anatomy 0.000 description 1
- 230000006444 vascular growth Effects 0.000 description 1
- 239000005526 vasoconstrictor agent Substances 0.000 description 1
- 230000029812 viral genome replication Effects 0.000 description 1
- 201000001862 viral hepatitis Diseases 0.000 description 1
- 230000006514 viral protein processing Effects 0.000 description 1
- 239000013603 viral vector Substances 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
- DGVVWUTYPXICAM-UHFFFAOYSA-N β‐Mercaptoethanol Chemical compound OCCS DGVVWUTYPXICAM-UHFFFAOYSA-N 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H21/00—Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/7088—Compounds having three or more nucleosides or nucleotides
- A61K31/711—Natural deoxyribonucleic acids, i.e. containing only 2'-deoxyriboses attached to adenine, guanine, cytosine or thymine and having 3'-5' phosphodiester links
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/12—Viral antigens
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/12—Viral antigens
- A61K39/29—Hepatitis virus
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/12—Antivirals
- A61P31/14—Antivirals for RNA viruses
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P37/00—Drugs for immunological or allergic disorders
- A61P37/02—Immunomodulators
- A61P37/04—Immunostimulants
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/005—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from viruses
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
- C12N15/85—Vectors or expression systems specially adapted for eukaryotic hosts for animal cells
- C12N15/86—Viral vectors
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N7/00—Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/14—Hydrolases (3)
- C12N9/48—Hydrolases (3) acting on peptide bonds (3.4)
- C12N9/50—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25)
- C12N9/503—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from viruses
- C12N9/506—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from viruses derived from RNA viruses
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K2039/51—Medicinal preparations containing antigens or antibodies comprising whole cells, viruses or DNA/RNA
- A61K2039/525—Virus
- A61K2039/5254—Virus avirulent or attenuated
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K2039/51—Medicinal preparations containing antigens or antibodies comprising whole cells, viruses or DNA/RNA
- A61K2039/525—Virus
- A61K2039/5256—Virus expressing foreign proteins
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K2039/51—Medicinal preparations containing antigens or antibodies comprising whole cells, viruses or DNA/RNA
- A61K2039/53—DNA (RNA) vaccination
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K2039/55—Medicinal preparations containing antigens or antibodies characterised by the host/recipient, e.g. newborn with maternal antibodies
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2710/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA dsDNA viruses
- C12N2710/00011—Details
- C12N2710/24011—Poxviridae
- C12N2710/24111—Orthopoxvirus, e.g. vaccinia virus, variola
- C12N2710/24141—Use of virus, viral particle or viral elements as a vector
- C12N2710/24143—Use of virus, viral particle or viral elements as a vector viral genome or elements thereof as genetic vector
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2770/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssRNA viruses positive-sense
- C12N2770/00011—Details
- C12N2770/24011—Flaviviridae
- C12N2770/24211—Hepacivirus, e.g. hepatitis C virus, hepatitis G virus
- C12N2770/24222—New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2770/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssRNA viruses positive-sense
- C12N2770/00011—Details
- C12N2770/24011—Flaviviridae
- C12N2770/24211—Hepacivirus, e.g. hepatitis C virus, hepatitis G virus
- C12N2770/24234—Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
Abstract
Description
NS3/NS4AのHCV遺伝子型1a/1bコンセンサス配列の作製
NS3のコンセンサス配列を、15個の異なる遺伝子型1a配列および26個の異なる遺伝子型1b配列から作製した。NS4Aのコンセンサス配列を、15個の異なる遺伝子型1a配列および19個の異なるHCV遺伝子型1b配列から作製した。これらの配列を、標本誤差を避けるために複数の国々から選んだGenBankから入手し、Clustal X(version 1.8)ソフトウェアを用いて整列させ、最終のNS3/NS4Aコンセンサス配列を作製した。
コンセンサスNS3/NS4A配列を、IgEリーダー配列、細胞内タンパク質分解の切断部位およびC末端HAタグを付加することによりさらに改変した(図1)。GeneOptimizer(登録商標)(GENEART,Germany)を用いて、この最終配列をコドンおよびRNA最適化した。
最終のコンセンサスNS3/NS4A融合遺伝子(ConNS3/NS4A)を合成し、GENEART(Germany)によって、配列を検証した。ConNS3/NS4AをBamH1およびNot1で消化し、CMVプロモーターの制御下の臨床用発現ベクターpVAX(Invitrogen)にサブクローニングした。この最終構築物をpConNS3/NS4Aと命名した。
Lipofectamine(登録商標)(Invitrogen)を用いて、製造業者の指針に従い、Huh7.0細胞をpConNS3/NS4Aで一過的にトランスフェクトした。トランスフェクション48時間後、これらの細胞を透過処理し、この融合構築物のC末端HAタグに対するマウスモノクローナル抗体(Invitrogen)を用いて、続いて、TRITC結合ヤギ抗マウス2次抗体(Invitrogen)を用いて、タンパク質発現を調べた。
免疫化/電気穿孔
メスの6〜8週齢のC57BL/6マウスをジャクソン研究所から購入し、米国国立衛生研究所およびペンシルベニア大学の動物の施設保護および使用に関する委員会(IACUC)の指針に従って飼育した。
3回目の免疫化の1週間後にこれらのマウスを屠殺し、グループごとにそれらの脾臓を貯蔵した。Stomacher machineを用いて、これらの脾臓を粉砕し、得られた産物を40μMの細胞濾過器に通過させ、これらの脾細胞を単離した。これらの細胞を、ACK溶解緩衝液(Biosource)で5分間処理し、RBCを除去した。脾細胞の溶解後、10%FBSを加えたRPMI培地に再懸濁した。血球計を用いて、細胞数を測定した。
マウスIFN−γELISpotアッセイを、以前に記載されたように行った(Yan.,J.,et al.,Enhanced cellular immune responses elicited by an engineered HIV−1 subtype B consensus−based envelope DNA vaccine.Mol Ther,2007.15(2):p.411−21)。pConNS3/NS4Aタンパク質の8個のアミノ酸が重複し、そのタンパク質の全長に及ぶ15merのペプチドの5種類の異なるプールで、脾細胞を刺激した。これらのペプチドをインビトロジェン社が合成し、1ペプチドあたり2μg/mlの濃度で保存した。1ウェルあたり200,000細胞の濃度でこれらの脾細胞を蒔き、スポット形成単位(SFU)の平均数を、グラフ化の目的のために、1×106脾細胞に調製した。
これらの15merの重複ペプチドを21種類の別々のプールの中で貯蔵し、それぞれのペプチドはこれらの21種類のプールのうちの2つのプールに含まれる。その後、上記のIFN−γELISpotアッセイにおいて、脾細胞を各プールで刺激した。
試験設計および免疫化
実験動物の管理および使用の指針に従って、インド生まれの合計5匹のアカゲザルを使用し、飼育した。プラスミドを調製し、HPLC精製し(VGX Pharmaceuticals,Immune Therapeutics Division,The Woodlands,TX)、1%(重量/重量)ポリ−L−グルタミン酸ナトリウム塩(MW=10.5kDa)(Sigma)で調製した滅菌水で希釈した。
最初の免疫化の前に1度および各免疫化の2週間後に、これらのアカゲザルの採血を行った。これらの動物を、ケタミン(10mg/kg)およびアセプロマジン(0.1mg/kg)の混合物で麻酔した。血液をEDTA試験管に採取し、標準的なフィコール・ハイパック密度勾配遠心分離法を用いて、PBMCを単離した。単離したPBMCを完全培地(2mM/LのL−グルタミン、10%熱失活FBS、1×抗生物質/抗真菌剤、および55μM/Lのβ−メルカプトエタノールを加えたRPMI 1640)に再懸濁した。
HCV構造タンパク質NS3/NS4Aの新規HCV遺伝子型1a/1bコンセンサス融合免疫原の構築
HCVの高変異率により、様々なウイルス株に対する防御のためだけでなく、ウイルスのエスケープ変異体に対する保護による慢性感染患者における制御を維持するために、複数の免疫標的部位を有する免疫原の設計が重要である。ワクチンとの関連でコンセンサス免疫原を使用することは、天然の免疫原のみを有するワクチンと比較して、より広範な免疫反応を引き起こすことができる可能性があることを、以前の研究結果は報告している。これらの研究結果に基づき、NS3/NS4Aタンパク質に対する免疫反応の幅を拡大させることを期待して、HCV遺伝子型1a/1bのNS3/NS4Aコンセンサス配列をコードする構築物を設計した。このコンセンサス配列を、GenBankから入手した合計75種類の異なる配列から作製した。Clustal X(version 1.8)ソフトウェアを用いて、単一のコンセンサス配列を作製するのに必要とされる複数のアラインメントを作成した(図1)。
Huh7.0細胞株をpConNS3/NS4Aで一過的トランスフェクションすることにより、pConNS3/NS4Aの発現を確認した(図3)。翻訳されたタンパク質を、この構築物のC末端HAタグに対するモノクローナル抗体で検出し、免疫蛍光法を用いて視覚化した。陰性対照として、細胞を空のpVAXベクターでも一過的にトランスフェクトし、モノクローナル抗HA抗体で染色した。
pConNS3/NS4Aが、マウスにおいて細胞性免疫反応を誘導することができるかどうかを調べるために、pConNS3/NS4Aの発現の確認後、マウスをこの構築物で筋肉注射により免疫化し、続いて、電気穿孔を行った。4種類の異なる投与量のpConNS3/NS4Aを用いて、C57BL/6を3回免疫化した。これらのマウスを3回目の免疫化の1週間後に屠殺し、IFN−γELISpotアッセイを用いて、この構築物に対する細胞性免疫反応を測定した。pConNS3/NS4Aの8個のアミノ酸が重複し、その配列に及ぶ15merのペプチドの5種類のプールで、ワクチン接種したマウスの脾細胞を刺激した。図4Aに示すように、pConNS3/NS4Aは、投与量に関わらず、強い細胞性免疫反応を誘導することができる。
小動物モデルにおいては免疫原性があるが、より大きな動物に進んだ場合、一般的に、DNAワクチンは有効性を失った。しかし、pConNS3/NS4Aによって引き起こされるマウスにおける強力な細胞性免疫反応に促されたため、発明者らはより大きな動物モデルにおけるこの構築物の免疫原性を試験することを決めた。アカゲザルを、IM/EPによりpConNS3/NS4Aで、4週間の間隔を空けて、2回免疫化した(図5A)。最初の免疫化の前に1度および各免疫化の2週間後に、これらの動物の採血を行った。これらの動物のpConNS3/NS4Aに対する免疫反応をIFN−γELISpotアッセイを用いて調べた。図5Bに、3つの時点のそれぞれについて、全5種類のペプチドプールに対する各サルの反応および平均の群の反応のまとめを示す。細胞性免疫反応は、免疫化前の採血と最初の免疫化の後には検出できなかったが、2回目の免疫化後に、これらの反応は、平均555+/−280 SFU/106PBMCにまで劇的に増加した。したがって、わずか2回の免疫化後に、pConNS3/NS4Aは、確実なNS3およびNS4特異的な細胞性免疫反応を引き起こすことができた。
C57BL/6マウスにおける免疫化とは異なり、免疫反応の大部分は、単一ペプチドプールの中に含まれるpConNS3/NS4Aの1つの主要なエピトープに向けられ、免疫化されたサルの大部分(5匹中4匹)は、pConNS3/NS4Aの少なくとも2つ以上のペプチドプールに対して、強力な細胞性免疫反応を引き起こすことができた(図6)。さらなるエピトープ地図作成試験を計画するが、このことは、アカゲザルはNS3/NS4Aタンパク質内の複数の部位に対して細胞性免疫反応を引き起こすことができたことを示唆する。したがって、アカゲザルにおいて、pConNS3/NS4Aのコンセンサス配列は、NS3/NS4Aタンパク質に対する強力で、広範な細胞性免疫反応を引き起こすことができる。
急性感染から回復するHCV感染患者は、初期の、多特異性CD4+ヘルパーおよびCD8+細胞毒性T細胞反応を開始することができると知られている。HCV特異的CTL反応は、慢性感染患者におけるウイルス複製の制御に重要であることも報告された(Rehermann,B.,et al.,Quantitative analysis of the peripheral blood cytotoxic T lymphocyte response in patients with chronic hepatitis C virus infection.J Clin Invest,1996.98(6):p.1432−40;Nelson,D.R.,et al.,The role of hepatitis C virus−specific cytotoxic T lymphocytes in chronic hepatitis C.J Immunol,1997.158(3):p.1473−81)。さらに具体的には、このウイルスの構造タンパク質、特にNS3タンパク質に対する強力なT細胞反応は、急性感染のクリアランスと重要な関連があることが報告された。HCVウイルスの高変異率により、NS3などのこのウイルスの比較的保存された構造タンパク質は、T細胞に基づくワクチンに対して魅力的な候補である。
(配列表)
配列番号1
GGTACCGGATCCGCCACCATGGACTGGACCTGGATTCTGTTCCTCGTGGCTGCTGCTACAAGAGTGCACAGCGCCCCCATCACCGCCTACGCCCAGCAGACCAGGGGCCTGCTGGGCTGCATCATCACCAGCCTGACCGGCAGGGACAAGAACCAGGTGGAGGGCGAGGTGCAGGTGGTGTCCACCGCCACCCAGAGCTTTCTGGCCACCTGCATCAACGGCGTGTGCTGGACCGTGTACCATGGAGCCGGCAGCAAGACCCTGGCCGGACCCAAGGGCCCCATCACCCAGATGTACACCAACGTGGATCAGGATCTGGTCGGGTGGCCTGCCCCTCCTGGCGCCAGAAGCCTGACCCCCTGCACCTGCGGCAGCAGCGACCTGTACCTGGTGACCCGGCACGCCGACGTGATCCCCGTGCGGCGGAGAGGCGATTCCCGGGGCAGCCTGCTGTCCCCCAGGCCCATCAGCTACCTGAAGGGCAGCAGCGGCGGACCCCTGCTGTGCCCTAGCGGCCACGCCGTGGGCATCTTCAGAGCCGCCGTGTGCACCAGGGGCGTGGCCAAGGCCGTGGACTTCATCCCCGTGGAGAGCATGGAAACCACCATGCGGAGCCCCGTGTTCACCGACAACAGCAGCCCCCCAGCTGTGCCCCAGACCTTCCAGGTGGCACATCTGCACGCCCCTACCGGCAGCGGCAAGAGCACCAAGGTGCCAGCCGCCTATGCCGCCCAGGGCTACAAGGTGCTGGTGCTGAACCCCTCCGTCGCTGCTACACTGGGCTTCGGCGCCTACATGAGCAAGGCCCACGGCATCGACCCCAACATCCGGACCGGCGTGCGGACCATCACCACAGGCGCCCCTATCACATACAGCACCTACGGCAAGTTTCTGGCCGACGGCGGCTGTAGCGGCGGAGCCTACGACATCATCATCTGCGACGAGTGCCACAGCACCGACTCCACCTCCATCCTGGGCATCGGCACCGTGCTGGACCAGGCCGAGACCGCCGGAGCCAGACTGGTGGTGCTGGCCACCGCCACACCCCCTGGCAGCGTGACCGTGCCCCACCCCAATATCGAGGAAGTGGCCCTGAGCAACACCGGCGAGATCCCTTTCTACGGCAAGGCCATCCCCATCGAGGCCATCAAGGGCGGCAGGCACCTGATCTTTTGCCACAGCAAGAAGAAGTGCGACGAGCTGGCCGCCAAGCTGTCCGCCCTGGGCCTGAACGCCGTGGCCTACTACCGGGGCCTGGACGTGAGCGTGATCCCCACCTCCGGCGACGTGGTCGTGGTCGCCACAGACGCCCTGATGACCGGCTTCACCGGCGACTTCGACAGCGTGATCGACTGCAACACCTGCGTGACCCAGACCGTGGATTTCAGCCTGGACCCCACCTTCACCATCGAGACCACCACCGTGCCTCAGGACGCCGTGAGCAGAAGCCAGCGGAGGGGCCGGACCGGCAGAGGCAGGCCCGGCATCTACCGGTTCGTGACCCCTGGCGAGCGGCCCAGCGGCATGTTCGACAGCAGCGTGCTGTGCGAGTGCTACGACGCCGGCTGCGCTTGGTATGAGCTGACCCCTGCCGAGACCAGCGTGCGGCTGCGGGCCTACCTGAACACCCCAGGCCTGCCCGTGTGCCAGGACCACCTGGAATTCTGGGAGAGCGTGTTTACCGGCCTGACCCACATCGACGCCCACTTTCTGAGCCAGACCAAGCAGGCCGGCGACAACTTCCCCTACCTGGTGGCCTACCAGGCCACCGTGTGCGCCAGAGCCCAGGCCCCTCCCCCCAGCTGGGACCAGATGTGGAAGTGCCTGATCCGGCTGAAGCCCACCCTGCACGGCCCAACCCCCCTGCTGTACCGGCTGGGCGCCGTGCAGAACGAGGTGACCCTGACCCACCCTATCACCAAGTACATCATGGCCTGCATGAGCGCCGACCTGGAAGTGGTGACCAGAGGCCGGAAGCGGAGAAGCAGCACCTGGGTGCTCGTCGGCGGAGTGCTGGCTGCTCTCGCCGCCTACTGCCTGACCACCGGCAGCGTGGTGATCGTGGGCCGGATCGTGCTGTCCGGCAAGCCCGCCATCATCCCCGACCGGGAGGTGCTGTACCAGGAATTCGACGAAATGGAAGAGTGCTACCCCTACGACGTGCCCGACTACGCCTGATGAGCGGCCGCGAGTCT
配列番号2
MDWTWILFLVAAATRVHSAPITAYAQQTRGLLGCIITSLTGRDKNQVEGEVQVVSTATQSFLATCINGVCWTVYHGAGSKTLAGPKGPITQMYTNVDQDLVGWPAPPGARSLTPCTCGSSDLYLVTRHADVIPVRRRGDSRGSLLSPRPISYLKGSSGGPLLCPSGHAVGIFRAAVCTRGVAKAVDFIPVESMETTMRSPVFTDNSSPPAVPQTFQVAHLHAPTGSGKSTKVPAAYAAQGYKVLVLNPSVAATLGFGAYMSKAHGIDPNIRTGVRTITTGAPITYSTYGKFLADGGCSGGAYDIIICDECHSTDSTSILGIGTVLDQAETAGARLVVLATATPPGSVTVPHPNIEEVALSNTGEIPFYGKAIPIEAIKGGRHLIFCHSKKKCDELAAKLSALGLNAVAYYRGLDVSVIPTSGDVVVVATDALMTGFTGDFDSVIDCNTCVTQTVDFSLDPTFTIETTTVPQDAVSRSQRRGRTGRGRPGIYRFVTPGERPSGMFDSSVLCECYDAGCAWYELTPAETSVRLRAYLNTPGLPVCQDHLEFWESVFTGLTHIDAHFLSQTKQAGDNFPYLVAYQATVCARAQAPPPSWDQMWKCLIRLKPTLHGPTPLLYRLGAVQNEVTLTHPITKYIMACMSADLEVVTRGRKRRSSTWVLVGGVLAALAAYCLTTGSVVIVGRIVLSGKPAIIPDREVLYQEFDEMEECYPYDVPDYA
配列番号3
GGTACCGGATCCGCCACCATGGACTGGACCTGGATTCTGTTCCTCGTGGCTGCTGCTACAAGAGTGCACAGC
配列番号4
MDWTWILFLVAAATRVHS
配列番号5
GCCCCCATCACCGCCTACGCCCAGCAGACCAGGGGCCTGCTGGGCTGCATCATCACCAGCCTGACCGGCAGGGACAAGAACCAGGTGGAGGGCGAGGTGCAGGTGGTGTCCACCGCCACCCAGAGCTTTCTGGCCACCTGCATCAACGGCGTGTGCTGGACCGTGTACCATGGAGCCGGCAGCAAGACCCTGGCCGGACCCAAGGGCCCCATCACCCAGATGTACACCAACGTGGATCAGGATCTGGTCGGGTGGCCTGCCCCTCCTGGCGCCAGAAGCCTGACCCCCTGCACCTGCGGCAGCAGCGACCTGTACCTGGTGACCCGGCACGCCGACGTGATCCCCGTGCGGCGGAGAGGCGATTCCCGGGGCAGCCTGCTGTCCCCCAGGCCCATCAGCTACCTGAAGGGCAGCAGCGGCGGACCCCTGCTGTGCCCTAGCGGCCACGCCGTGGGCATCTTCAGAGCCGCCGTGTGCACCAGGGGCGTGGCCAAGGCCGTGGACTTCATCCCCGTGGAGAGCATGGAAACCACCATGCGGAGCCCCGTGTTCACCGACAACAGCAGCCCCCCAGCTGTGCCCCAGACCTTCCAGGTGGCACATCTGCACGCCCCTACCGGCAGCGGCAAGAGCACCAAGGTGCCAGCCGCCTATGCCGCCCAGGGCTACAAGGTGCTGGTGCTGAACCCCTCCGTCGCTGCTACACTGGGCTTCGGCGCCTACATGAGCAAGGCCCACGGCATCGACCCCAACATCCGGACCGGCGTGCGGACCATCACCACAGGCGCCCCTATCACATACAGCACCTACGGCAAGTTTCTGGCCGACGGCGGCTGTAGCGGCGGAGCCTACGACATCATCATCTGCGACGAGTGCCACAGCACCGACTCCACCTCCATCCTGGGCATCGGCACCGTGCTGGACCAGGCCGAGACCGCCGGAGCCAGACTGGTGGTGCTGGCCACCGCCACACCCCCTGGCAGCGTGACCGTGCCCCACCCCAATATCGAGGAAGTGGCCCTGAGCAACACCGGCGAGATCCCTTTCTACGGCAAGGCCATCCCCATCGAGGCCATCAAGGGCGGCAGGCACCTGATCTTTTGCCACAGCAAGAAGAAGTGCGACGAGCTGGCCGCCAAGCTGTCCGCCCTGGGCCTGAACGCCGTGGCCTACTACCGGGGCCTGGACGTGAGCGTGATCCCCACCTCCGGCGACGTGGTCGTGGTCGCCACAGACGCCCTGATGACCGGCTTCACCGGCGACTTCGACAGCGTGATCGACTGCAACACCTGCGTGACCCAGACCGTGGATTTCAGCCTGGACCCCACCTTCACCATCGAGACCACCACCGTGCCTCAGGACGCCGTGAGCAGAAGCCAGCGGAGGGGCCGGACCGGCAGAGGCAGGCCCGGCATCTACCGGTTCGTGACCCCTGGCGAGCGGCCCAGCGGCATGTTCGACAGCAGCGTGCTGTGCGAGTGCTACGACGCCGGCTGCGCTTGGTATGAGCTGACCCCTGCCGAGACCAGCGTGCGGCTGCGGGCCTACCTGAACACCCCAGGCCTGCCCGTGTGCCAGGACCACCTGGAATTCTGGGAGAGCGTGTTTACCGGCCTGACCCACATCGACGCCCACTTTCTGAGCCAGACCAAGCAGGCCGGCGACAACTTCCCCTACCTGGTGGCCTACCAGGCCACCGTGTGCGCCAGAGCCCAGGCCCCTCCCCCCAGCTGGGACCAGATGTGGAAGTGCCTGATCCGGCTGAAGCCCACCCTGCACGGCCCAACCCCCCTGCTGTACCGGCTGGGCGCCGTGCAGAACGAGGTGACCCTGACCCACCCTATCACCAAGTACATCATGGCCTGCATGAGCGCCGACCTGGAAGTGGTGACCAGAGGCCGGAAGCGGAGAAGCAGCACCTGGGTGCTCGTCGGCGGAGTGCTGGCTGCTCTCGCCGCCTACTGCCTGACCACCGGCAGCGTGGTGATCGTGGGCCGGATCGTGCTGTCCGGCAAGCCCGCCATCATCCCCGACCGGGAGGTGCTGTACCAGGAATTCGACGAAATGGAAGAGTGCTACCCCTACGACGTGCCCGACTACGCCTGATGAGCGGCCGCGAGTCT
配列番号6
APITAYAQQTRGLLGCIITSLTGRDKNQVEGEVQVVSTATQSFLATCINGVCWTVYHGAGSKTLAGPKGPITQMYTNVDQDLVGWPAPPGARSLTPCTCGSSDLYLVTRHADVIPVRRRGDSRGSLLSPRPISYLKGSSGGPLLCPSGHAVGIFRAAVCTRGVAKAVDFIPVESMETTMRSPVFTDNSSPPAVPQTFQVAHLHAPTGSGKSTKVPAAYAAQGYKVLVLNPSVAATLGFGAYMSKAHGIDPNIRTGVRTITTGAPITYSTYGKFLADGGCSGGAYDIIICDECHSTDSTSILGIGTVLDQAETAGARLVVLATATPPGSVTVPHPNIEEVALSNTGEIPFYGKAIPIEAIKGGRHLIFCHSKKKCDELAAKLSALGLNAVAYYRGLDVSVIPTSGDVVVVATDALMTGFTGDFDSVIDCNTCVTQTVDFSLDPTFTIETTTVPQDAVSRSQRRGRTGRGRPGIYRFVTPGERPSGMFDSSVLCECYDAGCAWYELTPAETSVRLRAYLNTPGLPVCQDHLEFWESVFTGLTHIDAHFLSQTKQAGDNFPYLVAYQATVCARAQAPPPSWDQMWKCLIRLKPTLHGPTPLLYRLGAVQNEVTLTHPITKYIMACMSADLEVVTRGRKRRSSTWVLVGGVLAALAAYCLTTGSVVIVGRIVLSGKPAIIPDREVLYQEFDEMEEC
配列番号7
TACCCCTACGACGTGCCCGACTACGCCTGA
配列番号8
YPYDVPDYA
配列番号9
LYRLGAVQNEVTLTH
配列番号10
GAVQNEVTH
本発明は、例えば以下の項目を提供する。
(項目1)
配列番号1;
配列番号1の断片;
配列番号1と少なくとも90%の相同性を有する配列;および
配列番号1と少なくとも90%の相同性を有する配列の断片;からなる群から選択されるヌクレオチド配列を含む核酸分子。
(項目2)
配列番号1を含む、項目1に記載の核酸分子。
(項目3)
配列番号1と少なくとも95%の相同性を有する配列を含む、項目1に記載の核酸分子。
(項目4)
配列番号1と少なくとも98%の相同性を有する配列を含む、項目1に記載の核酸分子。
(項目5)
配列番号1からなる群から選択されるヌクレオチド配列と少なくとも99%の相同性を有する配列を含む、項目1に記載の核酸分子。
(項目6)
配列番号2をコードするヌクレオチド配列;
配列番号2と少なくとも90%の相同性を有するアミノ酸配列をコードするヌクレオチド配列;
配列番号2をコードするヌクレオチド配列の断片;および
配列番号2と少なくとも90%の相同性を有するアミノ酸配列をコードするヌクレオチド配列の断片;からなる群から選択されるヌクレオチド配列を含む核酸分子。
(項目7)
配列番号2をコードするヌクレオチド配列を含む、項目6に記載の核酸分子。
(項目8)
配列番号5を含む、項目1に記載の核酸分子。
(項目9)
配列番号6をコードするヌクレオチド配列を含む、項目1に記載の核酸分子。
(項目10)
前記分子がプラスミドである、項目1〜9のいずれか一項に記載の核酸分子。
(項目11)
項目1〜10のいずれか一項に記載の核酸分子を含む医薬組成物。
(項目12)
項目1〜10のいずれか一項に記載の核酸分子を含む注射可能な医薬組成物。
(項目13)
HCVに対する免疫反応を個体において誘導する方法であって、項目1〜9のいずれか一項に記載の核酸分子を含む組成物を、前記個体に投与することを含む。
(項目14)
前記核酸分子がDNA分子である、項目13に記載の方法。
(項目15)
前記核酸分子がプラスミドである、項目14に記載の方法。
(項目16)
前記核酸分子が電気穿孔法により前記個体に導入される、項目14または15に記載の方法。
(項目17)
項目1〜9のいずれか一項に記載の核酸分子を含む組み換えワクチン。
(項目18)
前記組み換えワクチンが組み換えワクシニアワクチンである、項目17に記載の組み換えワクチン。
(項目19)
項目1〜9のいずれか一項に記載の核酸分子を含む弱毒化生ワクチン。
(項目20)
配列番号2;
配列番号2と少なくとも90%の相同性を有する配列;
配列番号2の断片;および
配列番号2と少なくとも90%の相同性を有する配列の断片;からなる群から選択されるアミノ酸配列を含むタンパク質。
(項目21)
配列番号2を含む、項目20に記載のタンパク質。
(項目22)
配列番号2と少なくとも95%の相同性を有する配列を含む、項目20に記載のタンパク質。
(項目23)
配列番号2と少なくとも98%の相同性を有する配列を含む、項目20に記載のタンパク質。
(項目24)
配列番号2と少なくとも99%の相同性を有する配列を含む、項目20に記載のタンパク質。
(項目25)
配列番号6を含む、項目20に記載のタンパク質。
(項目26)
項目20〜25のいずれか一項に記載のタンパク質を含む医薬組成物。
(項目27)
項目20〜25のいずれか一項に記載のタンパク質を含む注射可能な医薬組成物。
(項目28)
項目20〜25のいずれか一項に記載のタンパク質を含む組み換えワクチン。
(項目29)
前記組み換えワクチンが組み換えワクシニアワクチンである、項目28に記載の組み換えワクチン。
(項目30)
項目20〜25のいずれか一項に記載のタンパク質を含む弱毒化生ワクチン。
(項目31)
HCVに対する免疫反応を個体において誘導する方法であって、項目20〜25のいずれか一項に記載のタンパク質を含む組成物を、前記個体に投与することを含む方法。
(項目32)
HCVに対する免疫反応を個体において誘導する方法であって、項目17、18、28および29のいずれか一項に記載の組み換えワクチンを前記個体に投与することを含む方法。
(項目33)
HCVに対する免疫反応を個体において誘導する方法であって、項目19または30に記載の弱毒化生ワクチンを前記個体に投与することを含む方法。
(項目34)
前記個体は、HCV感染を有すると診断されている項目13〜16および31〜33のいずれか一項に記載の方法。
Claims (34)
- 配列番号1;
配列番号1の断片;
配列番号1と少なくとも90%の相同性を有する配列;および
配列番号1と少なくとも90%の相同性を有する配列の断片;からなる群から選択されるヌクレオチド配列を含む核酸分子。 - 配列番号1を含む、請求項1に記載の核酸分子。
- 配列番号1と少なくとも95%の相同性を有する配列を含む、請求項1に記載の核酸分子。
- 配列番号1と少なくとも98%の相同性を有する配列を含む、請求項1に記載の核酸分子。
- 配列番号1からなる群から選択されるヌクレオチド配列と少なくとも99%の相同性を有する配列を含む、請求項1に記載の核酸分子。
- 配列番号2をコードするヌクレオチド配列;
配列番号2と少なくとも90%の相同性を有するアミノ酸配列をコードするヌクレオチド配列;
配列番号2をコードするヌクレオチド配列の断片;および
配列番号2と少なくとも90%の相同性を有するアミノ酸配列をコードするヌクレオチド配列の断片;からなる群から選択されるヌクレオチド配列を含む核酸分子。 - 配列番号2をコードするヌクレオチド配列を含む、請求項6に記載の核酸分子。
- 配列番号5を含む、請求項1に記載の核酸分子。
- 配列番号6をコードするヌクレオチド配列を含む、請求項1に記載の核酸分子。
- 前記分子がプラスミドである、請求項1〜9のいずれか一項に記載の核酸分子。
- 請求項1〜10のいずれか一項に記載の核酸分子を含む医薬組成物。
- 請求項1〜10のいずれか一項に記載の核酸分子を含む注射可能な医薬組成物。
- HCVに対する免疫反応を個体において誘導する方法であって、請求項1〜9のいずれか一項に記載の核酸分子を含む組成物を、前記個体に投与することを含む。
- 前記核酸分子がDNA分子である、請求項13に記載の方法。
- 前記核酸分子がプラスミドである、請求項14に記載の方法。
- 前記核酸分子が電気穿孔法により前記個体に導入される、請求項14または15に記載の方法。
- 請求項1〜9のいずれか一項に記載の核酸分子を含む組み換えワクチン。
- 前記組み換えワクチンが組み換えワクシニアワクチンである、請求項17に記載の組み換えワクチン。
- 請求項1〜9のいずれか一項に記載の核酸分子を含む弱毒化生ワクチン。
- 配列番号2;
配列番号2と少なくとも90%の相同性を有する配列;
配列番号2の断片;および
配列番号2と少なくとも90%の相同性を有する配列の断片;からなる群から選択されるアミノ酸配列を含むタンパク質。 - 配列番号2を含む、請求項20に記載のタンパク質。
- 配列番号2と少なくとも95%の相同性を有する配列を含む、請求項20に記載のタンパク質。
- 配列番号2と少なくとも98%の相同性を有する配列を含む、請求項20に記載のタンパク質。
- 配列番号2と少なくとも99%の相同性を有する配列を含む、請求項20に記載のタンパク質。
- 配列番号6を含む、請求項20に記載のタンパク質。
- 請求項20〜25のいずれか一項に記載のタンパク質を含む医薬組成物。
- 請求項20〜25のいずれか一項に記載のタンパク質を含む注射可能な医薬組成物。
- 請求項20〜25のいずれか一項に記載のタンパク質を含む組み換えワクチン。
- 前記組み換えワクチンが組み換えワクシニアワクチンである、請求項28に記載の組み換えワクチン。
- 請求項20〜25のいずれか一項に記載のタンパク質を含む弱毒化生ワクチン。
- HCVに対する免疫反応を個体において誘導する方法であって、請求項20〜25のいずれか一項に記載のタンパク質を含む組成物を、前記個体に投与することを含む方法。
- HCVに対する免疫反応を個体において誘導する方法であって、請求項17、18、28および29のいずれか一項に記載の組み換えワクチンを前記個体に投与することを含む方法。
- HCVに対する免疫反応を個体において誘導する方法であって、請求項19または30に記載の弱毒化生ワクチンを前記個体に投与することを含む方法。
- 前記個体は、HCV感染を有すると診断されている請求項13〜16および31〜33のいずれか一項に記載の方法。
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
PCT/US2008/081627 WO2010050939A1 (en) | 2008-10-29 | 2008-10-29 | Improved hcv vaccines and methods for using the same |
Publications (2)
Publication Number | Publication Date |
---|---|
JP2012507280A true JP2012507280A (ja) | 2012-03-29 |
JP5753090B2 JP5753090B2 (ja) | 2015-07-22 |
Family
ID=42129096
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP2011534469A Active JP5753090B2 (ja) | 2008-10-29 | 2008-10-29 | 改良型hcvワクチンおよびその使用方法 |
Country Status (6)
Country | Link |
---|---|
US (2) | US8829174B2 (ja) |
JP (1) | JP5753090B2 (ja) |
KR (1) | KR101609233B1 (ja) |
AU (1) | AU2008363596B2 (ja) |
CA (2) | CA2674454C (ja) |
WO (1) | WO2010050939A1 (ja) |
Cited By (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2015514132A (ja) * | 2012-04-10 | 2015-05-18 | ザ トラスティーズ オブ ザ ユニバーシティ オブ ペンシルバニア | ヒト呼吸器合胞体ウイルスコンセンサス抗原、核酸構築物、およびそれらから作製されるワクチン、ならびにその使用方法 |
JP2015530410A (ja) * | 2012-09-19 | 2015-10-15 | ザ トラスティーズ オブ ザ ユニバーシティ オブ ペンシルバニア | B型肝炎ウイルスコアタンパク質および表面抗原タンパク質をコードする核酸分子ならびにそれを含むワクチン |
JP7256024B2 (ja) | 2013-03-15 | 2023-04-11 | ザ トラスティーズ オブ ザ ユニバーシティ オブ ペンシルバニア | ガンワクチン及びそれを用いた治療方法 |
Families Citing this family (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
ES2718846T3 (es) * | 2010-11-12 | 2019-07-04 | Univ Pennsylvania | Antígenos de próstata consenso, molécula de ácido nucleico que los codifica y la vacuna y usos que los comprenden |
PL2672992T3 (pl) | 2011-02-11 | 2020-11-02 | The Trustees Of The University Of Pennsylvania | Cząsteczka kwasu nukleinowego kodująca białko rdzeniowe wirusa zapalenia wątroby typu B i szczepionka je zawierająca |
CA2851336C (en) * | 2011-10-24 | 2021-01-12 | The Trustees Of The University Of Pennsylvania | Improved hcv vaccines and methods for using the same |
CN105073989A (zh) * | 2013-02-19 | 2015-11-18 | 国立大学法人神户大学 | 免疫原性多肽表层表达双歧杆菌 |
JP6376004B2 (ja) * | 2015-03-06 | 2018-08-22 | オムロン株式会社 | 無線機 |
Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2005118626A2 (en) * | 2004-06-01 | 2005-12-15 | Innogenetics N.V. | Peptides for inducing a ctl and/or htl response to hepatitis c virus |
WO2006086188A2 (en) * | 2005-01-31 | 2006-08-17 | The Johns Hopkins University | Use of consensus sequence as vaccine antigen to enhance recognition of virulent viral variants |
WO2006133911A2 (en) * | 2005-06-17 | 2006-12-21 | Istituto Di Ricerche Di Biologia Molecolare P Angeletti Spa | Hepatitis c virus nucleic acid vaccine |
WO2008107400A1 (en) * | 2007-03-02 | 2008-09-12 | Genimmune N.V. | Hcv polyepitope construct and uses thereof |
Family Cites Families (50)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US5077044A (en) | 1980-05-19 | 1991-12-31 | The Board Of Trustees Of The Leland Stanford Jr. University | Novel non-reverting shigella live vaccines |
US5110587A (en) | 1981-12-24 | 1992-05-05 | Health Research, Incorporated | Immunogenic composition comprising synthetically modified vaccinia virus |
US4722848A (en) | 1982-12-08 | 1988-02-02 | Health Research, Incorporated | Method for immunizing animals with synthetically modified vaccinia virus |
US5338683A (en) | 1981-12-24 | 1994-08-16 | Health Research Incorporated | Vaccinia virus containing DNA sequences encoding herpesvirus glycoproteins |
US5174993A (en) | 1981-12-24 | 1992-12-29 | Health Research Inc. | Recombinant avipox virus and immunological use thereof |
US4510245A (en) | 1982-11-18 | 1985-04-09 | Chiron Corporation | Adenovirus promoter system |
ZA858044B (en) | 1984-11-01 | 1987-05-27 | American Home Prod | Oral vaccines |
US5643579A (en) | 1984-11-01 | 1997-07-01 | American Home Products Corporation | Oral vaccines |
US5036006A (en) | 1984-11-13 | 1991-07-30 | Cornell Research Foundation, Inc. | Method for transporting substances into living cells and tissues and apparatus therefor |
US4945050A (en) | 1984-11-13 | 1990-07-31 | Cornell Research Foundation, Inc. | Method for transporting substances into living cells and tissues and apparatus therefor |
US4797368A (en) | 1985-03-15 | 1989-01-10 | The United States Of America As Represented By The Department Of Health And Human Services | Adeno-associated virus as eukaryotic expression vector |
GB8508845D0 (en) | 1985-04-04 | 1985-05-09 | Hoffmann La Roche | Vaccinia dna |
US5223424A (en) | 1985-09-06 | 1993-06-29 | Prutech Research And Development | Attenuated herpesviruses and herpesviruses which include foreign DNA encoding an amino acid sequence |
US4790987A (en) | 1985-11-15 | 1988-12-13 | Research Corporation | Viral glycoprotein subunit vaccine |
US5310668A (en) | 1986-06-20 | 1994-05-10 | Merck & Co., Inc. | Varicella-zoster virus as a live recombinant vaccine |
US5242829A (en) | 1986-09-23 | 1993-09-07 | Therion Biologics Corporation | Recombinant pseudorabies virus |
IL86583A0 (en) | 1987-06-04 | 1988-11-15 | Molecular Eng Ass | Vaccine containing a derivative of a microbe and method for the production thereof |
US5294441A (en) | 1987-06-04 | 1994-03-15 | Washington University | Avirulent microbes and uses therefor: salmonella typhi |
US5387744A (en) | 1987-06-04 | 1995-02-07 | Washington University | Avirulent microbes and uses therefor: Salmonella typhi |
US5112749A (en) | 1987-10-02 | 1992-05-12 | Praxis Biologics, Inc. | Vaccines for the malaria circumsporozoite protein |
CH682669A5 (fr) | 1989-03-08 | 1993-10-29 | Health Research Inc | Virus recombinant modifié pour exprimer un produit de gènes chez un hôte, procédés et vaccin correspondants. |
US5225336A (en) | 1989-03-08 | 1993-07-06 | Health Research Incorporated | Recombinant poxvirus host range selection system |
US5703055A (en) | 1989-03-21 | 1997-12-30 | Wisconsin Alumni Research Foundation | Generation of antibodies through lipid mediated DNA delivery |
US5591439A (en) | 1989-03-24 | 1997-01-07 | The Wistar Institute Of Anatomy And Biology | Recombinant cytomegalovirus vaccine |
JP3004049B2 (ja) | 1989-03-31 | 2000-01-31 | ワシントン ユニバーシティー | 病原性のないphoP型微生物を含有するワクチン |
ES2070997T3 (es) | 1989-12-04 | 1995-06-16 | Akzo Nobel Nv | Virus de herpes recombinante de pavos y vacunas vector vivas derivadas de los mismos. |
US5294548A (en) | 1990-04-02 | 1994-03-15 | American Biogenetic Sciences, Inc | Recombianant Hepatitis a virus |
WO1991018088A1 (en) | 1990-05-23 | 1991-11-28 | The United States Of America, Represented By The Secretary, United States Department Of Commerce | Adeno-associated virus (aav)-based eucaryotic vectors |
US5462734A (en) | 1990-11-02 | 1995-10-31 | Wisconsin Alumni Research Foundation | Bovine herpesvirus vaccine and method of using same |
US5240703A (en) | 1991-03-01 | 1993-08-31 | Syntro Corporation | Attenuated, genetically-engineered pseudorabies virus s-prv-155 and uses thereof |
US6034298A (en) | 1991-08-26 | 2000-03-07 | Prodigene, Inc. | Vaccines expressed in plants |
US5955088A (en) | 1992-02-03 | 1999-09-21 | Cedars-Sinai Medical Center | Pharmaceutical compsition of herpes simplex virus type-1 (HSV-1), glycoproteins |
US5981505A (en) | 1993-01-26 | 1999-11-09 | The Trustees Of The University Of Pennsylvania | Compositions and methods for delivery of genetic material |
ATE302854T1 (de) | 1993-01-26 | 2005-09-15 | Univ Pennsylvania | Zusammensetzungen und verfahren zur verabreichung von genetischem material |
US5593972A (en) | 1993-01-26 | 1997-01-14 | The Wistar Institute | Genetic immunization |
WO1994028929A1 (en) | 1993-06-07 | 1994-12-22 | Genentech, Inc. | Hiv envelope polypeptides |
US5739118A (en) | 1994-04-01 | 1998-04-14 | Apollon, Inc. | Compositions and methods for delivery of genetic material |
US6156319A (en) | 1994-07-25 | 2000-12-05 | The Trustees Of The University Of Pennsylvania | Soluble herpesvirus glycoprotein complex vaccine |
NL9401820A (nl) | 1994-11-02 | 1996-06-03 | Meyn Maschf | Inrichting voor het bewerken van aan zijn poten opgehangen gevogelte. |
US5962428A (en) | 1995-03-30 | 1999-10-05 | Apollon, Inc. | Compositions and methods for delivery of genetic material |
US5650309A (en) | 1995-05-16 | 1997-07-22 | The Regents Of The University Of California | Viral vectors |
US5698202A (en) | 1995-06-05 | 1997-12-16 | The Wistar Institute Of Anatomy & Biology | Replication-defective adenovirus human type 5 recombinant as a rabies vaccine carrier |
US6127116A (en) * | 1995-08-29 | 2000-10-03 | Washington University | Functional DNA clone for hepatitis C virus (HCV) and uses thereof |
US6589529B1 (en) | 1998-10-30 | 2003-07-08 | Children's Hospital Medical Center | Rotavirus subunit vaccine |
DE19915178A1 (de) * | 1999-04-03 | 2000-10-05 | Univ Mainz Johannes Gutenberg | Hepatitis C Virus Zellkultursystem |
JP4475561B2 (ja) | 2001-10-11 | 2010-06-09 | メルク・シャープ・エンド・ドーム・コーポレイション | C型肝炎ウイルスワクチン |
US8209006B2 (en) | 2002-03-07 | 2012-06-26 | Vgx Pharmaceuticals, Inc. | Constant current electroporation device and methods of use |
US7245963B2 (en) | 2002-03-07 | 2007-07-17 | Advisys, Inc. | Electrode assembly for constant-current electroporation and use |
WO2007031867A2 (en) | 2005-05-25 | 2007-03-22 | Tripep Ab | A hepatitis c virus non-stru tural ns3/4a fusion gene |
SG175627A1 (en) | 2006-10-17 | 2011-11-28 | Vgx Pharmaceuticals Inc | Electroporation devices and methods of using same forelectroporation of cells in mammals |
-
2008
- 2008-10-29 JP JP2011534469A patent/JP5753090B2/ja active Active
- 2008-10-29 US US13/127,008 patent/US8829174B2/en active Active
- 2008-10-29 AU AU2008363596A patent/AU2008363596B2/en active Active
- 2008-10-29 KR KR1020117010908A patent/KR101609233B1/ko active IP Right Grant
- 2008-10-29 WO PCT/US2008/081627 patent/WO2010050939A1/en active Application Filing
-
2009
- 2009-07-31 CA CA2674454A patent/CA2674454C/en active Active
- 2009-07-31 CA CA3046530A patent/CA3046530A1/en active Pending
-
2014
- 2014-09-08 US US14/480,056 patent/US9156890B2/en active Active
Patent Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2005118626A2 (en) * | 2004-06-01 | 2005-12-15 | Innogenetics N.V. | Peptides for inducing a ctl and/or htl response to hepatitis c virus |
WO2006086188A2 (en) * | 2005-01-31 | 2006-08-17 | The Johns Hopkins University | Use of consensus sequence as vaccine antigen to enhance recognition of virulent viral variants |
WO2006133911A2 (en) * | 2005-06-17 | 2006-12-21 | Istituto Di Ricerche Di Biologia Molecolare P Angeletti Spa | Hepatitis c virus nucleic acid vaccine |
WO2008107400A1 (en) * | 2007-03-02 | 2008-09-12 | Genimmune N.V. | Hcv polyepitope construct and uses thereof |
Non-Patent Citations (2)
Title |
---|
JPN6013042063; Vaccine, 2008, vol.26, p.6225-6231 [Available online 8 August 2008] * |
JPN6013042065; J. Virol., 2007, vol.81, no.15, p.8211-8224 * |
Cited By (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2015514132A (ja) * | 2012-04-10 | 2015-05-18 | ザ トラスティーズ オブ ザ ユニバーシティ オブ ペンシルバニア | ヒト呼吸器合胞体ウイルスコンセンサス抗原、核酸構築物、およびそれらから作製されるワクチン、ならびにその使用方法 |
JP2018065824A (ja) * | 2012-04-10 | 2018-04-26 | ザ トラスティーズ オブ ザ ユニバーシティ オブ ペンシルバニア | ヒト呼吸器合胞体ウイルスコンセンサス抗原、核酸構築物、およびそれらから作製されるワクチン、ならびにその使用方法 |
US10040828B2 (en) | 2012-04-10 | 2018-08-07 | The Trustees Of The University Of Pennsylvania | Human respiratory syncytial virus consensus antigens, nucleic acid constructs and vaccines made therefrom, and methods of using same |
JP2019112414A (ja) * | 2012-04-10 | 2019-07-11 | ザ トラスティーズ オブ ザ ユニバーシティ オブ ペンシルバニア | ヒト呼吸器合胞体ウイルスコンセンサス抗原、核酸構築物、およびそれらから作製されるワクチン、ならびにその使用方法 |
JP2015530410A (ja) * | 2012-09-19 | 2015-10-15 | ザ トラスティーズ オブ ザ ユニバーシティ オブ ペンシルバニア | B型肝炎ウイルスコアタンパク質および表面抗原タンパク質をコードする核酸分子ならびにそれを含むワクチン |
JP7256024B2 (ja) | 2013-03-15 | 2023-04-11 | ザ トラスティーズ オブ ザ ユニバーシティ オブ ペンシルバニア | ガンワクチン及びそれを用いた治療方法 |
Also Published As
Publication number | Publication date |
---|---|
KR101609233B1 (ko) | 2016-04-05 |
AU2008363596A1 (en) | 2010-05-06 |
JP5753090B2 (ja) | 2015-07-22 |
CA2674454A1 (en) | 2010-04-29 |
WO2010050939A1 (en) | 2010-05-06 |
AU2008363596B2 (en) | 2015-04-30 |
CA3046530A1 (en) | 2010-04-29 |
US9156890B2 (en) | 2015-10-13 |
KR20110082566A (ko) | 2011-07-19 |
US20120034256A1 (en) | 2012-02-09 |
US20150017197A1 (en) | 2015-01-15 |
US8829174B2 (en) | 2014-09-09 |
CA2674454C (en) | 2019-09-03 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
EP2234624B1 (en) | Novel vaccines against multiple subtypes of dengue virus | |
JP5753090B2 (ja) | 改良型hcvワクチンおよびその使用方法 | |
JP5744719B2 (ja) | チクングニヤウィルスタンパク質の共通配列、これをコードする核酸分子、並びにこれを使用する組成物および方法 | |
JP5996191B2 (ja) | ヒトパピローマウイルスの改良型ワクチンおよびその使用方法 | |
AU2020213308B2 (en) | Improved HCV vaccines and methods for using the same | |
US9156891B2 (en) | Vaccines and methods for using the same |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
A131 | Notification of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A131 Effective date: 20130823 |
|
A601 | Written request for extension of time |
Free format text: JAPANESE INTERMEDIATE CODE: A601 Effective date: 20131122 |
|
A602 | Written permission of extension of time |
Free format text: JAPANESE INTERMEDIATE CODE: A602 Effective date: 20131129 |
|
A601 | Written request for extension of time |
Free format text: JAPANESE INTERMEDIATE CODE: A601 Effective date: 20131220 |
|
A602 | Written permission of extension of time |
Free format text: JAPANESE INTERMEDIATE CODE: A602 Effective date: 20140106 |
|
A601 | Written request for extension of time |
Free format text: JAPANESE INTERMEDIATE CODE: A601 Effective date: 20140122 |
|
A602 | Written permission of extension of time |
Free format text: JAPANESE INTERMEDIATE CODE: A602 Effective date: 20140129 |
|
A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A821 Effective date: 20140224 |
|
A131 | Notification of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A131 Effective date: 20140904 |
|
A601 | Written request for extension of time |
Free format text: JAPANESE INTERMEDIATE CODE: A601 Effective date: 20141202 |
|
A602 | Written permission of extension of time |
Free format text: JAPANESE INTERMEDIATE CODE: A602 Effective date: 20141209 |
|
A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20141229 |
|
TRDD | Decision of grant or rejection written | ||
A01 | Written decision to grant a patent or to grant a registration (utility model) |
Free format text: JAPANESE INTERMEDIATE CODE: A01 Effective date: 20150507 |
|
A61 | First payment of annual fees (during grant procedure) |
Free format text: JAPANESE INTERMEDIATE CODE: A61 Effective date: 20150521 |
|
R150 | Certificate of patent or registration of utility model |
Ref document number: 5753090 Country of ref document: JP Free format text: JAPANESE INTERMEDIATE CODE: R150 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |