EP3445378A1 - Methods and vaccine compositions for the treatment of b-cell malignancies - Google Patents
Methods and vaccine compositions for the treatment of b-cell malignanciesInfo
- Publication number
- EP3445378A1 EP3445378A1 EP17720714.9A EP17720714A EP3445378A1 EP 3445378 A1 EP3445378 A1 EP 3445378A1 EP 17720714 A EP17720714 A EP 17720714A EP 3445378 A1 EP3445378 A1 EP 3445378A1
- Authority
- EP
- European Patent Office
- Prior art keywords
- cell
- lymphoma
- cells
- leukemia
- ebv
- 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.)
- Withdrawn
Links
- 239000000203 mixture Substances 0.000 title claims abstract description 31
- 206010028980 Neoplasm Diseases 0.000 title claims abstract description 30
- 238000000034 method Methods 0.000 title claims abstract description 30
- 238000011282 treatment Methods 0.000 title claims abstract description 22
- 229960005486 vaccine Drugs 0.000 title claims abstract description 22
- 230000036210 malignancy Effects 0.000 title claims abstract description 17
- 102100025221 CD70 antigen Human genes 0.000 claims abstract description 178
- 101000934356 Homo sapiens CD70 antigen Proteins 0.000 claims abstract description 174
- 210000003719 b-lymphocyte Anatomy 0.000 claims abstract description 137
- 241000701044 Human gammaherpesvirus 4 Species 0.000 claims abstract description 81
- 230000003211 malignant effect Effects 0.000 claims abstract description 29
- 102000039446 nucleic acids Human genes 0.000 claims abstract description 17
- 108020004707 nucleic acids Proteins 0.000 claims abstract description 17
- 150000007523 nucleic acids Chemical class 0.000 claims abstract description 17
- 201000011510 cancer Diseases 0.000 claims abstract description 15
- 230000004663 cell proliferation Effects 0.000 claims abstract description 9
- 238000012258 culturing Methods 0.000 claims abstract description 3
- 108090000623 proteins and genes Proteins 0.000 claims description 46
- 230000035772 mutation Effects 0.000 claims description 26
- 208000010839 B-cell chronic lymphocytic leukemia Diseases 0.000 claims description 19
- 206010012818 diffuse large B-cell lymphoma Diseases 0.000 claims description 18
- 206010025323 Lymphomas Diseases 0.000 claims description 16
- 208000031422 Lymphocytic Chronic B-Cell Leukemia Diseases 0.000 claims description 15
- 239000013603 viral vector Substances 0.000 claims description 13
- 208000011691 Burkitt lymphomas Diseases 0.000 claims description 12
- 208000032852 chronic lymphocytic leukemia Diseases 0.000 claims description 12
- 230000001177 retroviral effect Effects 0.000 claims description 12
- 208000017604 Hodgkin disease Diseases 0.000 claims description 10
- 208000031671 Large B-Cell Diffuse Lymphoma Diseases 0.000 claims description 10
- 208000003950 B-cell lymphoma Diseases 0.000 claims description 9
- 208000010747 Hodgkins lymphoma Diseases 0.000 claims description 9
- 206010035226 Plasma cell myeloma Diseases 0.000 claims description 9
- 208000032839 leukemia Diseases 0.000 claims description 9
- 208000015914 Non-Hodgkin lymphomas Diseases 0.000 claims description 7
- 108090000765 processed proteins & peptides Proteins 0.000 claims description 7
- 208000032568 B-cell prolymphocytic leukaemia Diseases 0.000 claims description 6
- 201000003791 MALT lymphoma Diseases 0.000 claims description 6
- 208000034578 Multiple myelomas Diseases 0.000 claims description 6
- 206010065857 Primary Effusion Lymphoma Diseases 0.000 claims description 6
- 208000035416 Prolymphocytic B-Cell Leukemia Diseases 0.000 claims description 6
- 230000001684 chronic effect Effects 0.000 claims description 6
- 201000009277 hairy cell leukemia Diseases 0.000 claims description 6
- 210000003563 lymphoid tissue Anatomy 0.000 claims description 6
- 201000007919 lymphoplasmacytic lymphoma Diseases 0.000 claims description 6
- 102000004196 processed proteins & peptides Human genes 0.000 claims description 6
- 206010062113 splenic marginal zone lymphoma Diseases 0.000 claims description 6
- 210000004877 mucosa Anatomy 0.000 claims description 5
- 229920001184 polypeptide Polymers 0.000 claims description 4
- 208000024893 Acute lymphoblastic leukemia Diseases 0.000 claims description 3
- 206010002412 Angiocentric lymphomas Diseases 0.000 claims description 3
- 208000028564 B-cell non-Hodgkin lymphoma Diseases 0.000 claims description 3
- 208000023611 Burkitt leukaemia Diseases 0.000 claims description 3
- 206010007953 Central nervous system lymphoma Diseases 0.000 claims description 3
- 208000025205 Mantle-Cell Lymphoma Diseases 0.000 claims description 3
- 206010029461 Nodal marginal zone B-cell lymphomas Diseases 0.000 claims description 3
- 206010036711 Primary mediastinal large B-cell lymphomas Diseases 0.000 claims description 3
- 208000033559 Waldenström macroglobulinemia Diseases 0.000 claims description 3
- 201000006569 extramedullary plasmacytoma Diseases 0.000 claims description 3
- 201000003444 follicular lymphoma Diseases 0.000 claims description 3
- 230000002871 immunocytoma Effects 0.000 claims description 3
- 208000026876 intravascular large B-cell lymphoma Diseases 0.000 claims description 3
- 208000006116 lymphomatoid granulomatosis Diseases 0.000 claims description 3
- 201000000050 myeloid neoplasm Diseases 0.000 claims description 3
- 208000017805 post-transplant lymphoproliferative disease Diseases 0.000 claims description 3
- 208000016800 primary central nervous system lymphoma Diseases 0.000 claims description 3
- 201000006576 solitary osseous plasmacytoma Diseases 0.000 claims description 3
- 125000003275 alpha amino acid group Chemical group 0.000 claims 2
- 210000001744 T-lymphocyte Anatomy 0.000 abstract description 138
- 230000002950 deficient Effects 0.000 abstract description 54
- 230000014509 gene expression Effects 0.000 abstract description 47
- 208000015181 infectious disease Diseases 0.000 abstract description 12
- 210000004027 cell Anatomy 0.000 description 115
- 102100027207 CD27 antigen Human genes 0.000 description 81
- 101000914511 Homo sapiens CD27 antigen Proteins 0.000 description 81
- 229940121358 tyrosine kinase inhibitor Drugs 0.000 description 53
- 102100023345 Tyrosine-protein kinase ITK/TSK Human genes 0.000 description 51
- 101001050476 Homo sapiens Tyrosine-protein kinase ITK/TSK Proteins 0.000 description 50
- 239000013598 vector Substances 0.000 description 42
- 210000003819 peripheral blood mononuclear cell Anatomy 0.000 description 33
- 229960004641 rituximab Drugs 0.000 description 22
- 229960000106 biosimilars Drugs 0.000 description 21
- 206010015108 Epstein-Barr virus infection Diseases 0.000 description 20
- 230000000638 stimulation Effects 0.000 description 20
- 210000004369 blood Anatomy 0.000 description 19
- 239000008280 blood Substances 0.000 description 19
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 18
- 230000004044 response Effects 0.000 description 18
- 102100022005 B-lymphocyte antigen CD20 Human genes 0.000 description 17
- 101000897405 Homo sapiens B-lymphocyte antigen CD20 Proteins 0.000 description 17
- 230000007812 deficiency Effects 0.000 description 17
- 230000035755 proliferation Effects 0.000 description 17
- 230000003612 virological effect Effects 0.000 description 17
- 230000005867 T cell response Effects 0.000 description 15
- 208000030289 Lymphoproliferative disease Diseases 0.000 description 14
- 235000018102 proteins Nutrition 0.000 description 14
- 102000004169 proteins and genes Human genes 0.000 description 14
- 230000004913 activation Effects 0.000 description 13
- 230000001771 impaired effect Effects 0.000 description 13
- 239000003446 ligand Substances 0.000 description 13
- 230000001404 mediated effect Effects 0.000 description 13
- 102000017420 CD3 protein, epsilon/gamma/delta subunit Human genes 0.000 description 12
- 102000003886 Glycoproteins Human genes 0.000 description 12
- 108090000288 Glycoproteins Proteins 0.000 description 12
- 102100027720 SH2 domain-containing protein 1A Human genes 0.000 description 12
- 230000001419 dependent effect Effects 0.000 description 12
- 239000003814 drug Substances 0.000 description 12
- 230000036039 immunity Effects 0.000 description 12
- 108010075704 HLA-A Antigens Proteins 0.000 description 11
- 102000011786 HLA-A Antigens Human genes 0.000 description 11
- 238000004458 analytical method Methods 0.000 description 11
- 230000006870 function Effects 0.000 description 11
- 238000012423 maintenance Methods 0.000 description 11
- 201000010099 disease Diseases 0.000 description 10
- 208000034196 Combined immunodeficiency due to ITK deficiency Diseases 0.000 description 9
- 150000001413 amino acids Chemical group 0.000 description 9
- 230000001461 cytolytic effect Effects 0.000 description 9
- 238000000684 flow cytometry Methods 0.000 description 9
- 230000006698 induction Effects 0.000 description 9
- 230000003993 interaction Effects 0.000 description 9
- 201000001280 lymphoproliferative syndrome 1 Diseases 0.000 description 9
- 230000002062 proliferating effect Effects 0.000 description 9
- 230000027425 release of sequestered calcium ion into cytosol Effects 0.000 description 9
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 8
- 102000000395 SH3 domains Human genes 0.000 description 8
- 108050008861 SH3 domains Proteins 0.000 description 8
- 108060008682 Tumor Necrosis Factor Proteins 0.000 description 8
- 241000700605 Viruses Species 0.000 description 8
- 238000005516 engineering process Methods 0.000 description 8
- 239000003623 enhancer Substances 0.000 description 8
- 230000028993 immune response Effects 0.000 description 8
- 230000037361 pathway Effects 0.000 description 8
- 230000001105 regulatory effect Effects 0.000 description 8
- 230000011664 signaling Effects 0.000 description 8
- 102000003390 tumor necrosis factor Human genes 0.000 description 8
- 108020004414 DNA Proteins 0.000 description 7
- 241000699670 Mus sp. Species 0.000 description 7
- 108010045517 Serum Amyloid P-Component Proteins 0.000 description 7
- 230000006052 T cell proliferation Effects 0.000 description 7
- 206010068348 X-linked lymphoproliferative syndrome Diseases 0.000 description 7
- 239000000427 antigen Substances 0.000 description 7
- 108091007433 antigens Proteins 0.000 description 7
- 102000036639 antigens Human genes 0.000 description 7
- 230000003185 calcium uptake Effects 0.000 description 7
- 230000001472 cytotoxic effect Effects 0.000 description 7
- 208000035475 disorder Diseases 0.000 description 7
- 239000012636 effector Substances 0.000 description 7
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 6
- 208000037435 Combined immunodeficiency due to CD27 deficiency Diseases 0.000 description 6
- 208000021519 Hodgkin lymphoma Diseases 0.000 description 6
- 239000002177 L01XE27 - Ibrutinib Substances 0.000 description 6
- 241001529936 Murinae Species 0.000 description 6
- 208000023334 autosomal recessive lymphoproliferative disease Diseases 0.000 description 6
- 239000011575 calcium Substances 0.000 description 6
- 229910052791 calcium Inorganic materials 0.000 description 6
- 238000003501 co-culture Methods 0.000 description 6
- 230000003247 decreasing effect Effects 0.000 description 6
- 238000012217 deletion Methods 0.000 description 6
- 230000037430 deletion Effects 0.000 description 6
- 229940079593 drug Drugs 0.000 description 6
- 238000002474 experimental method Methods 0.000 description 6
- 229960001507 ibrutinib Drugs 0.000 description 6
- XYFPWWZEPKGCCK-GOSISDBHSA-N ibrutinib Chemical compound C1=2C(N)=NC=NC=2N([C@H]2CN(CCC2)C(=O)C=C)N=C1C(C=C1)=CC=C1OC1=CC=CC=C1 XYFPWWZEPKGCCK-GOSISDBHSA-N 0.000 description 6
- 230000001939 inductive effect Effects 0.000 description 6
- 201000001278 lymphoproliferative syndrome 2 Diseases 0.000 description 6
- 239000013612 plasmid Substances 0.000 description 6
- UCSJYZPVAKXKNQ-HZYVHMACSA-N streptomycin Chemical compound CN[C@H]1[C@H](O)[C@@H](O)[C@H](CO)O[C@H]1O[C@@H]1[C@](C=O)(O)[C@H](C)O[C@H]1O[C@@H]1[C@@H](NC(N)=N)[C@H](O)[C@@H](NC(N)=N)[C@H](O)[C@H]1O UCSJYZPVAKXKNQ-HZYVHMACSA-N 0.000 description 6
- 238000011285 therapeutic regimen Methods 0.000 description 6
- 241001430294 unidentified retrovirus Species 0.000 description 6
- 238000007482 whole exome sequencing Methods 0.000 description 6
- 102000004127 Cytokines Human genes 0.000 description 5
- 108090000695 Cytokines Proteins 0.000 description 5
- 241000282412 Homo Species 0.000 description 5
- 241000725303 Human immunodeficiency virus Species 0.000 description 5
- 241000713666 Lentivirus Species 0.000 description 5
- 108010004729 Phycoerythrin Proteins 0.000 description 5
- 108010011033 Signaling Lymphocytic Activation Molecule Associated Protein Proteins 0.000 description 5
- 238000013459 approach Methods 0.000 description 5
- 230000027455 binding Effects 0.000 description 5
- 239000000969 carrier Substances 0.000 description 5
- 231100000433 cytotoxic Toxicity 0.000 description 5
- 210000001151 cytotoxic T lymphocyte Anatomy 0.000 description 5
- 230000007547 defect Effects 0.000 description 5
- 230000002068 genetic effect Effects 0.000 description 5
- 208000014752 hemophagocytic syndrome Diseases 0.000 description 5
- 230000001900 immune effect Effects 0.000 description 5
- 210000000265 leukocyte Anatomy 0.000 description 5
- 210000004698 lymphocyte Anatomy 0.000 description 5
- 230000007246 mechanism Effects 0.000 description 5
- 239000002609 medium Substances 0.000 description 5
- 230000000306 recurrent effect Effects 0.000 description 5
- 230000002829 reductive effect Effects 0.000 description 5
- 238000012163 sequencing technique Methods 0.000 description 5
- 238000010186 staining Methods 0.000 description 5
- 230000008685 targeting Effects 0.000 description 5
- 238000012360 testing method Methods 0.000 description 5
- 238000013518 transcription Methods 0.000 description 5
- 230000035897 transcription Effects 0.000 description 5
- 102000007469 Actins Human genes 0.000 description 4
- 108010085238 Actins Proteins 0.000 description 4
- 108010046080 CD27 Ligand Proteins 0.000 description 4
- 102100039866 CTP synthase 1 Human genes 0.000 description 4
- PMATZTZNYRCHOR-CGLBZJNRSA-N Cyclosporin A Chemical compound CC[C@@H]1NC(=O)[C@H]([C@H](O)[C@H](C)C\C=C\C)N(C)C(=O)[C@H](C(C)C)N(C)C(=O)[C@H](CC(C)C)N(C)C(=O)[C@H](CC(C)C)N(C)C(=O)[C@@H](C)NC(=O)[C@H](C)NC(=O)[C@H](CC(C)C)N(C)C(=O)[C@H](C(C)C)NC(=O)[C@H](CC(C)C)N(C)C(=O)CN(C)C1=O PMATZTZNYRCHOR-CGLBZJNRSA-N 0.000 description 4
- 108010036949 Cyclosporine Proteins 0.000 description 4
- 101001057504 Homo sapiens Interferon-stimulated gene 20 kDa protein Proteins 0.000 description 4
- 101001055144 Homo sapiens Interleukin-2 receptor subunit alpha Proteins 0.000 description 4
- 102100027268 Interferon-stimulated gene 20 kDa protein Human genes 0.000 description 4
- 102100021760 Magnesium transporter protein 1 Human genes 0.000 description 4
- 101150017238 Magt1 gene Proteins 0.000 description 4
- 241000699666 Mus <mouse, genus> Species 0.000 description 4
- 108091034117 Oligonucleotide Proteins 0.000 description 4
- 102100029215 Signaling lymphocytic activation molecule Human genes 0.000 description 4
- 101710163413 Signaling lymphocytic activation molecule Proteins 0.000 description 4
- 208000033779 X-linked lymphoproliferative disease Diseases 0.000 description 4
- 230000000259 anti-tumor effect Effects 0.000 description 4
- 238000002617 apheresis Methods 0.000 description 4
- 102220397762 c.535C>T Human genes 0.000 description 4
- 230000010261 cell growth Effects 0.000 description 4
- 239000002299 complementary DNA Substances 0.000 description 4
- 230000004940 costimulation Effects 0.000 description 4
- 230000000139 costimulatory effect Effects 0.000 description 4
- 238000004132 cross linking Methods 0.000 description 4
- 230000005860 defense response to virus Effects 0.000 description 4
- 238000010790 dilution Methods 0.000 description 4
- 239000012895 dilution Substances 0.000 description 4
- 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 4
- -1 for example Substances 0.000 description 4
- 108020001507 fusion proteins Proteins 0.000 description 4
- 239000006166 lysate Substances 0.000 description 4
- 239000002953 phosphate buffered saline Substances 0.000 description 4
- 102220271232 rs144832924 Human genes 0.000 description 4
- 102220259456 rs373042980 Human genes 0.000 description 4
- 239000000523 sample Substances 0.000 description 4
- 210000002966 serum Anatomy 0.000 description 4
- 238000002560 therapeutic procedure Methods 0.000 description 4
- 230000009466 transformation Effects 0.000 description 4
- 230000001960 triggered effect Effects 0.000 description 4
- 102000040650 (ribonucleotides)n+m Human genes 0.000 description 3
- 108700028369 Alleles Proteins 0.000 description 3
- 108091033409 CRISPR Proteins 0.000 description 3
- 238000010356 CRISPR-Cas9 genome editing Methods 0.000 description 3
- 229930105110 Cyclosporin A Natural products 0.000 description 3
- 238000001712 DNA sequencing Methods 0.000 description 3
- 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 3
- 108010074032 HLA-A2 Antigen Proteins 0.000 description 3
- 102000025850 HLA-A2 Antigen Human genes 0.000 description 3
- 101001101919 Homo sapiens CTP synthase 1 Proteins 0.000 description 3
- 101001109501 Homo sapiens NKG2-D type II integral membrane protein Proteins 0.000 description 3
- 101001136981 Homo sapiens Proteasome subunit beta type-9 Proteins 0.000 description 3
- 101000914514 Homo sapiens T-cell-specific surface glycoprotein CD28 Proteins 0.000 description 3
- 101001047681 Homo sapiens Tyrosine-protein kinase Lck Proteins 0.000 description 3
- 206010061598 Immunodeficiency Diseases 0.000 description 3
- 208000029462 Immunodeficiency disease Diseases 0.000 description 3
- 108060003951 Immunoglobulin Proteins 0.000 description 3
- 102100034349 Integrase Human genes 0.000 description 3
- 102100037850 Interferon gamma Human genes 0.000 description 3
- 108010074328 Interferon-gamma Proteins 0.000 description 3
- 108010002350 Interleukin-2 Proteins 0.000 description 3
- 241000712079 Measles morbillivirus Species 0.000 description 3
- 102100022680 NKG2-D type II integral membrane protein Human genes 0.000 description 3
- 108020004485 Nonsense Codon Proteins 0.000 description 3
- 108091028043 Nucleic acid sequence Proteins 0.000 description 3
- 241000283973 Oryctolagus cuniculus Species 0.000 description 3
- 229930182555 Penicillin Natural products 0.000 description 3
- JGSARLDLIJGVTE-MBNYWOFBSA-N Penicillin G Chemical compound N([C@H]1[C@H]2SC([C@@H](N2C1=O)C(O)=O)(C)C)C(=O)CC1=CC=CC=C1 JGSARLDLIJGVTE-MBNYWOFBSA-N 0.000 description 3
- 208000031951 Primary immunodeficiency Diseases 0.000 description 3
- 102100035764 Proteasome subunit beta type-9 Human genes 0.000 description 3
- 102000004022 Protein-Tyrosine Kinases Human genes 0.000 description 3
- 108090000412 Protein-Tyrosine Kinases Proteins 0.000 description 3
- 239000012980 RPMI-1640 medium Substances 0.000 description 3
- 241000713311 Simian immunodeficiency virus Species 0.000 description 3
- 108091027544 Subgenomic mRNA Proteins 0.000 description 3
- 102100027213 T-cell-specific surface glycoprotein CD28 Human genes 0.000 description 3
- QJJXYPPXXYFBGM-LFZNUXCKSA-N Tacrolimus Chemical compound C1C[C@@H](O)[C@H](OC)C[C@@H]1\C=C(/C)[C@@H]1[C@H](C)[C@@H](O)CC(=O)[C@H](CC=C)/C=C(C)/C[C@H](C)C[C@H](OC)[C@H]([C@H](C[C@H]2C)OC)O[C@@]2(O)C(=O)C(=O)N2CCCC[C@H]2C(=O)O1 QJJXYPPXXYFBGM-LFZNUXCKSA-N 0.000 description 3
- 108060008683 Tumor Necrosis Factor Receptor Proteins 0.000 description 3
- 102100024036 Tyrosine-protein kinase Lck Human genes 0.000 description 3
- 208000029625 X-linked lymphoproliferative disease due to SH2D1A deficiency Diseases 0.000 description 3
- 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 3
- 238000009825 accumulation Methods 0.000 description 3
- 229940024606 amino acid Drugs 0.000 description 3
- 235000001014 amino acid Nutrition 0.000 description 3
- 239000000872 buffer Substances 0.000 description 3
- 238000004113 cell culture Methods 0.000 description 3
- 238000012512 characterization method Methods 0.000 description 3
- 239000003795 chemical substances by application Substances 0.000 description 3
- 229960001265 ciclosporin Drugs 0.000 description 3
- 229930182912 cyclosporin Natural products 0.000 description 3
- 230000003013 cytotoxicity Effects 0.000 description 3
- 231100000135 cytotoxicity Toxicity 0.000 description 3
- 230000002939 deleterious effect Effects 0.000 description 3
- 210000004443 dendritic cell Anatomy 0.000 description 3
- 238000001514 detection method Methods 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 102000037865 fusion proteins Human genes 0.000 description 3
- 230000037451 immune surveillance Effects 0.000 description 3
- 230000007813 immunodeficiency Effects 0.000 description 3
- 102000018358 immunoglobulin Human genes 0.000 description 3
- 238000009169 immunotherapy Methods 0.000 description 3
- 238000001727 in vivo Methods 0.000 description 3
- PNDZEEPOYCVIIY-UHFFFAOYSA-N indo-1 Chemical compound CC1=CC=C(N(CC(O)=O)CC(O)=O)C(OCCOC=2C(=CC=C(C=2)C=2N=C3[CH]C(=CC=C3C=2)C(O)=O)N(CC(O)=O)CC(O)=O)=C1 PNDZEEPOYCVIIY-UHFFFAOYSA-N 0.000 description 3
- 238000003780 insertion Methods 0.000 description 3
- 230000037431 insertion Effects 0.000 description 3
- 230000003834 intracellular effect Effects 0.000 description 3
- 238000011835 investigation Methods 0.000 description 3
- 238000002955 isolation Methods 0.000 description 3
- 230000000670 limiting effect Effects 0.000 description 3
- 210000001165 lymph node Anatomy 0.000 description 3
- 210000001806 memory b lymphocyte Anatomy 0.000 description 3
- 230000037434 nonsense mutation Effects 0.000 description 3
- 229960002450 ofatumumab Drugs 0.000 description 3
- 238000004806 packaging method and process Methods 0.000 description 3
- 229940049954 penicillin Drugs 0.000 description 3
- 230000002085 persistent effect Effects 0.000 description 3
- 230000026731 phosphorylation Effects 0.000 description 3
- 238000006366 phosphorylation reaction Methods 0.000 description 3
- BASFCYQUMIYNBI-UHFFFAOYSA-N platinum Chemical compound [Pt] BASFCYQUMIYNBI-UHFFFAOYSA-N 0.000 description 3
- 230000037452 priming Effects 0.000 description 3
- 239000000047 product Substances 0.000 description 3
- 238000012552 review Methods 0.000 description 3
- 238000010561 standard procedure Methods 0.000 description 3
- 210000000130 stem cell Anatomy 0.000 description 3
- 229960005322 streptomycin Drugs 0.000 description 3
- 239000013589 supplement Substances 0.000 description 3
- 230000004083 survival effect Effects 0.000 description 3
- 208000024891 symptom Diseases 0.000 description 3
- QJJXYPPXXYFBGM-SHYZHZOCSA-N tacrolimus Natural products CO[C@H]1C[C@H](CC[C@@H]1O)C=C(C)[C@H]2OC(=O)[C@H]3CCCCN3C(=O)C(=O)[C@@]4(O)O[C@@H]([C@H](C[C@H]4C)OC)[C@@H](C[C@H](C)CC(=C[C@@H](CC=C)C(=O)C[C@H](O)[C@H]2C)C)OC QJJXYPPXXYFBGM-SHYZHZOCSA-N 0.000 description 3
- 230000001225 therapeutic effect Effects 0.000 description 3
- 210000004881 tumor cell Anatomy 0.000 description 3
- 102000003298 tumor necrosis factor receptor Human genes 0.000 description 3
- 238000002255 vaccination Methods 0.000 description 3
- 229950000815 veltuzumab Drugs 0.000 description 3
- YBJHBAHKTGYVGT-ZKWXMUAHSA-N (+)-Biotin Chemical compound N1C(=O)N[C@@H]2[C@H](CCCCC(=O)O)SC[C@@H]21 YBJHBAHKTGYVGT-ZKWXMUAHSA-N 0.000 description 2
- HPZMWTNATZPBIH-UHFFFAOYSA-N 1-methyladenine Chemical compound CN1C=NC2=NC=NC2=C1N HPZMWTNATZPBIH-UHFFFAOYSA-N 0.000 description 2
- RFLVMTUMFYRZCB-UHFFFAOYSA-N 1-methylguanine Chemical compound O=C1N(C)C(N)=NC2=C1N=CN2 RFLVMTUMFYRZCB-UHFFFAOYSA-N 0.000 description 2
- YSAJFXWTVFGPAX-UHFFFAOYSA-N 2-[(2,4-dioxo-1h-pyrimidin-5-yl)oxy]acetic acid Chemical compound OC(=O)COC1=CNC(=O)NC1=O YSAJFXWTVFGPAX-UHFFFAOYSA-N 0.000 description 2
- RTQWWZBSTRGEAV-PKHIMPSTSA-N 2-[[(2s)-2-[bis(carboxymethyl)amino]-3-[4-(methylcarbamoylamino)phenyl]propyl]-[2-[bis(carboxymethyl)amino]propyl]amino]acetic acid Chemical compound CNC(=O)NC1=CC=C(C[C@@H](CN(CC(C)N(CC(O)=O)CC(O)=O)CC(O)=O)N(CC(O)=O)CC(O)=O)C=C1 RTQWWZBSTRGEAV-PKHIMPSTSA-N 0.000 description 2
- FZWGECJQACGGTI-UHFFFAOYSA-N 2-amino-7-methyl-1,7-dihydro-6H-purin-6-one Chemical compound NC1=NC(O)=C2N(C)C=NC2=N1 FZWGECJQACGGTI-UHFFFAOYSA-N 0.000 description 2
- OVONXEQGWXGFJD-UHFFFAOYSA-N 4-sulfanylidene-1h-pyrimidin-2-one Chemical compound SC=1C=CNC(=O)N=1 OVONXEQGWXGFJD-UHFFFAOYSA-N 0.000 description 2
- OIVLITBTBDPEFK-UHFFFAOYSA-N 5,6-dihydrouracil Chemical compound O=C1CCNC(=O)N1 OIVLITBTBDPEFK-UHFFFAOYSA-N 0.000 description 2
- DCPSTSVLRXOYGS-UHFFFAOYSA-N 6-amino-1h-pyrimidine-2-thione Chemical compound NC1=CC=NC(S)=N1 DCPSTSVLRXOYGS-UHFFFAOYSA-N 0.000 description 2
- 241000894006 Bacteria Species 0.000 description 2
- 208000035143 Bacterial infection Diseases 0.000 description 2
- 241000283690 Bos taurus Species 0.000 description 2
- PHEDXBVPIONUQT-UHFFFAOYSA-N Cocarcinogen A1 Natural products CCCCCCCCCCCCCC(=O)OC1C(C)C2(O)C3C=C(C)C(=O)C3(O)CC(CO)=CC2C2C1(OC(C)=O)C2(C)C PHEDXBVPIONUQT-UHFFFAOYSA-N 0.000 description 2
- 108091026890 Coding region Proteins 0.000 description 2
- 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 2
- 241000713800 Feline immunodeficiency virus Species 0.000 description 2
- 238000012413 Fluorescence activated cell sorting analysis Methods 0.000 description 2
- 108010043121 Green Fluorescent Proteins Proteins 0.000 description 2
- 102000004144 Green Fluorescent Proteins Human genes 0.000 description 2
- 208000036066 Hemophagocytic Lymphohistiocytosis Diseases 0.000 description 2
- 208000032672 Histiocytosis haematophagic Diseases 0.000 description 2
- 101000601724 Homo sapiens Paired box protein Pax-5 Proteins 0.000 description 2
- 101001022129 Homo sapiens Tyrosine-protein kinase Fyn Proteins 0.000 description 2
- 206010020751 Hypersensitivity Diseases 0.000 description 2
- ONIBWKKTOPOVIA-BYPYZUCNSA-N L-Proline Chemical compound OC(=O)[C@@H]1CCCN1 ONIBWKKTOPOVIA-BYPYZUCNSA-N 0.000 description 2
- 241000829100 Macaca mulatta polyomavirus 1 Species 0.000 description 2
- 102100024193 Mitogen-activated protein kinase 1 Human genes 0.000 description 2
- 241000204031 Mycoplasma Species 0.000 description 2
- HYVABZIGRDEKCD-UHFFFAOYSA-N N(6)-dimethylallyladenine Chemical compound CC(C)=CCNC1=NC=NC2=C1N=CN2 HYVABZIGRDEKCD-UHFFFAOYSA-N 0.000 description 2
- 102100037504 Paired box protein Pax-5 Human genes 0.000 description 2
- ONIBWKKTOPOVIA-UHFFFAOYSA-N Proline Natural products OC(=O)C1CCCN1 ONIBWKKTOPOVIA-UHFFFAOYSA-N 0.000 description 2
- 102000001253 Protein Kinase Human genes 0.000 description 2
- 102000008115 Signaling Lymphocytic Activation Molecule Family Member 1 Human genes 0.000 description 2
- 108010074687 Signaling Lymphocytic Activation Molecule Family Member 1 Proteins 0.000 description 2
- 241000700584 Simplexvirus Species 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 108010090804 Streptavidin Proteins 0.000 description 2
- 229930006000 Sucrose Natural products 0.000 description 2
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 2
- 230000006044 T cell activation Effects 0.000 description 2
- 230000024932 T cell mediated immunity Effects 0.000 description 2
- 108091008874 T cell receptors Proteins 0.000 description 2
- 102000016266 T-Cell Antigen Receptors Human genes 0.000 description 2
- 108010009978 Tec protein-tyrosine kinase Proteins 0.000 description 2
- 108020000999 Viral RNA Proteins 0.000 description 2
- 230000002159 abnormal effect Effects 0.000 description 2
- 239000002253 acid Substances 0.000 description 2
- 208000026935 allergic disease Diseases 0.000 description 2
- 230000007815 allergy Effects 0.000 description 2
- 108010004469 allophycocyanin Proteins 0.000 description 2
- 230000008090 antitumoral immunity Effects 0.000 description 2
- 230000006907 apoptotic process Effects 0.000 description 2
- 238000003556 assay Methods 0.000 description 2
- 208000022362 bacterial infectious disease Diseases 0.000 description 2
- 230000033228 biological regulation Effects 0.000 description 2
- 229960000074 biopharmaceutical Drugs 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 230000037396 body weight Effects 0.000 description 2
- 210000001185 bone marrow Anatomy 0.000 description 2
- 229940112129 campath Drugs 0.000 description 2
- 230000032823 cell division Effects 0.000 description 2
- 230000007969 cellular immunity Effects 0.000 description 2
- 230000008614 cellular interaction Effects 0.000 description 2
- 230000005754 cellular signaling Effects 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 238000002512 chemotherapy Methods 0.000 description 2
- 230000001010 compromised effect Effects 0.000 description 2
- 230000016396 cytokine production Effects 0.000 description 2
- 230000007423 decrease Effects 0.000 description 2
- 230000003111 delayed effect Effects 0.000 description 2
- 239000003085 diluting agent Substances 0.000 description 2
- 230000003292 diminished effect Effects 0.000 description 2
- 230000007783 downstream signaling Effects 0.000 description 2
- 241001493065 dsRNA viruses Species 0.000 description 2
- 239000013604 expression vector Substances 0.000 description 2
- 239000012997 ficoll-paque Substances 0.000 description 2
- MHMNJMPURVTYEJ-UHFFFAOYSA-N fluorescein-5-isothiocyanate Chemical compound O1C(=O)C2=CC(N=C=S)=CC=C2C21C1=CC=C(O)C=C1OC1=CC(O)=CC=C21 MHMNJMPURVTYEJ-UHFFFAOYSA-N 0.000 description 2
- 238000001943 fluorescence-activated cell sorting Methods 0.000 description 2
- 230000009395 genetic defect Effects 0.000 description 2
- 239000005090 green fluorescent protein Substances 0.000 description 2
- 239000001963 growth medium Substances 0.000 description 2
- 229960001001 ibritumomab tiuxetan Drugs 0.000 description 2
- 210000002865 immune cell Anatomy 0.000 description 2
- 238000003119 immunoblot Methods 0.000 description 2
- 238000007901 in situ hybridization Methods 0.000 description 2
- 238000011534 incubation Methods 0.000 description 2
- 201000006747 infectious mononucleosis Diseases 0.000 description 2
- 238000001802 infusion Methods 0.000 description 2
- 239000003112 inhibitor Substances 0.000 description 2
- 230000000977 initiatory effect Effects 0.000 description 2
- 238000002347 injection Methods 0.000 description 2
- 239000007924 injection Substances 0.000 description 2
- 238000001990 intravenous administration Methods 0.000 description 2
- PGHMRUGBZOYCAA-UHFFFAOYSA-N ionomycin Natural products O1C(CC(O)C(C)C(O)C(C)C=CCC(C)CC(C)C(O)=CC(=O)C(C)CC(C)CC(CCC(O)=O)C)CCC1(C)C1OC(C)(C(C)O)CC1 PGHMRUGBZOYCAA-UHFFFAOYSA-N 0.000 description 2
- PGHMRUGBZOYCAA-ADZNBVRBSA-N ionomycin Chemical compound O1[C@H](C[C@H](O)[C@H](C)[C@H](O)[C@H](C)/C=C/C[C@@H](C)C[C@@H](C)C(/O)=C/C(=O)[C@@H](C)C[C@@H](C)C[C@@H](CCC(O)=O)C)CC[C@@]1(C)[C@@H]1O[C@](C)([C@@H](C)O)CC1 PGHMRUGBZOYCAA-ADZNBVRBSA-N 0.000 description 2
- 230000002147 killing effect Effects 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 239000003550 marker Substances 0.000 description 2
- 239000011159 matrix material Substances 0.000 description 2
- 239000012528 membrane Substances 0.000 description 2
- 230000015654 memory Effects 0.000 description 2
- MYWUZJCMWCOHBA-VIFPVBQESA-N methamphetamine Chemical compound CN[C@@H](C)CC1=CC=CC=C1 MYWUZJCMWCOHBA-VIFPVBQESA-N 0.000 description 2
- HPNSFSBZBAHARI-UHFFFAOYSA-N micophenolic acid Natural products OC1=C(CC=C(C)CCC(O)=O)C(OC)=C(C)C2=C1C(=O)OC2 HPNSFSBZBAHARI-UHFFFAOYSA-N 0.000 description 2
- 210000001616 monocyte Anatomy 0.000 description 2
- HPNSFSBZBAHARI-RUDMXATFSA-N mycophenolic acid Chemical compound OC1=C(C\C=C(/C)CCC(O)=O)C(OC)=C(C)C2=C1C(=O)OC2 HPNSFSBZBAHARI-RUDMXATFSA-N 0.000 description 2
- 210000000822 natural killer cell Anatomy 0.000 description 2
- 210000002741 palatine tonsil Anatomy 0.000 description 2
- 239000002245 particle Substances 0.000 description 2
- 244000052769 pathogen Species 0.000 description 2
- 230000001575 pathological effect Effects 0.000 description 2
- PHEDXBVPIONUQT-RGYGYFBISA-N phorbol 13-acetate 12-myristate Chemical compound C([C@]1(O)C(=O)C(C)=C[C@H]1[C@@]1(O)[C@H](C)[C@H]2OC(=O)CCCCCCCCCCCCC)C(CO)=C[C@H]1[C@H]1[C@]2(OC(C)=O)C1(C)C PHEDXBVPIONUQT-RGYGYFBISA-N 0.000 description 2
- 230000003389 potentiating effect Effects 0.000 description 2
- 230000009696 proliferative response Effects 0.000 description 2
- 229960002429 proline Drugs 0.000 description 2
- 235000013930 proline Nutrition 0.000 description 2
- 108060006633 protein kinase Proteins 0.000 description 2
- 238000010379 pull-down assay Methods 0.000 description 2
- 238000001959 radiotherapy Methods 0.000 description 2
- ZAHRKKWIAAJSAO-UHFFFAOYSA-N rapamycin Natural products COCC(O)C(=C/C(C)C(=O)CC(OC(=O)C1CCCCN1C(=O)C(=O)C2(O)OC(CC(OC)C(=CC=CC=CC(C)CC(C)C(=O)C)C)CCC2C)C(C)CC3CCC(O)C(C3)OC)C ZAHRKKWIAAJSAO-UHFFFAOYSA-N 0.000 description 2
- 102000005962 receptors Human genes 0.000 description 2
- 108020003175 receptors Proteins 0.000 description 2
- 230000010076 replication Effects 0.000 description 2
- 230000000284 resting effect Effects 0.000 description 2
- 238000003757 reverse transcription PCR Methods 0.000 description 2
- 230000002441 reversible effect Effects 0.000 description 2
- 238000013515 script Methods 0.000 description 2
- 229960002930 sirolimus Drugs 0.000 description 2
- QFJCIRLUMZQUOT-HPLJOQBZSA-N sirolimus Chemical compound C1C[C@@H](O)[C@H](OC)C[C@@H]1C[C@@H](C)[C@H]1OC(=O)[C@@H]2CCCCN2C(=O)C(=O)[C@](O)(O2)[C@H](C)CC[C@H]2C[C@H](OC)/C(C)=C/C=C/C=C/[C@@H](C)C[C@@H](C)C(=O)[C@H](OC)[C@H](O)/C(C)=C/[C@@H](C)C(=O)C1 QFJCIRLUMZQUOT-HPLJOQBZSA-N 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- 230000002269 spontaneous effect Effects 0.000 description 2
- 230000004936 stimulating effect Effects 0.000 description 2
- 239000005720 sucrose Substances 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 208000011580 syndromic disease Diseases 0.000 description 2
- 238000003786 synthesis reaction Methods 0.000 description 2
- RWQNBRDOKXIBIV-UHFFFAOYSA-N thymine Chemical compound CC1=CNC(=O)NC1=O RWQNBRDOKXIBIV-UHFFFAOYSA-N 0.000 description 2
- 210000001519 tissue Anatomy 0.000 description 2
- 238000010361 transduction Methods 0.000 description 2
- 230000026683 transduction Effects 0.000 description 2
- 230000010415 tropism Effects 0.000 description 2
- 241000701161 unidentified adenovirus Species 0.000 description 2
- 238000005406 washing Methods 0.000 description 2
- 238000001262 western blot Methods 0.000 description 2
- DGVVWUTYPXICAM-UHFFFAOYSA-N β‐Mercaptoethanol Chemical compound OCCS DGVVWUTYPXICAM-UHFFFAOYSA-N 0.000 description 2
- SATCOUWSAZBIJO-UHFFFAOYSA-N 1-methyladenine Natural products N=C1N(C)C=NC2=C1NC=N2 SATCOUWSAZBIJO-UHFFFAOYSA-N 0.000 description 1
- WJNGQIYEQLPJMN-IOSLPCCCSA-N 1-methylinosine Chemical compound C1=NC=2C(=O)N(C)C=NC=2N1[C@@H]1O[C@H](CO)[C@@H](O)[C@H]1O WJNGQIYEQLPJMN-IOSLPCCCSA-N 0.000 description 1
- HLYBTPMYFWWNJN-UHFFFAOYSA-N 2-(2,4-dioxo-1h-pyrimidin-5-yl)-2-hydroxyacetic acid Chemical compound OC(=O)C(O)C1=CNC(=O)NC1=O HLYBTPMYFWWNJN-UHFFFAOYSA-N 0.000 description 1
- SVBOROZXXYRWJL-UHFFFAOYSA-N 2-[(4-oxo-2-sulfanylidene-1h-pyrimidin-5-yl)methylamino]acetic acid Chemical compound OC(=O)CNCC1=CNC(=S)NC1=O SVBOROZXXYRWJL-UHFFFAOYSA-N 0.000 description 1
- LLWPKTDSDUQBFY-UHFFFAOYSA-N 2-[6-(aminomethyl)-2,4-dioxo-1H-pyrimidin-5-yl]acetic acid Chemical compound C(=O)(O)CC=1C(NC(NC=1CN)=O)=O LLWPKTDSDUQBFY-UHFFFAOYSA-N 0.000 description 1
- FZDFGHZZPBUTGP-UHFFFAOYSA-N 2-[[2-[bis(carboxymethyl)amino]-3-(4-isothiocyanatophenyl)propyl]-[2-[bis(carboxymethyl)amino]propyl]amino]acetic acid Chemical group OC(=O)CN(CC(O)=O)C(C)CN(CC(O)=O)CC(N(CC(O)=O)CC(O)=O)CC1=CC=C(N=C=S)C=C1 FZDFGHZZPBUTGP-UHFFFAOYSA-N 0.000 description 1
- VKWMGUNWDFIWNW-UHFFFAOYSA-N 2-chloro-1,1-dioxo-1,2-benzothiazol-3-one Chemical compound C1=CC=C2S(=O)(=O)N(Cl)C(=O)C2=C1 VKWMGUNWDFIWNW-UHFFFAOYSA-N 0.000 description 1
- XMSMHKMPBNTBOD-UHFFFAOYSA-N 2-dimethylamino-6-hydroxypurine Chemical compound N1C(N(C)C)=NC(=O)C2=C1N=CN2 XMSMHKMPBNTBOD-UHFFFAOYSA-N 0.000 description 1
- SMADWRYCYBUIKH-UHFFFAOYSA-N 2-methyl-7h-purin-6-amine Chemical compound CC1=NC(N)=C2NC=NC2=N1 SMADWRYCYBUIKH-UHFFFAOYSA-N 0.000 description 1
- KOLPWZCZXAMXKS-UHFFFAOYSA-N 3-methylcytosine Chemical compound CN1C(N)=CC=NC1=O KOLPWZCZXAMXKS-UHFFFAOYSA-N 0.000 description 1
- GJAKJCICANKRFD-UHFFFAOYSA-N 4-acetyl-4-amino-1,3-dihydropyrimidin-2-one Chemical compound CC(=O)C1(N)NC(=O)NC=C1 GJAKJCICANKRFD-UHFFFAOYSA-N 0.000 description 1
- UACOJOVKHNAJPX-UHFFFAOYSA-N 5-(methoxyamino)-6-methyl-2-sulfanylidene-1H-pyrimidin-4-one Chemical compound CONC=1C(NC(NC=1C)=S)=O UACOJOVKHNAJPX-UHFFFAOYSA-N 0.000 description 1
- MQJSSLBGAQJNER-UHFFFAOYSA-N 5-(methylaminomethyl)-1h-pyrimidine-2,4-dione Chemical compound CNCC1=CNC(=O)NC1=O MQJSSLBGAQJNER-UHFFFAOYSA-N 0.000 description 1
- LQLQRFGHAALLLE-UHFFFAOYSA-N 5-bromouracil Chemical compound BrC1=CNC(=O)NC1=O LQLQRFGHAALLLE-UHFFFAOYSA-N 0.000 description 1
- KELXHQACBIUYSE-UHFFFAOYSA-N 5-methoxy-1h-pyrimidine-2,4-dione Chemical compound COC1=CNC(=O)NC1=O KELXHQACBIUYSE-UHFFFAOYSA-N 0.000 description 1
- ZLAQATDNGLKIEV-UHFFFAOYSA-N 5-methyl-2-sulfanylidene-1h-pyrimidin-4-one Chemical compound CC1=CNC(=S)NC1=O ZLAQATDNGLKIEV-UHFFFAOYSA-N 0.000 description 1
- LRSASMSXMSNRBT-UHFFFAOYSA-N 5-methylcytosine Chemical compound CC1=CNC(=O)N=C1N LRSASMSXMSNRBT-UHFFFAOYSA-N 0.000 description 1
- HSPHKCOAUOJLIO-UHFFFAOYSA-N 6-(aziridin-1-ylamino)-1h-pyrimidin-2-one Chemical compound N1C(=O)N=CC=C1NN1CC1 HSPHKCOAUOJLIO-UHFFFAOYSA-N 0.000 description 1
- CKOMXBHMKXXTNW-UHFFFAOYSA-N 6-methyladenine Chemical compound CNC1=NC=NC2=C1N=CN2 CKOMXBHMKXXTNW-UHFFFAOYSA-N 0.000 description 1
- 108010013238 70-kDa Ribosomal Protein S6 Kinases Proteins 0.000 description 1
- SWJYOKZMYFJUOY-KQYNXXCUSA-N 9-[(2r,3r,4s,5r)-3,4-dihydroxy-5-(hydroxymethyl)oxolan-2-yl]-6-(methylamino)-7h-purin-8-one Chemical compound OC1=NC=2C(NC)=NC=NC=2N1[C@@H]1O[C@H](CO)[C@@H](O)[C@H]1O SWJYOKZMYFJUOY-KQYNXXCUSA-N 0.000 description 1
- MSSXOMSJDRHRMC-UHFFFAOYSA-N 9H-purine-2,6-diamine Chemical compound NC1=NC(N)=C2NC=NC2=N1 MSSXOMSJDRHRMC-UHFFFAOYSA-N 0.000 description 1
- 206010069754 Acquired gene mutation Diseases 0.000 description 1
- 108091093088 Amplicon Proteins 0.000 description 1
- 241000713842 Avian sarcoma virus Species 0.000 description 1
- 230000003844 B-cell-activation Effects 0.000 description 1
- 108091032955 Bacterial small RNA Proteins 0.000 description 1
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 1
- 241000701822 Bovine papillomavirus Species 0.000 description 1
- 125000001433 C-terminal amino-acid group Chemical group 0.000 description 1
- 108010029697 CD40 Ligand Proteins 0.000 description 1
- 102100032937 CD40 ligand Human genes 0.000 description 1
- 101710130444 CD70 antigen Proteins 0.000 description 1
- 101150088890 CD70 gene Proteins 0.000 description 1
- 101710155776 CTP synthase 1 Proteins 0.000 description 1
- 102000004631 Calcineurin Human genes 0.000 description 1
- 108010042955 Calcineurin Proteins 0.000 description 1
- 241000713756 Caprine arthritis encephalitis virus Species 0.000 description 1
- 108010078791 Carrier Proteins Proteins 0.000 description 1
- 231100000023 Cell-mediated cytotoxicity Toxicity 0.000 description 1
- 206010057250 Cell-mediated cytotoxicity Diseases 0.000 description 1
- 206010010099 Combined immunodeficiency Diseases 0.000 description 1
- 108020004635 Complementary DNA Proteins 0.000 description 1
- 108091035707 Consensus sequence Proteins 0.000 description 1
- 241000557626 Corvus corax Species 0.000 description 1
- CMSMOCZEIVJLDB-UHFFFAOYSA-N Cyclophosphamide Chemical compound ClCCN(CCCl)P1(=O)NCCCO1 CMSMOCZEIVJLDB-UHFFFAOYSA-N 0.000 description 1
- 241000701022 Cytomegalovirus Species 0.000 description 1
- 206010011831 Cytomegalovirus infection Diseases 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
- WQZGKKKJIJFFOK-QTVWNMPRSA-N D-mannopyranose Chemical compound OC[C@H]1OC(O)[C@@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-QTVWNMPRSA-N 0.000 description 1
- 239000003155 DNA primer Substances 0.000 description 1
- 102000004163 DNA-directed RNA polymerases Human genes 0.000 description 1
- 108090000626 DNA-directed RNA polymerases Proteins 0.000 description 1
- 108010008532 Deoxyribonuclease I Proteins 0.000 description 1
- 102000007260 Deoxyribonuclease I Human genes 0.000 description 1
- 241000702421 Dependoparvovirus Species 0.000 description 1
- 229920002307 Dextran Polymers 0.000 description 1
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 1
- 102100035102 E3 ubiquitin-protein ligase MYCBP2 Human genes 0.000 description 1
- 101710121417 Envelope glycoprotein Proteins 0.000 description 1
- 101710091045 Envelope protein Proteins 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- BRLQWZUYTZBJKN-UHFFFAOYSA-N Epichlorohydrin Chemical compound ClCC1CO1 BRLQWZUYTZBJKN-UHFFFAOYSA-N 0.000 description 1
- 208000016393 Epstein-Barr virus-associated malignant lymphoproliferative disease Diseases 0.000 description 1
- 241000713730 Equine infectious anemia virus Species 0.000 description 1
- 108700024394 Exon Proteins 0.000 description 1
- 229940126611 FBTA05 Drugs 0.000 description 1
- 241000700662 Fowlpox virus Species 0.000 description 1
- 108700028146 Genetic Enhancer Elements Proteins 0.000 description 1
- 101001093025 Geobacillus stearothermophilus 50S ribosomal protein L7/L12 Proteins 0.000 description 1
- 208000032612 Glial tumor Diseases 0.000 description 1
- 206010018338 Glioma Diseases 0.000 description 1
- 239000012571 GlutaMAX medium Substances 0.000 description 1
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 description 1
- 108010035452 HLA-A1 Antigen Proteins 0.000 description 1
- 101710154606 Hemagglutinin Proteins 0.000 description 1
- 208000014669 Hemophagocytic syndrome associated with an infection Diseases 0.000 description 1
- 241000700721 Hepatitis B virus Species 0.000 description 1
- 101100503640 Homo sapiens FYN gene Proteins 0.000 description 1
- 101001023379 Homo sapiens Lysosome-associated membrane glycoprotein 1 Proteins 0.000 description 1
- 101000638251 Homo sapiens Tumor necrosis factor ligand superfamily member 9 Proteins 0.000 description 1
- 241000713340 Human immunodeficiency virus 2 Species 0.000 description 1
- 229930010555 Inosine Natural products 0.000 description 1
- UGQMRVRMYYASKQ-KQYNXXCUSA-N Inosine Chemical compound O[C@@H]1[C@H](O)[C@@H](CO)O[C@H]1N1C2=NC=NC(O)=C2N=C1 UGQMRVRMYYASKQ-KQYNXXCUSA-N 0.000 description 1
- 108090001005 Interleukin-6 Proteins 0.000 description 1
- 108010038498 Interleukin-7 Receptors Proteins 0.000 description 1
- 102100021593 Interleukin-7 receptor subunit alpha Human genes 0.000 description 1
- 108091092195 Intron Proteins 0.000 description 1
- 101150025881 Itk gene Proteins 0.000 description 1
- 108010055717 JNK Mitogen-Activated Protein Kinases Proteins 0.000 description 1
- PWKSKIMOESPYIA-BYPYZUCNSA-N L-N-acetyl-Cysteine Chemical compound CC(=O)N[C@@H](CS)C(O)=O PWKSKIMOESPYIA-BYPYZUCNSA-N 0.000 description 1
- FBOZXECLQNJBKD-ZDUSSCGKSA-N L-methotrexate Chemical compound C=1N=C2N=C(N)N=C(N)C2=NC=1CN(C)C1=CC=C(C(=O)N[C@@H](CCC(O)=O)C(O)=O)C=C1 FBOZXECLQNJBKD-ZDUSSCGKSA-N 0.000 description 1
- OUYCCCASQSFEME-QMMMGPOBSA-N L-tyrosine Chemical compound OC(=O)[C@@H](N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-QMMMGPOBSA-N 0.000 description 1
- 208000008771 Lymphadenopathy Diseases 0.000 description 1
- 241000712899 Lymphocytic choriomeningitis mammarenavirus Species 0.000 description 1
- 102100035133 Lysosome-associated membrane glycoprotein 1 Human genes 0.000 description 1
- 102000019149 MAP kinase activity proteins Human genes 0.000 description 1
- 108040008097 MAP kinase activity proteins Proteins 0.000 description 1
- FYYHWMGAXLPEAU-UHFFFAOYSA-N Magnesium Chemical compound [Mg] FYYHWMGAXLPEAU-UHFFFAOYSA-N 0.000 description 1
- 241000124008 Mammalia Species 0.000 description 1
- 229930195725 Mannitol Natural products 0.000 description 1
- 108010052285 Membrane Proteins Proteins 0.000 description 1
- 206010027476 Metastases Diseases 0.000 description 1
- 241000713869 Moloney murine leukemia virus Species 0.000 description 1
- 241000714177 Murine leukemia virus Species 0.000 description 1
- 101500027988 Mus musculus ADGRV1 subunit beta Proteins 0.000 description 1
- 101100519207 Mus musculus Pdcd1 gene Proteins 0.000 description 1
- 108010021466 Mutant Proteins Proteins 0.000 description 1
- 102000008300 Mutant Proteins Human genes 0.000 description 1
- 102000004364 Myogenin Human genes 0.000 description 1
- 108010056785 Myogenin Proteins 0.000 description 1
- SGSSKEDGVONRGC-UHFFFAOYSA-N N(2)-methylguanine Chemical compound O=C1NC(NC)=NC2=C1N=CN2 SGSSKEDGVONRGC-UHFFFAOYSA-N 0.000 description 1
- 108010057466 NF-kappa B Proteins 0.000 description 1
- 108010014632 NF-kappa B kinase Proteins 0.000 description 1
- 102000019148 NF-kappaB-inducing kinase activity proteins Human genes 0.000 description 1
- 102100023050 Nuclear factor NF-kappa-B p105 subunit Human genes 0.000 description 1
- 101710163270 Nuclease Proteins 0.000 description 1
- 101710093908 Outer capsid protein VP4 Proteins 0.000 description 1
- 101710135467 Outer capsid protein sigma-1 Proteins 0.000 description 1
- 238000012408 PCR amplification Methods 0.000 description 1
- 239000002033 PVDF binder Substances 0.000 description 1
- 241001505332 Polyomavirus sp. Species 0.000 description 1
- 102100037935 Polyubiquitin-C Human genes 0.000 description 1
- 229940124158 Protease/peptidase inhibitor Drugs 0.000 description 1
- 101710176177 Protein A56 Proteins 0.000 description 1
- 101710188315 Protein X Proteins 0.000 description 1
- 238000012193 PureLink RNA Mini Kit Methods 0.000 description 1
- 206010037660 Pyrexia Diseases 0.000 description 1
- 241001510071 Pyrrhocoridae Species 0.000 description 1
- 238000012181 QIAquick gel extraction kit Methods 0.000 description 1
- 230000004570 RNA-binding Effects 0.000 description 1
- 108010092799 RNA-directed DNA polymerase Proteins 0.000 description 1
- 238000011529 RT qPCR Methods 0.000 description 1
- 208000037340 Rare genetic disease Diseases 0.000 description 1
- 206010057190 Respiratory tract infections Diseases 0.000 description 1
- 241000283984 Rodentia Species 0.000 description 1
- 239000006146 Roswell Park Memorial Institute medium Substances 0.000 description 1
- 102000014400 SH2 domains Human genes 0.000 description 1
- 108050003452 SH2 domains Proteins 0.000 description 1
- 108700015968 Slam family Proteins 0.000 description 1
- 108091081024 Start codon Proteins 0.000 description 1
- 108700025695 Suppressor Genes Proteins 0.000 description 1
- 230000020385 T cell costimulation Effects 0.000 description 1
- 108090000925 TNF receptor-associated factor 2 Proteins 0.000 description 1
- 102000003718 TNF receptor-associated factor 5 Human genes 0.000 description 1
- 108090000001 TNF receptor-associated factor 5 Proteins 0.000 description 1
- 102000002259 TNF-Related Apoptosis-Inducing Ligand Receptors Human genes 0.000 description 1
- 108010000449 TNF-Related Apoptosis-Inducing Ligand Receptors Proteins 0.000 description 1
- 102100034779 TRAF family member-associated NF-kappa-B activator Human genes 0.000 description 1
- 108700042805 TRU-015 Proteins 0.000 description 1
- 108010006785 Taq Polymerase Proteins 0.000 description 1
- 239000004098 Tetracycline Substances 0.000 description 1
- 108700019146 Transgenes Proteins 0.000 description 1
- 101800001690 Transmembrane protein gp41 Proteins 0.000 description 1
- 239000007983 Tris buffer Substances 0.000 description 1
- 102100022153 Tumor necrosis factor receptor superfamily member 4 Human genes 0.000 description 1
- 101710165473 Tumor necrosis factor receptor superfamily member 4 Proteins 0.000 description 1
- 108091005956 Type II transmembrane proteins Proteins 0.000 description 1
- 229940127174 UCHT1 Drugs 0.000 description 1
- 108090000848 Ubiquitin Proteins 0.000 description 1
- 102000044159 Ubiquitin Human genes 0.000 description 1
- 108010056354 Ubiquitin C Proteins 0.000 description 1
- 102000005918 Ubiquitin Thiolesterase Human genes 0.000 description 1
- 108010005656 Ubiquitin Thiolesterase Proteins 0.000 description 1
- 241000700618 Vaccinia virus Species 0.000 description 1
- 208000010094 Visna Diseases 0.000 description 1
- 208000026309 X-linked immunodeficiency with magnesium defect, Epstein-Barr virus infection and neoplasia Diseases 0.000 description 1
- 201000006722 X-linked immunodeficiency with magnesium defect, Epstein-Barr virus infection, and neoplasia Diseases 0.000 description 1
- 238000010317 ablation therapy Methods 0.000 description 1
- 230000005856 abnormality Effects 0.000 description 1
- 229960004308 acetylcysteine Drugs 0.000 description 1
- CAWBRCOBJNWRLK-UHFFFAOYSA-N acetyloxymethyl 2-[4-[bis[2-(acetyloxymethoxy)-2-oxoethyl]amino]-3-[2-[2-[bis[2-(acetyloxymethoxy)-2-oxoethyl]amino]-5-methylphenoxy]ethoxy]phenyl]-1h-indole-6-carboxylate Chemical compound CC(=O)OCOC(=O)CN(CC(=O)OCOC(C)=O)C1=CC=C(C)C=C1OCCOC1=CC(C=2NC3=CC(=CC=C3C=2)C(=O)OCOC(C)=O)=CC=C1N(CC(=O)OCOC(C)=O)CC(=O)OCOC(C)=O CAWBRCOBJNWRLK-UHFFFAOYSA-N 0.000 description 1
- 150000007513 acids Chemical class 0.000 description 1
- 230000003213 activating effect Effects 0.000 description 1
- 230000006786 activation induced cell death Effects 0.000 description 1
- 230000001154 acute effect Effects 0.000 description 1
- 230000008649 adaptation response Effects 0.000 description 1
- 101150063416 add gene Proteins 0.000 description 1
- 239000002671 adjuvant Substances 0.000 description 1
- 239000000443 aerosol Substances 0.000 description 1
- 230000002776 aggregation Effects 0.000 description 1
- 238000004220 aggregation Methods 0.000 description 1
- 230000000735 allogeneic effect Effects 0.000 description 1
- WNROFYMDJYEPJX-UHFFFAOYSA-K aluminium hydroxide Chemical compound [OH-].[OH-].[OH-].[Al+3] WNROFYMDJYEPJX-UHFFFAOYSA-K 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
- 210000004102 animal cell Anatomy 0.000 description 1
- 239000003242 anti bacterial agent Substances 0.000 description 1
- 230000001348 anti-glioma Effects 0.000 description 1
- 230000005809 anti-tumor immunity Effects 0.000 description 1
- 229940088710 antibiotic agent Drugs 0.000 description 1
- 238000009175 antibody therapy Methods 0.000 description 1
- 230000014102 antigen processing and presentation of exogenous peptide antigen via MHC class I Effects 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
- 239000003443 antiviral agent Substances 0.000 description 1
- 230000002238 attenuated effect Effects 0.000 description 1
- LMEKQMALGUDUQG-UHFFFAOYSA-N azathioprine Chemical compound CN1C=NC([N+]([O-])=O)=C1SC1=NC=NC2=C1NC=N2 LMEKQMALGUDUQG-UHFFFAOYSA-N 0.000 description 1
- 229960002170 azathioprine Drugs 0.000 description 1
- 210000000649 b-lymphocyte subset Anatomy 0.000 description 1
- 239000011324 bead Substances 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
- 238000001574 biopsy Methods 0.000 description 1
- 229940126587 biotherapeutics Drugs 0.000 description 1
- 229960002685 biotin Drugs 0.000 description 1
- 235000020958 biotin Nutrition 0.000 description 1
- 239000011616 biotin Substances 0.000 description 1
- 210000001772 blood platelet Anatomy 0.000 description 1
- 238000010322 bone marrow transplantation Methods 0.000 description 1
- KQNZDYYTLMIZCT-KQPMLPITSA-N brefeldin A Chemical compound O[C@@H]1\C=C\C(=O)O[C@@H](C)CCC\C=C\[C@@H]2C[C@H](O)C[C@H]21 KQNZDYYTLMIZCT-KQPMLPITSA-N 0.000 description 1
- JUMGSHROWPPKFX-UHFFFAOYSA-N brefeldin-A Natural products CC1CCCC=CC2(C)CC(O)CC2(C)C(O)C=CC(=O)O1 JUMGSHROWPPKFX-UHFFFAOYSA-N 0.000 description 1
- 239000007975 buffered saline Substances 0.000 description 1
- 230000001275 ca(2+)-mobilization Effects 0.000 description 1
- 238000004422 calculation algorithm Methods 0.000 description 1
- 238000004364 calculation method Methods 0.000 description 1
- 238000002619 cancer immunotherapy Methods 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
- 235000014633 carbohydrates Nutrition 0.000 description 1
- 150000001768 cations Chemical class 0.000 description 1
- 230000030833 cell death Effects 0.000 description 1
- 239000013553 cell monolayer Substances 0.000 description 1
- 238000001516 cell proliferation assay Methods 0.000 description 1
- 230000005890 cell-mediated cytotoxicity Effects 0.000 description 1
- 230000036755 cellular response Effects 0.000 description 1
- 239000002738 chelating agent Substances 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 239000003638 chemical reducing agent Substances 0.000 description 1
- 210000000349 chromosome Anatomy 0.000 description 1
- 208000013056 classic Hodgkin lymphoma Diseases 0.000 description 1
- 238000010367 cloning Methods 0.000 description 1
- 238000011109 contamination Methods 0.000 description 1
- 238000011340 continuous therapy Methods 0.000 description 1
- 229920001577 copolymer Polymers 0.000 description 1
- 230000008878 coupling Effects 0.000 description 1
- 238000010168 coupling process Methods 0.000 description 1
- 238000005859 coupling reaction Methods 0.000 description 1
- 239000013078 crystal Substances 0.000 description 1
- 229960004397 cyclophosphamide Drugs 0.000 description 1
- 230000009089 cytolysis Effects 0.000 description 1
- 230000007402 cytotoxic response Effects 0.000 description 1
- 238000002784 cytotoxicity assay Methods 0.000 description 1
- 230000034994 death Effects 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000029087 digestion Effects 0.000 description 1
- 239000000539 dimer Substances 0.000 description 1
- 230000005014 ectopic expression Effects 0.000 description 1
- 210000003162 effector t lymphocyte Anatomy 0.000 description 1
- 238000004520 electroporation Methods 0.000 description 1
- 108010075324 emt protein-tyrosine kinase Proteins 0.000 description 1
- 108700004025 env Genes Proteins 0.000 description 1
- 101150030339 env gene Proteins 0.000 description 1
- 244000309457 enveloped RNA virus Species 0.000 description 1
- 210000003743 erythrocyte Anatomy 0.000 description 1
- 210000003527 eukaryotic cell Anatomy 0.000 description 1
- 230000028023 exocytosis Effects 0.000 description 1
- 238000010195 expression analysis Methods 0.000 description 1
- 238000002710 external beam radiation therapy Methods 0.000 description 1
- 210000004700 fetal blood Anatomy 0.000 description 1
- 239000012894 fetal calf serum Substances 0.000 description 1
- 230000001605 fetal effect Effects 0.000 description 1
- 230000037406 food intake Effects 0.000 description 1
- 239000012634 fragment Substances 0.000 description 1
- 108700004026 gag Genes Proteins 0.000 description 1
- 101150098622 gag gene Proteins 0.000 description 1
- 229930192878 garvin Natural products 0.000 description 1
- 238000001415 gene therapy Methods 0.000 description 1
- 238000010353 genetic engineering Methods 0.000 description 1
- 102000054766 genetic haplotypes Human genes 0.000 description 1
- 230000007614 genetic variation Effects 0.000 description 1
- 238000003205 genotyping method Methods 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- RWSXRVCMGQZWBV-WDSKDSINSA-N glutathione Chemical compound OC(=O)[C@@H](N)CCC(=O)N[C@@H](CS)C(=O)NCC(O)=O RWSXRVCMGQZWBV-WDSKDSINSA-N 0.000 description 1
- 230000013595 glycosylation Effects 0.000 description 1
- 238000006206 glycosylation reaction Methods 0.000 description 1
- 108010065889 glycyl-leucyl-cysteinyl-threonyl-leucyl-valyl-alanyl-methionyl-leucine Proteins 0.000 description 1
- 239000008187 granular material Substances 0.000 description 1
- 210000003714 granulocyte Anatomy 0.000 description 1
- 239000003102 growth factor Substances 0.000 description 1
- 230000003394 haemopoietic effect Effects 0.000 description 1
- 239000000185 hemagglutinin Substances 0.000 description 1
- 210000003958 hematopoietic stem cell Anatomy 0.000 description 1
- 238000011134 hematopoietic stem cell transplantation Methods 0.000 description 1
- 206010019847 hepatosplenomegaly Diseases 0.000 description 1
- 239000000710 homodimer Substances 0.000 description 1
- 102000053826 human CD70 Human genes 0.000 description 1
- 102000045725 human FYN Human genes 0.000 description 1
- 229940072221 immunoglobulins Drugs 0.000 description 1
- 239000003018 immunosuppressive agent Substances 0.000 description 1
- 229940125721 immunosuppressive agent Drugs 0.000 description 1
- 238000002513 implantation Methods 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 230000002779 inactivation Effects 0.000 description 1
- 238000010348 incorporation Methods 0.000 description 1
- 206010022000 influenza Diseases 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 238000011221 initial treatment Methods 0.000 description 1
- 229960003786 inosine Drugs 0.000 description 1
- 238000010212 intracellular staining Methods 0.000 description 1
- 238000010253 intravenous injection Methods 0.000 description 1
- 229940036646 iodine-131-tositumomab Drugs 0.000 description 1
- 230000002427 irreversible effect Effects 0.000 description 1
- 238000002372 labelling Methods 0.000 description 1
- 238000011068 loading method Methods 0.000 description 1
- 230000005923 long-lasting effect Effects 0.000 description 1
- 230000007787 long-term memory Effects 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 230000004777 loss-of-function mutation Effects 0.000 description 1
- 210000004324 lymphatic system Anatomy 0.000 description 1
- 208000019420 lymphoid neoplasm Diseases 0.000 description 1
- 230000001589 lymphoproliferative effect Effects 0.000 description 1
- 230000002101 lytic effect Effects 0.000 description 1
- 239000011777 magnesium Substances 0.000 description 1
- 229910052749 magnesium Inorganic materials 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
- 230000008774 maternal effect Effects 0.000 description 1
- 230000010534 mechanism of action Effects 0.000 description 1
- 210000003071 memory t lymphocyte Anatomy 0.000 description 1
- 108020004999 messenger RNA Proteins 0.000 description 1
- 230000009401 metastasis Effects 0.000 description 1
- 229960000485 methotrexate Drugs 0.000 description 1
- IZAGSTRIDUNNOY-UHFFFAOYSA-N methyl 2-[(2,4-dioxo-1h-pyrimidin-5-yl)oxy]acetate Chemical compound COC(=O)COC1=CNC(=O)NC1=O IZAGSTRIDUNNOY-UHFFFAOYSA-N 0.000 description 1
- 150000004702 methyl esters Chemical class 0.000 description 1
- 229950003734 milatuzumab Drugs 0.000 description 1
- 235000013336 milk Nutrition 0.000 description 1
- 239000008267 milk Substances 0.000 description 1
- 210000004080 milk Anatomy 0.000 description 1
- 238000010369 molecular cloning Methods 0.000 description 1
- 238000003032 molecular docking Methods 0.000 description 1
- 230000009456 molecular mechanism Effects 0.000 description 1
- 230000004784 molecular pathogenesis Effects 0.000 description 1
- 239000003068 molecular probe Substances 0.000 description 1
- 238000012544 monitoring process Methods 0.000 description 1
- 238000010172 mouse model Methods 0.000 description 1
- 229940014456 mycophenolate Drugs 0.000 description 1
- 229960000951 mycophenolic acid Drugs 0.000 description 1
- XJVXMWNLQRTRGH-UHFFFAOYSA-N n-(3-methylbut-3-enyl)-2-methylsulfanyl-7h-purin-6-amine Chemical compound CSC1=NC(NCCC(C)=C)=C2NC=NC2=N1 XJVXMWNLQRTRGH-UHFFFAOYSA-N 0.000 description 1
- 230000009826 neoplastic cell growth Effects 0.000 description 1
- 230000007935 neutral effect Effects 0.000 description 1
- 210000000440 neutrophil Anatomy 0.000 description 1
- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- 239000002773 nucleotide Substances 0.000 description 1
- 125000003729 nucleotide group Chemical group 0.000 description 1
- 229960003347 obinutuzumab Drugs 0.000 description 1
- 229950009090 ocaratuzumab Drugs 0.000 description 1
- 229950005751 ocrelizumab Drugs 0.000 description 1
- 230000036961 partial effect Effects 0.000 description 1
- 230000008506 pathogenesis Effects 0.000 description 1
- 230000007310 pathophysiology Effects 0.000 description 1
- 239000000137 peptide hydrolase inhibitor Substances 0.000 description 1
- 210000005259 peripheral blood Anatomy 0.000 description 1
- 239000011886 peripheral blood Substances 0.000 description 1
- 210000004976 peripheral blood cell Anatomy 0.000 description 1
- 210000005105 peripheral blood lymphocyte Anatomy 0.000 description 1
- 239000000546 pharmaceutical excipient Substances 0.000 description 1
- 108010058237 plasma protein fraction Proteins 0.000 description 1
- 229940002993 plasmanate Drugs 0.000 description 1
- 210000004180 plasmocyte Anatomy 0.000 description 1
- 229910052697 platinum Inorganic materials 0.000 description 1
- 108700004029 pol Genes Proteins 0.000 description 1
- 101150088264 pol gene Proteins 0.000 description 1
- 230000008488 polyadenylation Effects 0.000 description 1
- 238000003752 polymerase chain reaction Methods 0.000 description 1
- 108091033319 polynucleotide Proteins 0.000 description 1
- 102000040430 polynucleotide Human genes 0.000 description 1
- 239000002157 polynucleotide Substances 0.000 description 1
- 229920002981 polyvinylidene fluoride Polymers 0.000 description 1
- 239000002243 precursor Substances 0.000 description 1
- 239000003755 preservative agent Substances 0.000 description 1
- 230000003449 preventive effect Effects 0.000 description 1
- 239000013615 primer Substances 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- ZYHMJXZULPZUED-UHFFFAOYSA-N propargite Chemical compound C1=CC(C(C)(C)C)=CC=C1OC1C(OS(=O)OCC#C)CCCC1 ZYHMJXZULPZUED-UHFFFAOYSA-N 0.000 description 1
- 230000000069 prophylactic effect Effects 0.000 description 1
- 238000000159 protein binding assay Methods 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 230000005855 radiation Effects 0.000 description 1
- 230000002285 radioactive effect Effects 0.000 description 1
- 238000003753 real-time PCR Methods 0.000 description 1
- 210000001350 reed-sternberg cell Anatomy 0.000 description 1
- 108091008146 restriction endonucleases Proteins 0.000 description 1
- 238000010839 reverse transcription Methods 0.000 description 1
- 229960003452 romidepsin Drugs 0.000 description 1
- OHRURASPPZQGQM-GCCNXGTGSA-N romidepsin Chemical compound O1C(=O)[C@H](C(C)C)NC(=O)C(=C/C)/NC(=O)[C@H]2CSSCC\C=C\[C@@H]1CC(=O)N[C@H](C(C)C)C(=O)N2 OHRURASPPZQGQM-GCCNXGTGSA-N 0.000 description 1
- 102200107843 rs11540654 Human genes 0.000 description 1
- 102220023060 rs397514261 Human genes 0.000 description 1
- 238000007480 sanger sequencing Methods 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 238000002864 sequence alignment Methods 0.000 description 1
- 230000019491 signal transduction Effects 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 1
- YEENEYXBHNNNGV-XEHWZWQGSA-M sodium;3-acetamido-5-[acetyl(methyl)amino]-2,4,6-triiodobenzoate;(2r,3r,4s,5s,6r)-2-[(2r,3s,4s,5r)-3,4-dihydroxy-2,5-bis(hydroxymethyl)oxolan-2-yl]oxy-6-(hydroxymethyl)oxane-3,4,5-triol Chemical compound [Na+].CC(=O)N(C)C1=C(I)C(NC(C)=O)=C(I)C(C([O-])=O)=C1I.O[C@H]1[C@H](O)[C@@H](CO)O[C@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 YEENEYXBHNNNGV-XEHWZWQGSA-M 0.000 description 1
- 230000037439 somatic mutation Effects 0.000 description 1
- 210000000952 spleen Anatomy 0.000 description 1
- 230000010473 stable expression Effects 0.000 description 1
- 150000003431 steroids Chemical class 0.000 description 1
- 238000010254 subcutaneous injection Methods 0.000 description 1
- 239000007929 subcutaneous injection Substances 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 230000008093 supporting effect Effects 0.000 description 1
- 230000003319 supportive effect Effects 0.000 description 1
- 230000001629 suppression Effects 0.000 description 1
- 239000004094 surface-active agent Substances 0.000 description 1
- 230000002459 sustained effect Effects 0.000 description 1
- 230000002123 temporal effect Effects 0.000 description 1
- 229930101283 tetracycline Natural products 0.000 description 1
- 229960002180 tetracycline Drugs 0.000 description 1
- 235000019364 tetracycline Nutrition 0.000 description 1
- 150000003522 tetracyclines Chemical class 0.000 description 1
- 208000008732 thymoma Diseases 0.000 description 1
- 229960005267 tositumomab Drugs 0.000 description 1
- 230000002103 transcriptional effect Effects 0.000 description 1
- 230000002463 transducing effect Effects 0.000 description 1
- 238000001890 transfection Methods 0.000 description 1
- 238000013519 translation Methods 0.000 description 1
- 238000002054 transplantation Methods 0.000 description 1
- 238000011277 treatment modality Methods 0.000 description 1
- 238000011269 treatment regimen Methods 0.000 description 1
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 description 1
- PZYFJWVGRGEWGO-UHFFFAOYSA-N trisodium;hydrogen peroxide;trioxido(oxo)vanadium Chemical compound [Na+].[Na+].[Na+].OO.OO.OO.[O-][V]([O-])([O-])=O PZYFJWVGRGEWGO-UHFFFAOYSA-N 0.000 description 1
- 230000001173 tumoral effect Effects 0.000 description 1
- OUYCCCASQSFEME-UHFFFAOYSA-N tyrosine Natural products OC(=O)C(N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-UHFFFAOYSA-N 0.000 description 1
- 108091005990 tyrosine-phosphorylated proteins Proteins 0.000 description 1
- 125000001493 tyrosinyl group Chemical group [H]OC1=C([H])C([H])=C(C([H])=C1[H])C([H])([H])C([H])(N([H])[H])C(*)=O 0.000 description 1
- 229950004593 ublituximab Drugs 0.000 description 1
- 230000003827 upregulation Effects 0.000 description 1
- 238000011144 upstream manufacturing Methods 0.000 description 1
- 230000009385 viral infection Effects 0.000 description 1
- 230000003442 weekly effect Effects 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/705—Receptors; Cell surface antigens; Cell surface determinants
- C07K14/70575—NGF/TNF-superfamily, e.g. CD70, CD95L, CD153, CD154
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
- A61K35/14—Blood; Artificial blood
- A61K35/17—Lymphocytes; B-cells; T-cells; Natural killer cells; Interferon-activated or cytokine-activated lymphocytes
-
- 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
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
- A61P35/02—Antineoplastic agents specific for leukemia
Definitions
- the present invention relates to methods and vaccine compositions for the treatment of B-cell malignancies.
- Epstein-Barr virus is a gamma-herpes virus that infects most of humans and has a marked tropism for B lymphocytes.
- EBV is known to be one of the strongest trigger of intrinsically uncontrolled B-cell proliferation and lymphomagenesis.
- Rare genetic diseases specifically predispose to defective control of EBV infection, leading to virus- associated hemophagocytic syndrome, lymphoproliferative disorders (LPD) such as Hodgkin and non-Hodgkin lymphomas 1 ' 2 .
- LPD lymphoproliferative disorders
- CTPS1, SH2D1A, MAGT1, ITK and CD27 have been associated with high penetrance of EBV infection with up to 90% of patients having developed diseases and lymphomas related to persistent EBV infection 3 ' 4 ' 5 ' 6 ' 7 ' 8 .
- Studies of these primary immunodeficiencies uncovered crucial pathways involved in T-cell response towards EBV-infected B lymphocytes and more generally in T-cell functions.
- efficiency of the immune response to EBV is indeed mainly dependent of the massive expansion of specific CD8 + cytotoxic T cells that eliminate EBV-infected B cells 9 ' 10 .
- XLP X-linked lymphoproliferative syndrome
- SLAM signaling lymphocytic activation molecule
- MAGT1 codes for a transmembrane Mg 2+ transporter involved in TCR signaling and expression of NKG2D, an important cytolytic activating cell receptor expressed by CD8 + T cells 14 ' 15 .
- NKG2D and SLAM-SAP pathways represent important components of the immune response to EBV.
- ITK deficiency is caused by mutations in ITK, a well-characterized tyrosine kinase of the BTK family involved in T-cell activation through its ability to activate the PLC- ⁇ 16 .
- PLC- ⁇ activation in response to TCR activation is compromised resulting in decreased Ca 2+ mobilization 17 ' 18 .
- Itk-deficient mice exhibit defective CD8 + T-cell expansion during anti- viral responses 19 .
- the exact mechanisms underlying the defective immune response to EBV in ITK-deficient patients is not known.
- CD27 A deficiency in CD27 has been recently identified in 17 patients who all developed EBV-driven lymphoproliferative disorders supportive of a key role of CD27 in immunity against EBV, although its exact mechanism of action has not been delineated so far 5 ' 6 ' 20 ' 21 ' 22 .
- the ligand of CD27, CD70 is expressed on some B lymphocytes and dendritic cells subpopulations, and transiently found on most immune cells when activated 23 .
- CD70 is also present on a variety of cancer cells including neoplasias of B-cell origin 24 .
- CD27 and CD70 are homodimer type I and homotrimer type II membrane proteins, respectively 21 . Both belong to the Tumor Necrosis Factor (TNF) superfamily.
- TNF Tumor Necrosis Factor
- the present invention relates to methods and vaccine compositions for the treatment of
- Epstein-Barr virus (EBV) infection in humans is a major trigger of malignant and non- malignant B cell proliferations.
- ITK and CD27 deficiencies are characterized by impaired immune responses to EBV-infected B cells, though the underlying pathological mechanisms have not yet been identified.
- the inventors report a patient suffering from recurrent EBV-induced B cell proliferation due to a deficiency in CD70, the ligand of CD27, a co-stimulatory molecule of T cells. They show that EBV-specific T lymphocytes cannot expand properly when stimulated with CD70-deficient EBV-infected B cells, while expression of CD70 restores expansion.
- CD70-CD27-ITK pathway appears to be a crucial component of EBV-specific T-cell immunity and more generally for the immune surveillance of B-cells. Accordingly, CD70-CD27 could represent an important target to induce anti-tumoral vaccination. Restoration or induction of CD70 expression in some lymphoma cells lacking CD70 might represent a therapeutic approach to induce a global and potent anti-tumoral immunity.
- the first object of the present invention relates to a method of treating a B- cell malignancy in a subject in need thereof comprising i) providing a sample of malignant B cells obtained from the subject ii) isolating and culturing a population of malignant B cells from the sample of step i), iii) introducing in the population malignant B cells of step ii) a nucleic acid molecule encoding for a CD70 polypeptide and iv) administering to the subject a therapeutically effective amount of the population of malignant B cells of step iii).
- B-cell malignancy includes any type of leukemia or lymphoma of B cells.
- B cell lymphoma refers to a cancer that arises in cells of the lymphatic system from B cells.
- B cells are white blood cells that develop from bone marrow and produce antibodies. They are also known as B lymphocytes.
- B-cell malignancies include, but are not limited to, non-Hodgkin's lymphoma, Burkitt's lymphoma, small lymphocytic lymphoma, primary effusion lymphoma, diffuse large B-cell lymphoma, splenic marginal zone lymphoma, MALT (mucosa-associated lymphoid tissue) lymphoma, hairy cell leukemia, chronic lymphocytic leukemia, B-cell prolymphocytic leukemia, B cell lymphomas (e.g. various forms of Hodgkin's disease, B cell non-Hodgkin's lymphoma (NHL) and related lymphomas (e.g.
- Waldenstrom's macroglobulinaemia also called lymphoplasmacytic lymphoma or immunocytoma or central nervous system lymphomas
- leukemias e.g. acute lymphoblastic leukemia (ALL), chronic lymphocytic leukemia (CLL; also termed B cell chronic lymphocytic leukemia BCLL), hairy cell leukemia and chronic myoblastic leukemia
- myelomas e.g. multiple myeloma.
- Additional B cell malignancies include small lymphocytic lymphoma, B cell prolymphocytic leukemia, lymphoplasmacytic lymphoma, splenic marginal zone lymphoma, plasma cell myeloma, solitary plasmacytoma of bone, extraosseous plasmacytoma, extra-nodal marginal zone B cell lymphoma of mucosa-associated (MALT) lymphoid tissue, nodal marginal zone B cell lymphoma, follicular lymphoma, mantle cell lymphoma, diffuse large B cell lymphoma, mediastinal (thymic) large B cell lymphoma, intravascular large B cell lymphoma, primary effusion lymphoma, Burkitt's lymphoma/leukemia, grey zone lymphoma, B cell proliferations of uncertain malignant potential, lymphomatoid granulomatosis, and post-transplant lymphoproliferative disorder.
- MALT mucosa-associated lymphoid tissue
- the B-cell lymphoma is caused by Epstein-Barr virus (EBV).
- EBV Epstein-Barr virus
- the B cell lymphoma is a diffuse large B-cell lymphoma associated with mutation in the gene encoding for CD70.
- CD70 is indeed one of the most frequently mutated genes in diffuse large B-cell lymphomas 42 ' 44 , 45 .
- treatment refers to both prophylactic or preventive treatment as well as curative or disease modifying treatment, including treatment of patient at risk of contracting the disease or suspected to have contracted the disease as well as patients who are ill or have been diagnosed as suffering from a disease or medical condition, and includes suppression of clinical relapse.
- the treatment may be administered to a subject having a medical disorder or who ultimately may acquire the disorder, in order to prevent, cure, delay the onset of, reduce the severity of, or ameliorate one or more symptoms of a disorder or recurring disorder, or in order to prolong the survival of a subject beyond that expected in the absence of such treatment.
- therapeutic regimen is meant the pattern of treatment of an illness, e.g., the pattern of dosing used during therapy.
- a therapeutic regimen may include an induction regimen and a maintenance regimen.
- the phrase “induction regimen” or “induction period” refers to a therapeutic regimen (or the portion of a therapeutic regimen) that is used for the initial treatment of a disease.
- the general goal of an induction regimen is to provide a high level of drug to a patient during the initial period of a treatment regimen.
- An induction regimen may employ (in part or in whole) a "loading regimen", which may include administering a greater dose of the drug than a physician would employ during a maintenance regimen, administering a drug more frequently than a physician would administer the drug during a maintenance regimen, or both.
- maintenance regimen refers to a therapeutic regimen (or the portion of a therapeutic regimen) that is used for the maintenance of a patient during treatment of an illness, e.g., to keep the patient in remission for long periods of time (months or years).
- a maintenance regimen may employ continuous therapy (e.g., administering a drug at a regular intervals, e.g., weekly, monthly, yearly, etc.) or intermittent therapy (e.g., interrupted treatment, intermittent treatment, treatment at relapse, or treatment upon achievement of a particular predetermined criteria [e.g., disease manifestation, etc.]).
- the sample of malignant B cells can be obtained from a number of sources, including peripheral blood mononuclear cells, bone marrow, lymph node tissue, cord blood, and tumoral tissue.
- the B cells can be obtained from a unit of blood collected from a subject using any number of techniques known to the skilled artisan, such as FICOLLTM (copolymers of sucrose and epichlorohydrin that may be used to prepare high density solutions) separation.
- FICOLLTM copolymers of sucrose and epichlorohydrin that may be used to prepare high density solutions
- cells from the circulating blood of an individual are obtained by apheresis or leukapheresis.
- the apheresis product typically contains lymphocytes, including T cells, monocytes, granulocytes, B cells, other nucleated white blood cells, red blood cells, and platelets.
- the cells collected by apheresis may be washed to remove the plasma fraction and to place the cells in an appropriate buffer or media for subsequent processing steps.
- the cells are washed with phosphate buffered saline (PBS).
- PBS phosphate buffered saline
- the wash solution lacks calcium and may lack magnesium or may lack many if not all divalent cations.
- a washing step may be accomplished by methods known to those in the art, such as by using a semi-automated "flow-through" centrifuge (for example, the Cobe 2991 cell processor) according to the manufacturer's instructions.
- B cells may be isolated from peripheral blood or leukapheresis using techniques known in the art.
- PBMCs may be isolated using FICOLLTM (Sigma-Aldrich, St Louis, Mo.) and CD 19+ B cells purified by negative or positive selection using any of a variety of antibodies known in the art, such as the Rosette tetrameric complex system (StemCell Technologies, Vancouver, Canada).
- FICOLLTM Sigma-Aldrich, St Louis, Mo.
- CD 19+ B cells purified by negative or positive selection using any of a variety of antibodies known in the art, such as the Rosette tetrameric complex system (StemCell Technologies, Vancouver, Canada).
- Other isolation kits are commercially available, such as R&D Systems' MagCellect Human B Cell Isolation Kit (Minneapolis, Minn.).
- CD70 has its general meaning in the art and refers to the ligand for CD27 (see, for example, Bowman M R et al., J. Immunol. 1994 Feb. 15; 152(4):1756- 61). CD70 is also referred to as “CD70 molecule”, “CD27L”, “CD27LG”, “TNFSF7,” “tumor necrosis factor (ligand) superfamily member 7," “CD27 ligand,” “CD70 antigen,” “surface antigen CD70,” “tumor necrosis factor ligand superfamily, member 7,” “Ki-24 antigen,” and "CD27-L”.
- CD70 is a type II transmembrane protein that belongs to the tumor necrosis factor (TNF) ligand family. It is a surface antigen on activated T and B lymphocytes that induces proliferation of co-stimulated T cells, enhances the generation of cytolytic T cells, and contributes to T cell activation. It has also been suggested that CD70 plays a role in regulating B-cell activation, cytotoxic function of natural killer cells, and immunoglobulin synthesis (Hintzen R Q et al., J. Immunol. 1994 Feb. 15; 152(4): 1762-73).
- An exemplary human amino acid sequence of CD70 is reference in GenBank under the Accession No. NP— 001243 (SEQ ID NO: 1):
- the malignant B cells are transformed with a nucleic acid molecule encoding for a polypeptide comprising an amino acid sequence having at least 90% of identity with SEQ ID NO: l
- a first amino acid sequence having at least 90% of identity with a second amino acid sequence means that the first sequence has 90; 91; 92; 93; 94; 95; 96; 97; 98; 99 or 100% of identity with the second amino acid sequence.
- Sequence identity is frequently measured in terms of percentage identity (or similarity or homology); the higher the percentage, the more similar are the two sequences.
- Methods of alignment of sequences for comparison are well known in the art. Various programs and alignment algorithms are described in: Smith and Waterman, Adv. Appl. Math., 2:482, 1981; Needleman and Wunsch, J. Mol. Biol, 48:443, 1970; Pearson and Lipman, Proc. Natl. Acad. Sci.
- the alignment tools ALIGN Myers and Miller, CABIOS 4: 11-17, 1989
- LFASTA Pearson and the University of Virginia, fasta20u63 version 2.0u63, release date December 1996
- ALIGN compares entire sequences against one another
- LFASTA compares regions of local similarity.
- these alignment tools and their respective tutorials are available on the Internet at the NCSA Website, for instance.
- the Blast 2 sequences function can be employed using the default BLOSUM62 matrix set to default parameters, (gap existence cost of 11, and a per residue gap cost of 1).
- the alignment should be performed using the Blast 2 sequences function, employing the PAM30 matrix set to default parameters (open gap 9, extension gap 1 penalties).
- the BLAST sequence comparison system is available, for instance, from the NCBI web site; see also Altschul et al, J. Mol. Biol, 215:403-410, 1990; Gish. & States, Nature Genet., 3:266-272, 1993; Madden et al. Meth. EnzymoL, 266: 131-141, 1996; Altschul et al, Nucleic Acids Res., 25:3389-3402, 1997; and Zhang & Madden, Genome Res., 7:649-656, 1997.
- nucleic acid molecule has its general meaning in the art and refers to a DNA or RNA molecule. However, the term captures sequences that include any of the known base analogues of DNA and RNA such as, but not limited to 4-acetylcytosine, 8- hydroxy-N6-methyladenosine, aziridinylcytosine, pseudoisocytosine, 5-
- transformation means the introduction of a "foreign” (i.e. extrinsic or extracellular) gene, DNA or RNA sequence to a host cell, so that the host cell will express the introduced gene or sequence to produce a desired substance, typically a protein or enzyme coded by the introduced gene or sequence.
- a host cell that receives and expresses introduced DNA or RNA has been "transformed”.
- the transformation of the malignant B cells with the nucleic acid molecule employs viral vectors to transduce B cells.
- viral vectors include, without limitation, adenovirus-based vectors, adeno-associated virus (AAV)-based vectors, retroviral vectors, retroviral-adenoviral vectors, and vectors derived from herpes simplex viruses (HSVs), including amplicon vectors, replication-defective HSV and attenuated HSV (see, e.g., Krisky, Gene Ther. 5: 1517-30, 1998; Pfeifer, Annu. Rev. Genomics Hum. Genet. 2: 177-211, 2001, each of which is incorporated by reference in its entirety).
- HSVs herpes simplex viruses
- the B cell are transduced with retroviral vectors, or vectors derived from retroviruses.
- retroviral vectors or vectors derived from retroviruses.
- retroviruses are enveloped RNA viruses that are capable of infecting animal cells, and that utilize the enzyme reverse transcriptase in the early stages of infection to generate a DNA copy from their RNA genome, which is then typically integrated into the host genome.
- retroviral vectors Moloney murine leukemia virus (MLV)-derived vectors, retroviral vectors based on a Murine Stem Cell Virus, which provides long-term stable expression in target cells such as hematopoietic precursor cells and their differentiated progeny (see, e.g., Hawley et al, PNAS USA 93: 10297-10302, 1996; Keller et al, Blood 92:877-887, 1998), hybrid vectors (see, e.g., Choi, et al, Stem Cells 19:236-246, 2001), and complex retrovirus-derived vectors, such as lentiviral vectors. As noted above, in some embodiments employ lentiviral vectors.
- MMV Moloney murine leukemia virus
- lentivirus refers to a genus of complex retroviruses that are capable of infecting both dividing and non-dividing cells.
- lentiviruses include HIV (human immunodeficiency virus; including HIV type 1, and HIV type 2), visna-maedi, the caprine arthritis-encephalitis virus, equine infectious anemia virus, feline immunodeficiency virus (FIV), bovine immune deficiency virus (BIV), and simian immunodeficiency virus (SIV).
- HIV human immunodeficiency virus
- HIV type 1 HIV type 2
- visna-maedi the caprine arthritis-encephalitis virus
- equine infectious anemia virus feline immunodeficiency virus (FIV), bovine immune deficiency virus (BIV), and simian immunodeficiency virus (SIV).
- Lentiviral vectors can be derived from any one or more of these lentiviruses (see, e.g., Evans e
- the retroviral vector comprises certain minimal sequences from a lentivirus genome, such as the HIV genome or the SIV genome.
- the genome of a lentivirus is typically organized into a 5 ' long terminal repeat (LTR) region, the gag gene, the pol gene, the env gene, the accessory genes (e.g., nef, vif, vpr, vpu, tat, rev) and a 3 ' LTR region.
- the viral LTR is divided into three regions referred to as U3, R (repeat) and U5.
- the U3 region contains the enhancer and promoter elements
- the U5 region contains the polyadenylation signals
- the R region separates the U3 and U5 regions.
- RNA Viruses A Practical Approach
- O Narayan J. Gen. Virology. 70: 1617-1639, 1989
- Fields et al Fundamental Virology Raven Press., 1990
- Miyoshi et al J Virol. 72:8150-7, 1998
- Lentiviral vectors may comprise any one or more of these elements of the lentiviral genome, to regulate the activity of the vector as desired, or, they may contain deletions, insertions, substitutions, or mutations in one or more of these elements, such as to reduce the pathological effects of lentiviral replication, or to limit the lentiviral vector to a single round of infection.
- a minimal retroviral vector comprises certain 5 f LTR and 3' LTR sequences, one or more genes of interest (to be expressed in the target cell), one or more promoters, and a cis-acting sequence for packaging of the RNA.
- Other regulatory sequences can be included, as described herein and known in the art.
- the viral vector is typically cloned into a plasmid that may be transfected into a packaging cell line, such as a eukaryotic cell (e.g., 293-HEK), and also typically comprises sequences useful for replication of the plasmid in bacteria.
- a packaging cell line such as a eukaryotic cell (e.g., 293-HEK)
- the nucleic acid molecule of t interest is located between the 5 f LTR and 3' LTR sequences.
- the nucleic acid molecule of interest is preferably in a functional relationship with other genetic elements, for example, transcription regulatory sequences such as promoters and/or enhancers, to regulate expression of the gene of interest in a particular manner once the gene is incorporated into the target cell.
- the useful transcriptional regulatory sequences are those that are highly regulated with respect to activity, both temporally and spatially.
- the viral vectors such as retroviral vectors employ one or more heterologous promoters, enhancers, or both.
- the U3 sequence from a retroviral or lentiviral 5 ' LTR may be replaced with a promoter or enhancer sequence in the viral construct.
- a “functional relationship” and “operably linked” mean, without limitation, that the gene is in the correct location and orientation with respect to the promoter and/or enhancer, such that expression of the gene will be affected when the promoter and/or enhancer is contacted with the appropriate regulatory molecules.
- Any enhancer/promoter combination may be used that either regulates (e.g., increases, decreases) expression of the viral RNA genome in the packaging cell line, regulates expression of the selected gene of interest in an infected target cell, or both.
- a promoter is an expression control element formed by a DNA sequence that permits binding of RNA polymerase and transcription to occur.
- Promoters are untranslated sequences that are located upstream (5 ' ) of the start codon of a selected gene of interest (typically within about 100 to 1000 bp) and control the transcription and translation of the coding polynucleotide sequence to which they are operably linked. Promoters may be inducible or constitutive. Inducible promoters initiate increased levels of transcription from DNA under their control in response to some change in culture conditions, such as a change in temperature. A variety of promoters are known in the art, as are methods for operably linking the promoter to the polynucleotide coding sequence. Both native promoter sequences and many heterologous promoters may be used to direct expression of the selected gene of interest.
- the promoter is a heterologous promoters, because they generally permit greater transcription and higher yields of the desired protein as compared to the native promoter.
- the promoter is selected among heterologous viral promoters. Examples of such promoters include those obtained from the genomes of viruses such as polyoma virus, fowlpox virus, adenovirus, bovine papilloma virus, avian sarcoma virus, cytomegalovirus, a retrovirus, hepatitis-B virus and Simian Virus 40 (SV40).
- viruses such as polyoma virus, fowlpox virus, adenovirus, bovine papilloma virus, avian sarcoma virus, cytomegalovirus, a retrovirus, hepatitis-B virus and Simian Virus 40 (SV40).
- the promoter is a heterologous mammalian promoter, such as the actin promoter, an immunoglobulin promoter, a heat-shock promoter, or a promoter that is associated with the native sequence of the gene of interest.
- the promoter is compatible with the target cell, such as a quiescent B-lymphocyte, an activated B-lymphocyte, a plasma B cell, or a memory B cell.
- constitutive promoters include the promoter for ubiquitin, the CMV promoter (see, e.g., Karasuyama et al, J. Exp. Med.
- tissue specific promoters include the Ick promoter (see, e.g., Garvin et al, Mol. Cell Biol. 8:3058-3064, 1988; and Takadera et al, Mol. Cell Biol.
- promoters include the ubiquitin-C promoter, the human ⁇ heavy chain promoter or the Ig heavy chain promoter (e.g., MH-M2), and the human ⁇ light chain promoter or the Ig light chain promoter (e.g., EEK-M2), which are functional in B-lymphocytes.
- promoters may be selected to allow for inducible expression of the gene.
- a number of systems for inducible expression are known in the art, including the tetracycline responsive system and the lac operator-repressor system. It is also contemplated that a combination of promoters may be used to obtain the desired expression of the gene of interest. The skilled artisan will be able to select a promoter based on the desired expression pattern of the gene in the organism and/or the target cell of interest.
- the viral vectors e.g., retroviral, lentiviral
- the viral vectors are "pseudo-typed" with one or more selected viral glycoproteins or envelope proteins, mainly to target selected cell types.
- Pseudo-typing refers to generally to the incorporation of one or more heterologous viral glycoproteins onto the cell-surface virus particle, often allowing the virus particle to infect a selected cell that differs from its normal target cells.
- a “heterologous” element is derived from a virus other than the virus from which the RNA genome of the viral vector is derived.
- the glycoprotein-coding regions of the viral vector have been genetically altered such as by deletion to prevent expression of its own glycoprotein.
- the envelope glycoproteins gp41 and/or gpl20 from an HIV-derived lentiviral vector are typically deleted prior to pseudo-typing with a heterologous viral glycoprotein.
- the viral vector is pseudo-typed with a heterologous viral glycoprotein that targets plasma cells such as B-lymphocytes.
- the viral glycoprotein allows selective infection or transduction of resting or quiescent B-lymphocytes.
- the viral glycoprotein allows selective infection of activated B- lymphocytes.
- the viral glycoprotein allows infection or transduction of both quiescent B-lymphocytes and activated B-lymphocytes.
- viral glycoprotein allows infection of B cell chronic lymphocyte leukemia cells.
- the heterologous viral glycoprotein is derived from the glycoprotein of the measles virus, such as the Edmonton measles virus.
- the viral vector comprises an embedded antibody binding domain, such as one or more variable regions (e.g., heavy and light chain variable regions) which serves to target the vector to a particular cell type.
- Generation of viral vectors can be accomplished using any suitable genetic engineering techniques known in the art, including, without limitation, the standard techniques of restriction endonuclease digestion, ligation, transformation, plasmid purification, PCR amplification, and DNA sequencing, for example as described in Sambrook et al. (Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory Press, N.Y. (1989)), Coffin et al. (Retroviruses. Cold Spring Harbor Laboratory Press, N.Y. (1997)) and "RNA Viruses: A Practical Approach” (Alan J. Cann, Ed., Oxford University Press, (2000)).
- the B cells may be transduced with the viral vectors described herein using any of a variety of known techniques in the art (see, e.g., Science 12 Apr. 1996 272: 263-267; Blood 2007, 99:2342-2350; Blood 2009, 113: 1422-1431; Blood 2009 Oct. 8; 114(15):3173-80; Blood. 2003; 101(6) :2167-2174; Current Protocols in Molecular Biology or Current Protocols in Immunology, John Wiley & Sons, New York, N.Y. (2009)).
- PBMCs, B- or T- lymphocytes from donors and other B cell cancer cells such as B-CLLs may be isolated and cultured in RPMI 1640 (GibcoBRL Invitrogen, Auckland, New Zealand) or other suitable medium, either serum- free or supplemented with 10% FCS and penicillin/streptomycin and/or other suitable supplements.
- RPMI 1640 GibcoBRL Invitrogen, Auckland, New Zealand
- FCS fetal bovine serum
- penicillin/streptomycin and/or other suitable supplements fetal bovine serum
- cells are seeded at 10 5 cells in 48-wellplates and concentrated vector added at various doses that may be routinely optimized by the skilled person using routine methodologies.
- B cells may be transferred to MS5 cell monolayer in RPMI supplemented with 10% AB serum, 5% FCS, 50 ng/ml rhSCF, 10 ng/ml rhIL-15 and 5 ng/ml rhIL-2 and medium refreshed periodically as needed.
- suitable media and supplements may be used as desired.
- the B cells are contacted with a retroviral vector as described herein comprising a nucleic acid of interest operably linked to a promoter, under conditions sufficient to transduce at least a portion of the B cells. In some embodiments the B cells are contacted with a retroviral vector as described herein comprising a nucleic acid of interest operably linked to a promoter, under conditions sufficient to transduce at least 20% of the B cells.
- the B cells are contacted with a vector as described herein under conditions sufficient to transduce at least 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or even 100% of the B cells.
- Any of a variety of culture media may be used in the present methods as would be known to the skilled person (see e.g., Current Protocols in Cell Culture, 2000-2009 by John Wiley & Sons, Inc.).
- media for use in the methods described herein includes, but is not limited to Iscove modified Dulbecco medium (with or without fetal bovine or other appropriate serum).
- Illustrative media also includes, but is not limited to, RPMI 1640, AIM-V, DMEM, MEM, a-MEM, F-12, X-Vivo 15, and X-Vivo 20.
- the medium may comprise a surfactant, an antibody, plasmanate or a reducing agent (e.g. N-acetyl-cysteine, 2-mercaptoethanol), or one or more antibiotics.
- a reducing agent e.g. N-acetyl-cysteine, 2-mercaptoethanol
- IL-6, soluble CD40L, and a cross-linking enhancer may also used.
- cells are cultured for 1-7 days. In some embodiments, cells are cultured 7, 14, 21 days or longer. Thus, cells may be cultured under appropriate conditions for 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or more days. Cells are replated, media and supplements may be added or changed as needed using techniques known in the art.
- the transduced B cells may be cultured under conditions and for sufficient time periods such that at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%), 98%o, 99%) or 100% of the cells express the transgene of interest.
- the population of malignant B cells transformed with the nucleic acid molecule encoding for the CD70 polypeptide is thus particularly suitable for inducting anti-tumoral vaccination.
- said population of cells will promote the T-cell mediated immunity against tumor cells.
- the population of malignant cells transformed according to the invention may be administered either alone, or as a vaccine composition in combination with diluents and/or with other components such as cytokines or cell populations.
- the vaccine compositions of the present invention the population of transformed malignant B cells in combination with one or more pharmaceutically or physiologically acceptable carriers, diluents or excipients.
- compositions may comprise buffers such as neutral buffered saline, phosphate buffered saline and the like; carbohydrates such as glucose, mannose, sucrose or dextrans, mannitol; proteins; polypeptides or amino acids such as glycine; antioxidants; chelating agents such as EDTA or glutathione; adjuvants (e.g., aluminum hydroxide); and preservatives.
- buffers such as neutral buffered saline, phosphate buffered saline and the like
- carbohydrates such as glucose, mannose, sucrose or dextrans, mannitol
- proteins polypeptides or amino acids such as glycine
- antioxidants e.g., chelating agents such as EDTA or glutathione
- adjuvants e.g., aluminum hydroxide
- preservatives e.g., aluminum hydroxide
- a therapeutically effective amount an anti-tumor effective amount
- a tumor-inhibiting effective amount or “therapeutic amount”
- the precise amount of the compositions of the present invention to be administered can be determined by a physician with consideration of individual differences in age, weight, tumor size, extent of infection or metastasis, and condition of the patient (subject). It can generally be stated that a vaccine composition comprising the B cells described herein may be administered at a dosage of 10 4 to 10 7 cells/kg body weight, preferably 10 5 to 10 6 cells/kg body weight, including all integer values within those ranges.
- Vaccine compositions may also be administered multiple times at these dosages.
- the cells can be administered by using infusion techniques that are commonly known in immunotherapy (see, e.g., Rosenberg et al, New Eng. J. of Med. 319:1676, 1988).
- the optimal dosage and treatment regime for a particular patient can readily be determined by one skilled in the art of medicine by monitoring the patient for signs of disease and adjusting the treatment accordingly.
- antigen-specific T cells are administered approximately at 2> ⁇ 10 9 to 2x lO n cells to the patient. (See, e.g., U.S. Pat. No. 5,057,423).
- lower numbers of the transduced B cells of the present invention in the range of 10 6 /kilogram (10 6 -10 u per patient) may be administered.
- the B cells are administered at 10 5 , 10 6 , 10 7 , 10 8 , 10 9 , 10 10 , 10 11 , or 10 12 cells to the subject.
- the vaccine compositions may be administered multiple times at dosages within these ranges.
- the administration of the subject of the vaccine compositions may be carried out in any convenient manner, including by aerosol inhalation, injection, ingestion, transfusion, implantation or transplantation.
- compositions described herein may be administered to a patient subcutaneously, intradermally, intratumorally, intranodally, intramedullary, intramuscularly, by intravenous (i.v.) injection, or intraperitoneally.
- the vaccine compositions of the present invention are administered to a patient by intradermal or subcutaneous injection.
- the vaccine compositions as described herein are preferably administered by i.v. injection.
- the vaccine composition may be injected directly into a tumor, or lymph node.
- the vaccine composition is administered to the subject in conjunction with (e.g. before, simultaneously or following) any number of relevant treatment modalities, including but not limited to treatment with agents such as antiviral agents, chemotherapy, radiation, immunosuppressive agents, such as cyclosporin, azathioprine, methotrexate, mycophenolate, and FK506, antibodies, or other immunoablative agents such as CAMPATH, anti-CD3 antibodies or other antibody therapies, cytoxin, fludaribine, cyclosporin, FK506, rapamycin, mycophenolic acid, steroids, FR901228, cytokines, and irradiation.
- agents such as antiviral agents, chemotherapy, radiation, immunosuppressive agents, such as cyclosporin, azathioprine, methotrexate, mycophenolate, and FK506, antibodies, or other immunoablative agents such as CAMPATH, anti-CD3 antibodies or other antibody therapies, cytoxin, fludaribine, cyclosporin, F
- the vaccine compositions of the present invention are administered to a patient in conjunction with (e.g.
- the vaccine compositions of the present invention are administered following B- cell ablative therapy such as agents that react with CD20.
- B- cell ablative therapy such as agents that react with CD20.
- antibodies having specificity for CD20 include: “C2B8” which is now called “Rituximab” ("RITUXAN®”) (U.S. Pat. No.
- murine IgGl kappa mAb covalently linked to MX-DTPA for chelating to yttrium-[90] murine IgG2a "BI,” also called “Tositumomab,” optionally labeled with radioactive 1311 to generate the "1311-B1" antibody (iodine 131 tositumomab, BEXXARTM) (U.S. Pat. No. 5,595,721, expressly incorporated herein by reference); murine monoclonal antibody "1F5" (Press et al.
- AME-133 (ocaratuzumab; Applied Molecular Evolution), a a fully- humanized and optimized IgGl mAb against CD20; A20 antibody or variants thereof such as chimeric or humanized A20 antibody (cA20, bA20, respectively) (U.S. Ser. No. 10/366,709, expressly incorporated herein by reference, Immunomedics); and monoclonal antibodies L27, G28-2, 93-1B3, B-CI or NU-B2 available from the International Leukocyte Typing Workshop (Valentine et al, In: Leukocyte Typing III (McMichael, Ed., p.
- suitable antibodies include e.g. antibody GAlOl (obinutuzumab), a third generation humanized anti-CD20-antibody of Biogen Idec/Genentech/Roche.
- BLX- 301 of Biolex Therapeutics a humanized anti CD20 with optimized glycosylation or Veltuzumab (bA20), a 2nd-generation humanized antibody specific for CD20 of Immunomedics or DXL625, derivatives of veltuzumab, such as the bispecific hexavalent antibodies of IBC Pharmaceuticals (Immunomedics) which are comprised of a divalent anti- CD20 IgG of veltuzumab and a pair of stabilized dimers of Fab derived from milatuzumab, an anti-CD20 mAb enhanced with InNexus' Dynamic Cross Linking technology, of Inexus Biotechnology both are humanized anti-CD20 antibodies are suitable.
- BM-ca a humanized antibody specific for CD20 (Int J. Oncol. 2011 February; 38(2):335-44)), C2H7 (a chimeric antibody specific for CD20 (Mol Immunol. 2008 May; 45(10):2861-8)), PR0131921 (a third generation antibody specific for CD20 developed by Genentech), Reditux (a biosimilar version of rituximab developed by Dr Reddy's), PBO-326 (a biosimilar version of rituximab developed by Probiomed), a biosimilar version of rituximab developed by Zenotech, TL-011 (a biosimilar version of rituximab developed by Teva), CMAB304 (a biosimilar version of rituximab developed by Shanghai CP Guojian), GP-2013 (a biosimilar version of rituximab developed by Sandoz (Novartis)), SAIT-101 (a biosimilar version of rituximab developed by Samsung BioLogic
- FIGURES are a diagrammatic representation of FIGURES.
- T cells (a) Cytotoxic response of T cells from two control individuals (Ctr. l and Ctr.2) and the CD70-deficient patient (Pat.) against autologous LCLs as measured by Cr 51 release at the indicated effector-to-target (E:T) ratios. T cells have been co cultured with the autologous LCLs for 4 weeks before the test. One representative of two independent experiments. Triplicates with s.d.
- CD70 (pLenti vector). One representative of five independent experiments.
- Figure 2 Analysis of cytolytic activity of T cells expanded from PBMCs of one healthy control (Ctr.) and the CD70-deficient patient (Pat.) for 8-15 days with irradiated autologous CD70-expressing (empty CRISPR or pLenti-CD70) or CD70-deficient (CD70- CRISPR or empty vector) LCL cells. Cytolytic activity of T cells was then tested against a mixture of autologous CD70-expressing (in blue) or CD70-deficient (in red) LCL cells as target cells at a ratio effector-to-target 1 : 1 of 1 for 0, 4 and 12 hours. Residual target cells were evaluated by FACS analysis. Data are represented in percentages of cells normalized to the percentages at time 0.
- the two patients carriers of the homozygous c.85C>T, p.R29C in ITK are issued from consanguineous parents of Amsterdamn origin. ITK gene defects were identified by WES and identified mutations were further verified by direct DNA sequencing.
- the patient carrier of the homozygous c.329G>A, p.Wl 10X in CD27 is issued from consanguineous parents of Tunisian origin and suffered from a recurrent EBV-driven lymphoproliferative disease.
- the CD27 defect was diagnosed based on the lack of CD27 expression on B cells. The four exons of CD27 were further sequenced.
- Exome sequencing and analysis Exome sequencing and analysis. Exome capture was performed according to the manufacturer's protocol using the Illumina TruSeq exome enrichment kit and sequencing of 100 bp paired end reads on an Illumina HiSeq. Approximately 10 Gb of sequence were obtained for each subject such that 90% of the coding bases of the exome defined by the consensus coding sequence (CCDS) project were covered by at least 10 reads. Adaptor sequences and quality trimmed reads were removed using the Fastx toolkit (http ://hannonlab . cshl.edu/fastx_toolkit/) and a custom script was then used to ensure that only read pairs with both mates present were subsequently used.
- CCDS consensus coding sequence
- Reads were aligned to hgl9 with BWA31, and duplicate reads were marked using Pi card (http://picard.sourceforge.net/) and excluded from downstream analyses.
- Single nucleotide variants (SNVs) and short insertions and deletions (indels) were determined using samtools (http://samtools.sourceforge.net/) pileup and varFilter32 with the base alignment quality (BAQ) adjustment disabled, they were then quality filtered to require at least 20% of reads supporting the variant call.
- Variants were annotated using both A NOVAR33 and custom scripts to identify whether they affected protein coding sequences, and whether they had previously been seen in the public data bases of exomes and the 7566 exomes previously sequenced at our center.
- Genomic DNA from peripheral blood cells of the patient, their parents, and other family members was isolated according to standard methods. Oligonucleotide primers in the introns flanking the exon 3 of CD70, the exon 1 oflTK and the exon 3 of CD27 were used to amplify genomic DNA.
- PCR products were amplified using high fidelity Platinum Taq DNA Polymerase (Invitrogen) according to the manufacturer's recommendations, purified with the QIAquick gel extraction kit (Qiagen), sequenced using the ABI PRISM BigDye Terminator Cycle Sequencing Ready Reaction Kit (PerkinElmer) according to the manufacturer's recommendations and analyzed with 3500xL Genetic Analyzer (Applied Biosystems). All collected sequences were analyzed using 4peaks software (Version 1.7.2; A. Griekspoor and T. Groothuis, http://mekentosj.com/4peaks/) or DNADynamo (BlueTractorSoftware).
- PBMCs Peripheral blood mononuclear cells
- PHA phytohaemagglutinin
- Panserin 401 Pan Biotech
- BioWest penicillin
- penicillin 100 U ml "1
- streptomycin 100 ⁇ g ml "1 ).
- the Burkitt's lymphoma cell line Raji and the mouse thymoma L1210 obtained from ATCC were cultured in RPMI 1640 GlutaMax medium (Invitrogen) supplemented with 10% heat-inactivated fetal calf serum (Gibco), penicillin (100U ml "1 ) and streptomycin (100 ⁇ ml "1 ). Cells were free of mycoplasma and tested for mycoplasma contamination on a regular basis.
- EBV-transformed LCLs and EBV-specific T cell lines were generated from fresh or frozen PBMCs of the patient and control donors. The PBMCs were incubated with supernatant from B95-8 cells in the presence of ⁇ g/ml of cyclosporine A (Sigma- Aldrich) as previously described 49 . EBV-specific T cell lines were generated from the patient and control healthy donors. PBMCs were co- cultured with 40 Gy irradiated autologous LCLs at an effector-to-stimulator (E/S) ratio of 40: 1. After 9-14 days, viable cells were stimulated with 40 Gy irradiated autologous LCLs at E/S ratio 4: 1 for 5-7days. And then analyzed for cytotoxicity.
- E/S effector-to-stimulator
- Cytolytic activity of T cells expanded for 8 days from PBMCs was also evaluated in co cultures against autologous cell trace violet-labeled CD70-expressing LCLs and CFSE-labeled CD70-deficient LCLs, as at a ratio effector-to-target of 1 :1 for 0, 4 and 12 hours. Residual target cells were evaluated by FACS analysis. Data are represented in percentages of cells normalized to the percentages at time 0. Degranulation of CD8 + T cells was determined by analysis of the expression of CD107/LAMP, a marker of the exocytosis of lytic granules as described before 8 .
- PHA-stimulated T cells were washed and cultured without IL-2 for 72 hours to synchronize the cells. Then PHA-stimulated T cells or PBMCs were cultured during 4 to 8 days in complete medium alone or in the presence of anti- CD3/CD28-coated beads (Invitrogen).
- PHA-stimulated T cells or PBMCs plus or minus anti-CD27 antibody (clone LG.3A10 from eBioscience or 0323 from BioLegend) (K ⁇ g.ml -1 ) from or ibrutinib (5 to 500nM) (Selleckchem), were co- cultured in the presence of 45 Gy irradiated autologous or allogenous LCLs that have been previously incubated with ⁇ g ml "1 of anti-CD3 antibody (OKT3) or not for one hour. Cells were co-cultured at ratio of one T cells or PBMC for 10 LCLs (ratio 1/10).
- Cell-proliferation was monitored by labeling T cells or PBMCs with the CellTrace violet dye (Violet Proliferation Dye 450, BD Biosciences) prior to stimulation or co-culture with LCLs, according to the manufacturer's instructions. After 8 days of culture, cells were harvested and CellTrace violet dye dilution was assessed by flow cytometry.
- CellTrace violet dye Violet Proliferation Dye 450, BD Biosciences
- anti-CD 16 (3G8), anti-CD45RA (HI 100), anti-CD45PvO (UCHL1), anti-CD95 (DX2), anti-TCRalphabeta, IgD (IA6-2), IgM (G20-127) from BD biosciences and anti-CD21 (BL13), anti-TCRgammadelta), anti-TCR Valpha24 (CI 5), anti-TCR Vbeta from Beckman coulter. Binding of CD27 to CD70 expressing-cells was assessed by incubation with Human CD27-muIg/Biotin Fusion Protein (Ancell Corporation) according to manufacturer's protocol and standard flow cytometry methods.
- Cytokine production and detection For intracellular staining of cytokines, cells were re-stimulated overnight with irradiated LCLs beforehand preincubated with ⁇ g ml "1 of OKT3 and in the presence brefeldin A (GolgiPlug, BD). Cells are then fixed and permeabilized using the BD cytofix/cytoperm plus kit (BD Pharmingen) according to manufacturer's instructions.
- BD Pharmingen BD cytofix/cytoperm plus kit
- cells are labeled with PE/Cy7-anti-TNF-a (mouse IgGl; MAbl l), APC-anti-IFN- ⁇ (mouse IgGl, 4S.B3) and isotype-matched monoclonal antibodies purchased from Bio-Legend and analyzed by flow cytometry. Thereafter, cells were collected, washed and stained with BV785-anti-CD3, BV650-PE-anti-CD8 and BV510-anti-CD4 mAbs and analyzed by flow cytometry.
- EBV-specific T cells detection EBV-specific CD8 + T cells from PBMCs were co- cultured with 45 Gy irradiated LCLs for 8-10 days and were detected using a mix of unlabelled EBV HLA-A2:01 Pro5® Pentamers (Proimmune) mixed with R-PE Pro5® Fluorotag in addition with BV785-anti-CD3, BV650-PE-anti-CD8 and BV510-anti-CD4 monoclonal antibodies.
- the EBV HLA-A2:01 Pro5® Pentamers mix contains 3 different pentamers presenting FLYALLALLL (residues 356-364 from LMP2), CLGGLLTMV (residues 426-434 from LMP2) or GLCTLVAML (residues 259-267 from BMLF-1) peptides derived from LMP2 and BMLF1 proteins of EBV.
- FLYALLALLL residues 356-364 from LMP2
- CLGGLLTMV residues 426-434 from LMP2
- GLCTLVAML residues 259-267 from BMLF-1 peptides derived from LMP2 and BMLF1 proteins of EBV.
- EBV-specific T cells activated phenotype is characterized using PE/Cy7-anti-CD25 and FITC-CD45RA antibodies. All staining are done according to manufacturer's instructions.
- the lentiCRISPR plasmid was a gift from Feng Zhang (Addgene plasmid # 49535). All sgRNAs were designed using M IT CRISPR Design Tool (http ://crispr .mit. edu) . Three pairs of 24-bp forward (F) and reverse (R) of oligonucleotides targeting different sequences in the exon 3 of CD70 were synthesized (Eurogentec) with a 4-bp overhang to enable cloning into the Bsmbl site in reverse oligonucleotides and a 4-bp overhang containing the PAM sequence in forward oligonucleotides. sgRNAs sequences.
- Pairs of synthetized oligos was annealed, phosphorylated, ligated to linearized vector and transformed into Stbl3 bacteria (Life Technologies). sgRNAs insertion was confirmed by Sanger sequencing using sequencing primer.
- the lentiCRISPR plasmids were transduced by infection in LCLs and transfected by electroporation with Nepa21 electroporator (Nepagene) in Raji cells.
- LCLs CD70-positive and CD70-negative populations were enriched by sorting.
- Protein concentrations were quantitated by BCA assay (BIO-RAD). 80 ⁇ g of proteins were separated by SDS-PAGE and transferred on PVDF membranes (Millipore). For testing expressiMembranes were blocked with milk or BSA for 1 h before incubation with primary antibodies for 90 minutes.
- anti-ITK (clone 2F12), anti-phosphorylated PLC- ⁇ (clone D6M9S), anti-phosphorylated ER l/2 (clone D13.14.4E) and anti-phosphorylated tyrosine (clone PY- 100) purchased from Cell signaling and, anti-FLAG (clone M2) and rabbit polyclonal anti- ACTIN antibody was from Sigma.
- Membranes were then washed and incubated with anti- mouse or anti-rabbit HRP-conjugated secondary antibodies from Cell Signaling and GE Healthcare, respectively. Pierce ECL western blotting substrate was used for detection.
- Binding assays using GST proteins were performed as outlined previously 50 , using lysates from unstimulated cells, anti-CD3 or pervanadate stimulated cells.
- GST-fusion proteins with the SH3 domains of human FYN, ITK and LCK were obtained by RT-PCR and subcloning into pGEX- 2T vector (GE Healthcare).
- CD70 is known to be the ligand for CD27 molecule. Deficiency in CD27 causes a high susceptibility to EBV infection and the associated lymphoproliferative disorders 5 ' 6 ' 20 . Thus, we considered CD70 as a strong candidate gene underlying the immunodeficiency of the patient.
- the c.535C>T mutation had no impact on the amount of CD70 mR A detected by qPCR in the PHA-stimulated T cells and in EBV-transformed B cells from the patient (further designated as lymphoblastoid cell line (LCL)) (data not shown).
- anti-CD70 antibody failed to detect CD70 by flow cytometry at cell surface of PHA-stimulated T cells of the patient.
- expression of CD70 was detectable on a fraction of PHA-stimulated T cells from healthy donors at day 8. This proportion of CD70 + T cells increased in culture with a large proportion of T cells expressing CD70 at day 15.
- CD70 was not detected on LCLs derived from the patient, in contrast to LCLs from healthy donors that expressed high levels of CD70 on their surface.
- Defective expression of CD70 was confirmed by analyzing the capacity of CD70 R179X to bind to CD27.
- a fusion protein containing the extracellular domain of CD27 (Fc-CD27) failed to bind on the surface of PHA-T cell blasts and LCLs from the patient, whereas its binding on control cells was detected. Similar results were obtained when wild-type CD70 or CD70 R179X proteins were transiently expressed in HEK-293 T cells. Expression of CD70 and CD70 R179X in HEK-293 T cells was further examined by western blot using N-terminus FLAG-tagged CD70 forms.
- the mutant CD70 R179X was weakly expressed compared to wild type CD70. Taken together, these results indicate that the p.R179X mutation in CD70 compromises its expression and has a deleterious effect on its ability to recognize its cognate ligand CD27. Therefore, we conclude that the CD70 deficiency mimics that of CD27 deficiency and thus likely accounts for the high susceptibility to EBV infection of the patient.
- CD70 expression has been shown to be restricted to some DCs and B cell subsets, while CD27 is expressed on most T cells and memory B cells 23 ' 29 . Indeed, expression of CD70 was barely detectable on CD4 and CD8 T cells, monocytes, DC cells and neutrophils, with the noticeable exception of a small fraction of B cells. This contrasts with the high levels of CD27 on the surface of CD4 + and CD8 + T lymphocytes and a small fraction of B cells.
- CD70 expression on B cells was rapidly upregulated upon activation by a combination of phorbol 12- myristate 13-acetate (PMA) and ionomycin (Iono). At day 3 of stimulation, more than 80% of B cells expressed large amounts of CD70 in contrast to T cells. Similarly to activated B cells, all EBV-transformed B cell lines that we tested expressed high levels of CD70. These data suggest that the expression of CD70 is inducible in B cells in the course of EBV infection. To test this possibility, we analyzed the expression of CD70 in tonsils from two individuals with infectious mononucleosis.
- CD70 staining was found on large cells that were positive for Epstein-Barr Encoded RNA (EBER) and PAX5 specific markers of EBV and B cells respectively and corresponded to EBV -infected B cells.
- EBER Epstein-Barr Encoded RNA
- PAX5 PAX5 specific markers of EBV and B cells respectively and corresponded to EBV -infected B cells.
- CD27 expression was not associated with PAX5 and EBER staining and accumulated in cells located in the T-cell areas.
- mice CD70 and CD27 play an important role in antigen specific-T cell responses, in particular during anti-viral responses 26 ' 30 .
- T-cell responses against EBV might be impaired in the absence of CD70.
- T cells of the patient Compared to control T-cell cultures, T cells of the patient exhibited a markedly decreased cytotoxic activity and IFN- ⁇ production, indicative of impaired T-cell responses to EBV.
- cytotoxic activity and IFN- ⁇ production indicative of impaired T-cell responses to EBV.
- reconstitution experiments by transducing LCLs of the patient with a lentiviral vector containing a cDNA coding wild-type CD70 (or an empty vector), which induced surface expression of CD70 to levels comparable to those seen on LCLs from control donors (Fig. lb).
- CD70-deficient LCLs were derived from PBMCs of a healthy HLA- A* 02 individual by gene inactivation. Most HLA- A* 02 individuals develop EBV-specific T cells against the GLCTLVAMV peptide (also termed GLC epitope), which is derived from the EBV-lytic cycle protein BMLF1 31 .
- GLCTLVAMV peptide also termed GLC epitope
- CD70-deficient LCLs were obtained by blunting CD70 expression using CRISPR (clustered regularly interspaced short palindrome repeats)-associated nuclease Cas9 technology.
- CD70KO- CRISPR1, CD70KO-CRISPR2 and CD70KO-CRISPR3 Three different CRISPR-Cas9 constructs containing RNA guides targeting exon 3 of CD70 were transiently transfected in the LCLs. After having been selected and sorted to enrich for CD70-negative cells, three stable CD70-deficient (CD70KO) LCLs cell lines were obtained (hereafter referred as CD70KO- CRISPR1, CD70KO-CRISPR2 and CD70KO-CRISPR3). These cell lines expressed comparable amounts of HLA- A* 02 molecules on their surface when compared to those of the wild-type parental LCLs or LCLs transfected with an empty CRISPR-Cas9 construct.
- EBV-specific T cells of the HLA-A*02 healthy donor were detected in coculture experiments with autologous irradiated wild-type LCLs (transfected with an empty CRISPR- Cas9 construct) or CD70KO-CRISPR1 LCLs.
- EBV-specific T cells were detectable at day 0 using a mix of pentamers for GLC, CLG and FLY epitopes, and following co culture with wild- type CD70-expressing LCLs (Empty CRISPR LCLs), their proportion increased at day 8 of culture by 5-fold. Most of these cells expressed CD25 and were strongly proliferative (data not shown).
- control and patient T cells expanded in the presence of autologous CD70-expressing LCLs were able to kill efficiently both CD70-expressing and CD70-deficient LCLs, indicating that CD70 deficiency per se does not preclude cytotoxic activity of T cells (Figure 2).
- T cells that were co cultured with autologous CD70-deficient LCLs had no killing activity demonstrating that specific effector T-cells require CD70 for expansion.
- CD70 on B cells provides a costimulatory signal required for TCR-mediated proliferation
- CD27-mediated T cell proliferation is dependent of ITK Because CD27, ITK, and CD70 deficiencies appear as phenocopies, we hypothesized that CD27-CD70 and ITK could form a functional and molecular cluster. To test it, we first examine CD70-dependent T-cell proliferation of PBMCs from CD27-deficient and ITK- deficient patients: four newly described patients, including one patient carrier of a homozygous mutation p.Wl 10X in CD27, two patients with a homozygous mutation p.R29C in ITK (Pat.l ITK " and Pat.2 ITK " ) and one patient with a homozygous mutation p.L38P in ITK (Pat.3 ITK " ).
- the Wl 10X mutation abolished the surface expression of CD27 on PBMCs and T-cell blasts of the patient.
- proliferation of CD27-deficient T-cell blasts in response to irradiated CD70-expressing B-LCLs in the presence of anti-CD3 antibody was markedly reduced.
- Expression of ITK was strongly diminished in lysates from T cells of patients carrying R29C and the L38P mutation.
- T-cell proliferation of ITK-deficient PBMCs or T-cell blasts in response to irradiated CD70-expressing LCLs in the presence of anti-CD3 antibody was markedly reduced.
- ITK-deficient T cells expressed CD27 levels comparable to those of control T cells or CD70-deficient T cells and displayed normal proliferation in response to CD3 or CD3 plus CD28 stimulations.
- ITK costimulatory signal triggered by CD27 on T cells when engaged by CD70 on B cells is dependent of the tyrosine kinase ITK.
- Ibrutinib an inhibitor of TEC kinases 32 including ITK inhibited CD70-expressing LCL-mediated T-cell proliferation.
- ITK is also known to activate PLC- ⁇ and the subsequent calcium flux following TCR activation 16 ' 18 .
- Calcium mobilization in T cells in response to stimulation by anti-CD3 antibody or anti-CD27 antibody was thus investigated.
- ITK-deficient PBMC derived T cells and T-cell blasts exhibited reduced calcium mobilization upon CD3 stimulation, in contrast to cells from healthy donors, CD70-deficient or CD27-deficient patients.
- CD27 signaling has not been yet clearly defined although a role for TRAF molecules has been reported 35 .
- CD27 contained in its intracellular domain several proline and tyrosine residues, which may form potential docking sites for the SH3 and SH2 domains of ITK, respectively.
- the NH2 -terminal region has pro line-containing sequences sharing similarities with the consensus sequence involved in the interaction with the SH3 domain of ITK 36 .
- These sequences in CD27 are highly conserved in mammals. We thus examined whether ITK could functionally or directly interact with CD27. We first tested the capacity of CD27 to trigger tyrosine phosphorylation events.
- Tyrosine phosphorylated proteins were detected in lysates from control T-cell blasts of a healthy donor, 15 minutes after stimulation by anti-CD27 antibody. These phosphorylation signals were in part distinct and delayed compared to the signals observed in response to CD3 stimulation. Interestingly, a protein with the size of ITK was phosphorylated upon CD27 but not CD3 stimulation. Further analysis of signals known to be dependent of ITK 16 , such as phosphorylation of PLC- ⁇ and ERK1/2 showed that both pathways were activated upon CD27 stimulation, albeit less intensively than in response to CD3 stimulation.
- CD70 delivers signals to T cells through CD27 and ITK, those being necessary for T-cell proliferation maintenance and hence execution of the effector programs.
- CD70 deficiency appears to be a phenocopy of the CD27 deficiency.
- Phenotypes of CD27- and CD70-deficient mice are also comparable, and both characterized by diminished antiviral responses 22 ' 25 ' 21 .
- the clinical phenotype associated with ITK-deficiency is also very close to the one of CD27 and CD70 deficiencies 7 .
- CD70-CD27 axis represents a key factor of the immune response to EBV in humans.
- CD70 on EBV -infected B cells provides crucial signals to EBV-specific CTLs for their expansion.
- CD70 is strongly upregulated in activated B cells and EBV-infected B cells.
- CD70 is also constitutively expressed by many EBV- and non EBV-dependent B cell malignancies 24 .
- the peculiar predisposition to EBV infection associated with CD27-, CD70- and ITK-deficiencies likely results from the unique tropism of EBV to B cells and its capacity to induce B-cell proliferation and transformation.
- CD27 behaves as a co stimulatory molecule of TCR-dependent lymphocyte activation 29 .
- the co-signals delivered by CD27 are crucial for sustained T-cell expansion as recently highlighted 37 .
- proliferation associated cosignals downstream of CD27 are strictly dependent on ITK that can bind to the intracytoplasmic tail of CD27 via its SH3 domain. It is well established that ITK is directly involved in TCR signaling, by its ability to activate PLC- ⁇ , Ca 2+ flux and ERK kinases 16 ' 18 .
- CD27 engagement also triggers activation of PLC- ⁇ , Ca 2+ flux and ERK kinases, suggesting a quantitative amplification of the TCR strength by CD27 as suggested 29 ' 37 . More studies are warranted to decipher the exact molecular requirements for ITK in CD27 signaling and how it cooperates with TCR signaling to drive cell division.
- CD70 on B cells is a key player in T-cell immunity towards proliferating B cells
- CD70 is also expressed on other hematopoietic cells including sub populations of activated T cells and DC cells.
- CD27-dependent priming and maintenance of T-cell responses was shown to involve interactions with CD70 expressed on DCs 38 ' 39 .
- the absence of CD70 on DCs contributes to the lack of control of EBV infection in the patient, in particular during the priming phase.
- CD70 on DCs seems not to be absolutely required for immune responses to other pathogens. Partial redundancy with other TNFR family members such as 4- 1BB and OX40 that are expressed by T cells could provide similar signals to T cells as those deliver by CD27, when engaged by their ligand during interactions with DC 29 ' 40 .
- CD70 on B cells act as a semaphore to signal abnormal B cell proliferation to T cells even in the absence of EBV as a trigger of B cell proliferation.
- somatic mutations in CD70 including large deletions have been identified in B-cell lymphomas such as diffuse large B cell lymphomas and Burkitt's lymphomas 42 ' 43 ' 44 .
- a recent report identified CD70 as one of the most frequently mutated genes in a series of diffuse large B-cell lymphomas 45 . Accumulation of mutations in CD70 may represent a mechanism for malignant B cells to escape immune surveillance by T cells.
- CD70-CD27 could represent an important target to induce anti-tumoral vaccination.
- Restoration or induction of CD70 expression in some lymphoma cells lacking CD70 might represent a therapeutic approach to induce a global and potent anti-tumoral immunity.
- Ibrutinib is an irreversible molecular inhibitor of ITK driving a Thl -selective pressure in
- CD27 is a signal-transducing molecule involved in CD45RA+ naive T cell costimulation. Journal of immunology 1994, 153(12): 5422-5432. 34. DeBarros A, Chaves-Ferreira M, d'Orey F, Ribot JC, Silva-Santos B. CD70- CD27 interactions provide survival and proliferative signals that regulate T cell receptor-driven activation of human gammadelta peripheral blood lymphocytes. European journal of immunology 2011, 41(1): 195-201.
- the costimulatory molecule CD70 is regulated by distinct molecular mechanisms and is associated with overall survival in diffuse large B-cell lymphoma. Genes, chromosomes & cancer 2013, 52(8): 764-774. 44. Scholtysik R, Nagel I, Stamm M, Vater I, Giefmg M, Schwaenen C, et al. Recurrent deletions of the TNFSF7 and TNFSF9 genes in 19 l3.3 in diffuse large B-cell and Burkitt lymphomas. International journal of cancer 2012, 131(5): E830-835.
Abstract
Description
Claims
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
EP16305471 | 2016-04-22 | ||
PCT/EP2017/059466 WO2017182608A1 (en) | 2016-04-22 | 2017-04-21 | Methods and vaccine compositions for the treatment of b-cell malignancies |
Publications (1)
Publication Number | Publication Date |
---|---|
EP3445378A1 true EP3445378A1 (en) | 2019-02-27 |
Family
ID=56024206
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
EP17720714.9A Withdrawn EP3445378A1 (en) | 2016-04-22 | 2017-04-21 | Methods and vaccine compositions for the treatment of b-cell malignancies |
Country Status (3)
Country | Link |
---|---|
EP (1) | EP3445378A1 (en) |
JP (1) | JP2019514870A (en) |
WO (1) | WO2017182608A1 (en) |
Families Citing this family (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN111154913B (en) * | 2018-11-08 | 2023-04-18 | 中山大学 | Primers and crRNA for EB virus DNA detection and application thereof |
Family Cites Families (9)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
FR901228A (en) | 1943-01-16 | 1945-07-20 | Deutsche Edelstahlwerke Ag | Ring gap magnet system |
IL85035A0 (en) | 1987-01-08 | 1988-06-30 | Int Genetic Eng | Polynucleotide molecule,a chimeric antibody with specificity for human b cell surface antigen,a process for the preparation and methods utilizing the same |
US5057423A (en) | 1987-12-18 | 1991-10-15 | University Of Pittsburgh | Method for the preparation of pure LAK-active lymphocytes |
US5736137A (en) | 1992-11-13 | 1998-04-07 | Idec Pharmaceuticals Corporation | Therapeutic application of chimeric and radiolabeled antibodies to human B lymphocyte restricted differentiation antigen for treatment of B cell lymphoma |
US5595721A (en) | 1993-09-16 | 1997-01-21 | Coulter Pharmaceutical, Inc. | Radioimmunotherapy of lymphoma using anti-CD20 |
US6013516A (en) | 1995-10-06 | 2000-01-11 | The Salk Institute For Biological Studies | Vector and method of use for nucleic acid delivery to non-dividing cells |
US7321026B2 (en) | 2001-06-27 | 2008-01-22 | Skytech Technology Limited | Framework-patched immunoglobulins |
KR20100050587A (en) | 2002-10-17 | 2010-05-13 | 젠맵 에이/에스 | Human monoclonal antibodies against cd20 |
CN103501816A (en) * | 2010-10-27 | 2014-01-08 | 贝勒医学院 | Chimeric CD27 receptors for redirecting t cells to CD70-positive malignancies |
-
2017
- 2017-04-21 JP JP2018554695A patent/JP2019514870A/en active Pending
- 2017-04-21 WO PCT/EP2017/059466 patent/WO2017182608A1/en active Application Filing
- 2017-04-21 EP EP17720714.9A patent/EP3445378A1/en not_active Withdrawn
Also Published As
Publication number | Publication date |
---|---|
JP2019514870A (en) | 2019-06-06 |
WO2017182608A1 (en) | 2017-10-26 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
Izawa et al. | Inherited CD70 deficiency in humans reveals a critical role for the CD70–CD27 pathway in immunity to Epstein-Barr virus infection | |
EP3823665B1 (en) | Chimeric antigen receptors with bcma specificity and uses thereof | |
AU2017319151B2 (en) | Immune cell compositions and methods of use for treating viral and other infections | |
CN109997041B (en) | Human leukocyte antigen-restricted gamma delta T cell receptors and methods of use thereof | |
JP6856188B2 (en) | Immune effector cells genetically engineered with CS1-specific chimeric antigen receptors | |
US10716838B2 (en) | Anti-CD277 antibodies | |
Takamura et al. | Premature terminal exhaustion of Friend virus-specific effector CD8+ T cells by rapid induction of multiple inhibitory receptors | |
TW202005658A (en) | T cell receptors and engineered cells expressing same | |
JP7447388B2 (en) | Coreceptor systems for the treatment of infectious diseases | |
EP3383419B1 (en) | Compositions and methods for reducing immune responses against chimeric antigen receptors | |
KR20180033537A (en) | PD-L1 expressing hematopoietic stem cells and uses | |
EP3843778A1 (en) | Chimeric antigen receptors against multiple hla-g isoforms | |
US20220281994A1 (en) | Chimeric Antigen Receptors with MAGE-A4 Specificity and Uses Thereof | |
EP3445378A1 (en) | Methods and vaccine compositions for the treatment of b-cell malignancies | |
EP3999540A1 (en) | Antibodies having specificity for cd38 and uses thereof | |
US11904004B2 (en) | Anti-CD277 antibodies | |
Mbanwi | Regulation of anti-viral immunity by dendritic cells and natural killer cells | |
Rose | Influence of NK cells on the anti-influenza immune response | |
Sáez Borderias | Regulation of natural killer and cd4+ T cell function by NKG2 C-type lectin-like receptors | |
Muramatsu et al. | Biology of Blood and Marrow Transplantation |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
STAA | Information on the status of an ep patent application or granted ep patent |
Free format text: STATUS: UNKNOWN |
|
STAA | Information on the status of an ep patent application or granted ep patent |
Free format text: STATUS: THE INTERNATIONAL PUBLICATION HAS BEEN MADE |
|
PUAI | Public reference made under article 153(3) epc to a published international application that has entered the european phase |
Free format text: ORIGINAL CODE: 0009012 |
|
STAA | Information on the status of an ep patent application or granted ep patent |
Free format text: STATUS: REQUEST FOR EXAMINATION WAS MADE |
|
17P | Request for examination filed |
Effective date: 20181018 |
|
AK | Designated contracting states |
Kind code of ref document: A1 Designated state(s): AL AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HR HU IE IS IT LI LT LU LV MC MK MT NL NO PL PT RO RS SE SI SK SM TR |
|
AX | Request for extension of the european patent |
Extension state: BA ME |
|
DAV | Request for validation of the european patent (deleted) | ||
DAX | Request for extension of the european patent (deleted) | ||
STAA | Information on the status of an ep patent application or granted ep patent |
Free format text: STATUS: EXAMINATION IS IN PROGRESS |
|
17Q | First examination report despatched |
Effective date: 20191126 |
|
STAA | Information on the status of an ep patent application or granted ep patent |
Free format text: STATUS: THE APPLICATION IS DEEMED TO BE WITHDRAWN |
|
18D | Application deemed to be withdrawn |
Effective date: 20200609 |