WO2021076887A1 - Cal-t constructs and uses thereof - Google Patents
Cal-t constructs and uses thereof Download PDFInfo
- Publication number
- WO2021076887A1 WO2021076887A1 PCT/US2020/055980 US2020055980W WO2021076887A1 WO 2021076887 A1 WO2021076887 A1 WO 2021076887A1 US 2020055980 W US2020055980 W US 2020055980W WO 2021076887 A1 WO2021076887 A1 WO 2021076887A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- seq
- gliadin
- gamma
- immunogenic
- domain
- Prior art date
Links
- 239000000203 mixture Substances 0.000 claims abstract description 74
- 230000006916 protein interaction Effects 0.000 claims abstract description 72
- 238000000034 method Methods 0.000 claims abstract description 37
- 230000004068 intracellular signaling Effects 0.000 claims abstract description 25
- 208000023275 Autoimmune disease Diseases 0.000 claims abstract description 13
- 206010052779 Transplant rejections Diseases 0.000 claims abstract description 6
- 108090000765 processed proteins & peptides Proteins 0.000 claims description 248
- 230000003993 interaction Effects 0.000 claims description 131
- 102000004196 processed proteins & peptides Human genes 0.000 claims description 107
- 229920001184 polypeptide Polymers 0.000 claims description 100
- 210000004027 cell Anatomy 0.000 claims description 83
- 108700018351 Major Histocompatibility Complex Proteins 0.000 claims description 76
- 230000020382 suppression by virus of host antigen processing and presentation of peptide antigen via MHC class I Effects 0.000 claims description 76
- 210000001744 T-lymphocyte Anatomy 0.000 claims description 65
- 239000000427 antigen Substances 0.000 claims description 55
- 102000036639 antigens Human genes 0.000 claims description 55
- 108091007433 antigens Proteins 0.000 claims description 55
- 108010019670 Chimeric Antigen Receptors Proteins 0.000 claims description 52
- 230000027455 binding Effects 0.000 claims description 52
- 230000001363 autoimmune Effects 0.000 claims description 42
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims description 39
- 102100028972 HLA class I histocompatibility antigen, A alpha chain Human genes 0.000 claims description 35
- 108010075704 HLA-A Antigens Proteins 0.000 claims description 35
- 108010088729 HLA-A*02:01 antigen Proteins 0.000 claims description 33
- 241000282414 Homo sapiens Species 0.000 claims description 28
- 102000018679 Tacrolimus Binding Proteins Human genes 0.000 claims description 28
- 108010027179 Tacrolimus Binding Proteins Proteins 0.000 claims description 28
- 230000001404 mediated effect Effects 0.000 claims description 28
- 201000010099 disease Diseases 0.000 claims description 25
- 230000011664 signaling Effects 0.000 claims description 25
- 108090000623 proteins and genes Proteins 0.000 claims description 23
- 235000018102 proteins Nutrition 0.000 claims description 22
- 102000004169 proteins and genes Human genes 0.000 claims description 22
- 208000011580 syndromic disease Diseases 0.000 claims description 22
- 102100028976 HLA class I histocompatibility antigen, B alpha chain Human genes 0.000 claims description 20
- 108010058607 HLA-B Antigens Proteins 0.000 claims description 20
- 230000001684 chronic effect Effects 0.000 claims description 20
- 206010018364 Glomerulonephritis Diseases 0.000 claims description 18
- 206010003246 arthritis Diseases 0.000 claims description 18
- 206010012601 diabetes mellitus Diseases 0.000 claims description 18
- 230000001154 acute effect Effects 0.000 claims description 16
- 208000010668 atopic eczema Diseases 0.000 claims description 16
- 208000015943 Coeliac disease Diseases 0.000 claims description 15
- 206010067584 Type 1 diabetes mellitus Diseases 0.000 claims description 15
- 230000000172 allergic effect Effects 0.000 claims description 14
- 208000035475 disorder Diseases 0.000 claims description 14
- 230000004927 fusion Effects 0.000 claims description 14
- 206010025135 lupus erythematosus Diseases 0.000 claims description 14
- 206010047642 Vitiligo Diseases 0.000 claims description 13
- 230000000735 allogeneic effect Effects 0.000 claims description 13
- 239000000539 dimer Substances 0.000 claims description 12
- 239000003446 ligand Substances 0.000 claims description 12
- 201000000596 systemic lupus erythematosus Diseases 0.000 claims description 12
- 239000013638 trimer Substances 0.000 claims description 12
- 208000031212 Autoimmune polyendocrinopathy Diseases 0.000 claims description 10
- 229930191978 Gibberellin Natural products 0.000 claims description 10
- 102210024302 HLA-B*0702 Human genes 0.000 claims description 10
- 108010078301 HLA-B*07:02 antigen Proteins 0.000 claims description 10
- 108010090804 Streptavidin Proteins 0.000 claims description 10
- 206010047115 Vasculitis Diseases 0.000 claims description 10
- IXORZMNAPKEEDV-UHFFFAOYSA-N gibberellic acid GA3 Natural products OC(=O)C1C2(C3)CC(=C)C3(O)CCC2C2(C=CC3O)C1C3(C)C(=O)O2 IXORZMNAPKEEDV-UHFFFAOYSA-N 0.000 claims description 10
- 239000003448 gibberellin Substances 0.000 claims description 10
- 201000006417 multiple sclerosis Diseases 0.000 claims description 10
- 239000002773 nucleotide Substances 0.000 claims description 10
- 125000003729 nucleotide group Chemical group 0.000 claims description 10
- 210000000056 organ Anatomy 0.000 claims description 10
- 206010039073 rheumatoid arthritis Diseases 0.000 claims description 10
- 102000043129 MHC class I family Human genes 0.000 claims description 9
- 108091054437 MHC class I family Proteins 0.000 claims description 9
- 102000014914 Carrier Proteins Human genes 0.000 claims description 8
- 208000024869 Goodpasture syndrome Diseases 0.000 claims description 8
- 208000030836 Hashimoto thyroiditis Diseases 0.000 claims description 8
- 208000029523 Interstitial Lung disease Diseases 0.000 claims description 8
- 108091054438 MHC class II family Proteins 0.000 claims description 8
- 102000043131 MHC class II family Human genes 0.000 claims description 8
- 206010034277 Pemphigoid Diseases 0.000 claims description 8
- 206010046851 Uveitis Diseases 0.000 claims description 8
- 208000006673 asthma Diseases 0.000 claims description 8
- 108091008324 binding proteins Proteins 0.000 claims description 8
- 206010009887 colitis Diseases 0.000 claims description 8
- 206010014599 encephalitis Diseases 0.000 claims description 8
- 230000002757 inflammatory effect Effects 0.000 claims description 8
- 201000008383 nephritis Diseases 0.000 claims description 8
- 102210042925 HLA-A*02:01 Human genes 0.000 claims description 7
- 206010001052 Acute respiratory distress syndrome Diseases 0.000 claims description 6
- 102000004631 Calcineurin Human genes 0.000 claims description 6
- 108010042955 Calcineurin Proteins 0.000 claims description 6
- 206010008609 Cholangitis sclerosing Diseases 0.000 claims description 6
- 208000002691 Choroiditis Diseases 0.000 claims description 6
- 208000006344 Churg-Strauss Syndrome Diseases 0.000 claims description 6
- 206010011715 Cyclitis Diseases 0.000 claims description 6
- 201000004624 Dermatitis Diseases 0.000 claims description 6
- 206010012442 Dermatitis contact Diseases 0.000 claims description 6
- 208000018428 Eosinophilic granulomatosis with polyangiitis Diseases 0.000 claims description 6
- 201000005569 Gout Diseases 0.000 claims description 6
- 206010021245 Idiopathic thrombocytopenic purpura Diseases 0.000 claims description 6
- 208000010159 IgA glomerulonephritis Diseases 0.000 claims description 6
- 206010021263 IgA nephropathy Diseases 0.000 claims description 6
- 208000022559 Inflammatory bowel disease Diseases 0.000 claims description 6
- 206010025280 Lymphocytosis Diseases 0.000 claims description 6
- 206010029164 Nephrotic syndrome Diseases 0.000 claims description 6
- 201000011152 Pemphigus Diseases 0.000 claims description 6
- 241000721454 Pemphigus Species 0.000 claims description 6
- 201000004681 Psoriasis Diseases 0.000 claims description 6
- 206010039705 Scleritis Diseases 0.000 claims description 6
- 208000034189 Sclerosis Diseases 0.000 claims description 6
- 201000009594 Systemic Scleroderma Diseases 0.000 claims description 6
- 206010042953 Systemic sclerosis Diseases 0.000 claims description 6
- 208000031981 Thrombocytopenic Idiopathic Purpura Diseases 0.000 claims description 6
- 208000007502 anemia Diseases 0.000 claims description 6
- 201000008937 atopic dermatitis Diseases 0.000 claims description 6
- 201000003710 autoimmune thrombocytopenic purpura Diseases 0.000 claims description 6
- 201000002491 encephalomyelitis Diseases 0.000 claims description 6
- 230000002124 endocrine Effects 0.000 claims description 6
- 201000001155 extrinsic allergic alveolitis Diseases 0.000 claims description 6
- 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 claims description 6
- 238000001415 gene therapy Methods 0.000 claims description 6
- 208000022098 hypersensitivity pneumonitis Diseases 0.000 claims description 6
- 230000028993 immune response Effects 0.000 claims description 6
- 208000027866 inflammatory disease Diseases 0.000 claims description 6
- 201000005328 monoclonal gammopathy of uncertain significance Diseases 0.000 claims description 6
- 239000000178 monomer Substances 0.000 claims description 6
- 210000000822 natural killer cell Anatomy 0.000 claims description 6
- 208000012111 paraneoplastic syndrome Diseases 0.000 claims description 6
- 208000033808 peripheral neuropathy Diseases 0.000 claims description 6
- 208000009954 pyoderma gangrenosum Diseases 0.000 claims description 6
- 208000002574 reactive arthritis Diseases 0.000 claims description 6
- 208000010157 sclerosing cholangitis Diseases 0.000 claims description 6
- 208000017520 skin disease Diseases 0.000 claims description 6
- 101800001415 Bri23 peptide Proteins 0.000 claims description 5
- 101800000655 C-terminal peptide Proteins 0.000 claims description 5
- 102400000107 C-terminal peptide Human genes 0.000 claims description 5
- 206010020751 Hypersensitivity Diseases 0.000 claims description 5
- ROHFNLRQFUQHCH-YFKPBYRVSA-N L-leucine Chemical compound CC(C)C[C@H](N)C(O)=O ROHFNLRQFUQHCH-YFKPBYRVSA-N 0.000 claims description 5
- ROHFNLRQFUQHCH-UHFFFAOYSA-N Leucine Natural products CC(C)CC(N)C(O)=O ROHFNLRQFUQHCH-UHFFFAOYSA-N 0.000 claims description 5
- 101150037674 PMA2 gene Proteins 0.000 claims description 5
- HCHKCACWOHOZIP-UHFFFAOYSA-N Zinc Chemical compound [Zn] HCHKCACWOHOZIP-UHFFFAOYSA-N 0.000 claims description 5
- 208000026935 allergic disease Diseases 0.000 claims description 5
- 230000005784 autoimmunity Effects 0.000 claims description 5
- 125000001475 halogen functional group Chemical group 0.000 claims description 5
- 210000000265 leukocyte Anatomy 0.000 claims description 5
- 230000003211 malignant effect Effects 0.000 claims description 5
- 229910052725 zinc Inorganic materials 0.000 claims description 5
- 239000011701 zinc Substances 0.000 claims description 5
- 208000026872 Addison Disease Diseases 0.000 claims description 4
- 208000032671 Allergic granulomatous angiitis Diseases 0.000 claims description 4
- 201000004384 Alopecia Diseases 0.000 claims description 4
- 206010001889 Alveolitis Diseases 0.000 claims description 4
- 208000035939 Alveolitis allergic Diseases 0.000 claims description 4
- 206010002556 Ankylosing Spondylitis Diseases 0.000 claims description 4
- 208000004300 Atrophic Gastritis Diseases 0.000 claims description 4
- 208000032116 Autoimmune Experimental Encephalomyelitis Diseases 0.000 claims description 4
- 206010003827 Autoimmune hepatitis Diseases 0.000 claims description 4
- 208000009137 Behcet syndrome Diseases 0.000 claims description 4
- 208000006373 Bell palsy Diseases 0.000 claims description 4
- 208000031976 Channelopathies Diseases 0.000 claims description 4
- 208000011231 Crohn disease Diseases 0.000 claims description 4
- 102000004127 Cytokines Human genes 0.000 claims description 4
- 108090000695 Cytokines Proteins 0.000 claims description 4
- 102100039498 Cytotoxic T-lymphocyte protein 4 Human genes 0.000 claims description 4
- 108020004414 DNA Proteins 0.000 claims description 4
- 206010011878 Deafness Diseases 0.000 claims description 4
- 208000006313 Delayed Hypersensitivity Diseases 0.000 claims description 4
- 208000016192 Demyelinating disease Diseases 0.000 claims description 4
- 206010012434 Dermatitis allergic Diseases 0.000 claims description 4
- 206010012438 Dermatitis atopic Diseases 0.000 claims description 4
- 206010051392 Diapedesis Diseases 0.000 claims description 4
- 208000005373 Dyshidrotic Eczema Diseases 0.000 claims description 4
- 206010014950 Eosinophilia Diseases 0.000 claims description 4
- 201000003542 Factor VIII deficiency Diseases 0.000 claims description 4
- 206010018634 Gouty Arthritis Diseases 0.000 claims description 4
- 208000015023 Graves' disease Diseases 0.000 claims description 4
- 208000035895 Guillain-Barré syndrome Diseases 0.000 claims description 4
- 208000035186 Hemolytic Autoimmune Anemia Diseases 0.000 claims description 4
- 208000009292 Hemophilia A Diseases 0.000 claims description 4
- 206010019939 Herpes gestationis Diseases 0.000 claims description 4
- 101000889276 Homo sapiens Cytotoxic T-lymphocyte protein 4 Proteins 0.000 claims description 4
- 101001137987 Homo sapiens Lymphocyte activation gene 3 protein Proteins 0.000 claims description 4
- 101000914514 Homo sapiens T-cell-specific surface glycoprotein CD28 Proteins 0.000 claims description 4
- 208000000038 Hypoparathyroidism Diseases 0.000 claims description 4
- 201000009794 Idiopathic Pulmonary Fibrosis Diseases 0.000 claims description 4
- 206010061218 Inflammation Diseases 0.000 claims description 4
- 206010022941 Iridocyclitis Diseases 0.000 claims description 4
- 102000017578 LAG3 Human genes 0.000 claims description 4
- 201000010743 Lambert-Eaton myasthenic syndrome Diseases 0.000 claims description 4
- 201000001779 Leukocyte adhesion deficiency Diseases 0.000 claims description 4
- 201000009906 Meningitis Diseases 0.000 claims description 4
- 206010049567 Miller Fisher syndrome Diseases 0.000 claims description 4
- 208000003250 Mixed connective tissue disease Diseases 0.000 claims description 4
- 208000010190 Monoclonal Gammopathy of Undetermined Significance Diseases 0.000 claims description 4
- 206010028424 Myasthenic syndrome Diseases 0.000 claims description 4
- 201000002481 Myositis Diseases 0.000 claims description 4
- 206010028665 Myxoedema Diseases 0.000 claims description 4
- 206010029240 Neuritis Diseases 0.000 claims description 4
- 208000005225 Opsoclonus-Myoclonus Syndrome Diseases 0.000 claims description 4
- 208000008223 Pemphigoid Gestationis Diseases 0.000 claims description 4
- 206010065159 Polychondritis Diseases 0.000 claims description 4
- 206010036105 Polyneuropathy Diseases 0.000 claims description 4
- 208000003971 Posterior uveitis Diseases 0.000 claims description 4
- 102100024213 Programmed cell death 1 ligand 2 Human genes 0.000 claims description 4
- 208000012322 Raynaud phenomenon Diseases 0.000 claims description 4
- 208000033464 Reiter syndrome Diseases 0.000 claims description 4
- 208000007400 Relapsing-Remitting Multiple Sclerosis Diseases 0.000 claims description 4
- 206010063837 Reperfusion injury Diseases 0.000 claims description 4
- 208000013616 Respiratory Distress Syndrome Diseases 0.000 claims description 4
- 206010039085 Rhinitis allergic Diseases 0.000 claims description 4
- 206010039710 Scleroderma Diseases 0.000 claims description 4
- 206010040047 Sepsis Diseases 0.000 claims description 4
- 102100029215 Signaling lymphocytic activation molecule Human genes 0.000 claims description 4
- 206010042033 Stevens-Johnson syndrome Diseases 0.000 claims description 4
- 206010072148 Stiff-Person syndrome Diseases 0.000 claims description 4
- 102100027213 T-cell-specific surface glycoprotein CD28 Human genes 0.000 claims description 4
- 108010065917 TOR Serine-Threonine Kinases Proteins 0.000 claims description 4
- 102100028785 Tumor necrosis factor receptor superfamily member 14 Human genes 0.000 claims description 4
- 102100022153 Tumor necrosis factor receptor superfamily member 4 Human genes 0.000 claims description 4
- 208000024780 Urticaria Diseases 0.000 claims description 4
- 206010047124 Vasculitis necrotising Diseases 0.000 claims description 4
- 201000010105 allergic rhinitis Diseases 0.000 claims description 4
- 231100000360 alopecia Toxicity 0.000 claims description 4
- 206010002026 amyotrophic lateral sclerosis Diseases 0.000 claims description 4
- 201000004612 anterior uveitis Diseases 0.000 claims description 4
- 208000002399 aphthous stomatitis Diseases 0.000 claims description 4
- 201000000448 autoimmune hemolytic anemia Diseases 0.000 claims description 4
- 208000027625 autoimmune inner ear disease Diseases 0.000 claims description 4
- 210000003719 b-lymphocyte Anatomy 0.000 claims description 4
- 208000002479 balanitis Diseases 0.000 claims description 4
- 238000002659 cell therapy Methods 0.000 claims description 4
- 210000003169 central nervous system Anatomy 0.000 claims description 4
- 208000016644 chronic atrophic gastritis Diseases 0.000 claims description 4
- 208000025302 chronic primary adrenal insufficiency Diseases 0.000 claims description 4
- 208000024376 chronic urticaria Diseases 0.000 claims description 4
- 208000010247 contact dermatitis Diseases 0.000 claims description 4
- 201000001981 dermatomyositis Diseases 0.000 claims description 4
- 208000030172 endocrine system disease Diseases 0.000 claims description 4
- 206010014801 endophthalmitis Diseases 0.000 claims description 4
- 210000003979 eosinophil Anatomy 0.000 claims description 4
- 201000005206 focal segmental glomerulosclerosis Diseases 0.000 claims description 4
- 231100000854 focal segmental glomerulosclerosis Toxicity 0.000 claims description 4
- 230000010370 hearing loss Effects 0.000 claims description 4
- 231100000888 hearing loss Toxicity 0.000 claims description 4
- 208000016354 hearing loss disease Diseases 0.000 claims description 4
- 208000007475 hemolytic anemia Diseases 0.000 claims description 4
- 208000006454 hepatitis Diseases 0.000 claims description 4
- 231100000283 hepatitis Toxicity 0.000 claims description 4
- 208000003532 hypothyroidism Diseases 0.000 claims description 4
- 206010021198 ichthyosis Diseases 0.000 claims description 4
- 230000008595 infiltration Effects 0.000 claims description 4
- 238000001764 infiltration Methods 0.000 claims description 4
- 230000004054 inflammatory process Effects 0.000 claims description 4
- 208000036971 interstitial lung disease 2 Diseases 0.000 claims description 4
- 201000002364 leukopenia Diseases 0.000 claims description 4
- 201000008350 membranous glomerulonephritis Diseases 0.000 claims description 4
- 208000037890 multiple organ injury Diseases 0.000 claims description 4
- 206010028417 myasthenia gravis Diseases 0.000 claims description 4
- 208000003786 myxedema Diseases 0.000 claims description 4
- 208000008795 neuromyelitis optica Diseases 0.000 claims description 4
- 201000001119 neuropathy Diseases 0.000 claims description 4
- 230000007823 neuropathy Effects 0.000 claims description 4
- 208000005963 oophoritis Diseases 0.000 claims description 4
- 201000006292 polyarteritis nodosa Diseases 0.000 claims description 4
- 230000007824 polyneuropathy Effects 0.000 claims description 4
- 206010063401 primary progressive multiple sclerosis Diseases 0.000 claims description 4
- 201000000742 primary sclerosing cholangitis Diseases 0.000 claims description 4
- 230000002062 proliferating effect Effects 0.000 claims description 4
- 208000005069 pulmonary fibrosis Diseases 0.000 claims description 4
- 230000004044 response Effects 0.000 claims description 4
- 201000000306 sarcoidosis Diseases 0.000 claims description 4
- 206010043778 thyroiditis Diseases 0.000 claims description 4
- 208000035408 type 1 diabetes mellitus 1 Diseases 0.000 claims description 4
- 208000001072 type 2 diabetes mellitus Diseases 0.000 claims description 4
- 210000004369 blood Anatomy 0.000 claims description 3
- 239000008280 blood Substances 0.000 claims description 3
- 210000004443 dendritic cell Anatomy 0.000 claims description 3
- 229940079593 drug Drugs 0.000 claims description 3
- 239000003814 drug Substances 0.000 claims description 3
- 210000001519 tissue Anatomy 0.000 claims description 3
- 102100033051 40S ribosomal protein S19 Human genes 0.000 claims description 2
- XGWFJBFNAQHLEF-UHFFFAOYSA-N 9-anthroic acid Chemical compound C1=CC=C2C(C(=O)O)=C(C=CC=C3)C3=CC2=C1 XGWFJBFNAQHLEF-UHFFFAOYSA-N 0.000 claims description 2
- 206010000748 Acute febrile neutrophilic dermatosis Diseases 0.000 claims description 2
- 206010001076 Acute sinusitis Diseases 0.000 claims description 2
- 206010062269 Adrenalitis Diseases 0.000 claims description 2
- 201000010000 Agranulocytosis Diseases 0.000 claims description 2
- 208000035285 Allergic Seasonal Rhinitis Diseases 0.000 claims description 2
- 206010027654 Allergic conditions Diseases 0.000 claims description 2
- 206010001766 Alopecia totalis Diseases 0.000 claims description 2
- 208000024985 Alport syndrome Diseases 0.000 claims description 2
- 206010001881 Alveolar proteinosis Diseases 0.000 claims description 2
- 208000024827 Alzheimer disease Diseases 0.000 claims description 2
- 206010001935 American trypanosomiasis Diseases 0.000 claims description 2
- 206010002198 Anaphylactic reaction Diseases 0.000 claims description 2
- 206010002412 Angiocentric lymphomas Diseases 0.000 claims description 2
- 208000002267 Anti-neutrophil cytoplasmic antibody-associated vasculitis Diseases 0.000 claims description 2
- 208000003343 Antiphospholipid Syndrome Diseases 0.000 claims description 2
- 206010002961 Aplasia Diseases 0.000 claims description 2
- 206010003210 Arteriosclerosis Diseases 0.000 claims description 2
- 206010003267 Arthritis reactive Diseases 0.000 claims description 2
- 206010003591 Ataxia Diseases 0.000 claims description 2
- 206010003594 Ataxia telangiectasia Diseases 0.000 claims description 2
- 201000001320 Atherosclerosis Diseases 0.000 claims description 2
- 208000012657 Atopic disease Diseases 0.000 claims description 2
- 206010003645 Atopy Diseases 0.000 claims description 2
- 206010003694 Atrophy Diseases 0.000 claims description 2
- 206010003805 Autism Diseases 0.000 claims description 2
- 208000020706 Autistic disease Diseases 0.000 claims description 2
- 206010071576 Autoimmune aplastic anaemia Diseases 0.000 claims description 2
- 206010064539 Autoimmune myocarditis Diseases 0.000 claims description 2
- 206010055128 Autoimmune neutropenia Diseases 0.000 claims description 2
- 208000023095 Autosomal dominant epidermolytic ichthyosis Diseases 0.000 claims description 2
- 102100038080 B-cell receptor CD22 Human genes 0.000 claims description 2
- 206010004078 Balanoposthitis Diseases 0.000 claims description 2
- 208000023328 Basedow disease Diseases 0.000 claims description 2
- 208000027496 Behcet disease Diseases 0.000 claims description 2
- 208000008439 Biliary Liver Cirrhosis Diseases 0.000 claims description 2
- 208000033222 Biliary cirrhosis primary Diseases 0.000 claims description 2
- 208000033932 Blackfan-Diamond anemia Diseases 0.000 claims description 2
- 102100027207 CD27 antigen Human genes 0.000 claims description 2
- 101150013553 CD40 gene Proteins 0.000 claims description 2
- 201000007155 CD40 ligand deficiency Diseases 0.000 claims description 2
- 102100035793 CD83 antigen Human genes 0.000 claims description 2
- 201000002829 CREST Syndrome Diseases 0.000 claims description 2
- 208000004434 Calcinosis Diseases 0.000 claims description 2
- 208000020119 Caplan syndrome Diseases 0.000 claims description 2
- 102100025466 Carcinoembryonic antigen-related cell adhesion molecule 3 Human genes 0.000 claims description 2
- 102100024965 Caspase recruitment domain-containing protein 11 Human genes 0.000 claims description 2
- 208000024699 Chagas disease Diseases 0.000 claims description 2
- 206010008748 Chorea Diseases 0.000 claims description 2
- 206010008874 Chronic Fatigue Syndrome Diseases 0.000 claims description 2
- 208000030939 Chronic inflammatory demyelinating polyneuropathy Diseases 0.000 claims description 2
- 206010009137 Chronic sinusitis Diseases 0.000 claims description 2
- 102100026735 Coagulation factor VIII Human genes 0.000 claims description 2
- 208000010007 Cogan syndrome Diseases 0.000 claims description 2
- 206010009900 Colitis ulcerative Diseases 0.000 claims description 2
- 102000000503 Collagen Type II Human genes 0.000 claims description 2
- 108010041390 Collagen Type II Proteins 0.000 claims description 2
- 208000027932 Collagen disease Diseases 0.000 claims description 2
- 102100030886 Complement receptor type 1 Human genes 0.000 claims description 2
- 206010053138 Congenital aplastic anaemia Diseases 0.000 claims description 2
- 208000014311 Cushing syndrome Diseases 0.000 claims description 2
- 206010011686 Cutaneous vasculitis Diseases 0.000 claims description 2
- 201000003883 Cystic fibrosis Diseases 0.000 claims description 2
- 102000053602 DNA Human genes 0.000 claims description 2
- 241000702421 Dependoparvovirus Species 0.000 claims description 2
- 206010012455 Dermatitis exfoliative Diseases 0.000 claims description 2
- 206010012468 Dermatitis herpetiformis Diseases 0.000 claims description 2
- 208000007342 Diabetic Nephropathies Diseases 0.000 claims description 2
- 206010012689 Diabetic retinopathy Diseases 0.000 claims description 2
- 201000004449 Diamond-Blackfan anemia Diseases 0.000 claims description 2
- 201000003066 Diffuse Scleroderma Diseases 0.000 claims description 2
- 208000006926 Discoid Lupus Erythematosus Diseases 0.000 claims description 2
- 208000021866 Dressler syndrome Diseases 0.000 claims description 2
- 208000003556 Dry Eye Syndromes Diseases 0.000 claims description 2
- 206010013774 Dry eye Diseases 0.000 claims description 2
- 206010014190 Eczema asteatotic Diseases 0.000 claims description 2
- 206010014201 Eczema nummular Diseases 0.000 claims description 2
- 208000037487 Endotoxemia Diseases 0.000 claims description 2
- 208000004232 Enteritis Diseases 0.000 claims description 2
- 206010014952 Eosinophilia myalgia syndrome Diseases 0.000 claims description 2
- 206010014954 Eosinophilic fasciitis Diseases 0.000 claims description 2
- 201000009040 Epidermolytic Hyperkeratosis Diseases 0.000 claims description 2
- 206010015084 Episcleritis Diseases 0.000 claims description 2
- 206010015150 Erythema Diseases 0.000 claims description 2
- 206010015153 Erythema annulare Diseases 0.000 claims description 2
- 206010055035 Erythema dyschromicum perstans Diseases 0.000 claims description 2
- 206010015218 Erythema multiforme Diseases 0.000 claims description 2
- 206010015226 Erythema nodosum Diseases 0.000 claims description 2
- 206010015251 Erythroblastosis foetalis Diseases 0.000 claims description 2
- 208000030644 Esophageal Motility disease Diseases 0.000 claims description 2
- 208000004332 Evans syndrome Diseases 0.000 claims description 2
- 208000009386 Experimental Arthritis Diseases 0.000 claims description 2
- 208000027445 Farmer Lung Diseases 0.000 claims description 2
- 208000028387 Felty syndrome Diseases 0.000 claims description 2
- 208000001640 Fibromyalgia Diseases 0.000 claims description 2
- 206010016654 Fibrosis Diseases 0.000 claims description 2
- 208000036495 Gastritis atrophic Diseases 0.000 claims description 2
- 208000018522 Gastrointestinal disease Diseases 0.000 claims description 2
- 208000007465 Giant cell arteritis Diseases 0.000 claims description 2
- 206010018366 Glomerulonephritis acute Diseases 0.000 claims description 2
- 206010018367 Glomerulonephritis chronic Diseases 0.000 claims description 2
- 206010018372 Glomerulonephritis membranous Diseases 0.000 claims description 2
- 206010018691 Granuloma Diseases 0.000 claims description 2
- 201000005708 Granuloma Annulare Diseases 0.000 claims description 2
- 206010072579 Granulomatosis with polyangiitis Diseases 0.000 claims description 2
- 208000003807 Graves Disease Diseases 0.000 claims description 2
- 208000008899 Habitual abortion Diseases 0.000 claims description 2
- 206010018910 Haemolysis Diseases 0.000 claims description 2
- 208000031071 Hamman-Rich Syndrome Diseases 0.000 claims description 2
- 208000001204 Hashimoto Disease Diseases 0.000 claims description 2
- 102100029360 Hematopoietic cell signal transducer Human genes 0.000 claims description 2
- 208000032843 Hemorrhage Diseases 0.000 claims description 2
- 201000004331 Henoch-Schoenlein purpura Diseases 0.000 claims description 2
- 206010019617 Henoch-Schonlein purpura Diseases 0.000 claims description 2
- 206010019860 Hereditary angioedema Diseases 0.000 claims description 2
- 208000007514 Herpes zoster Diseases 0.000 claims description 2
- 101000884305 Homo sapiens B-cell receptor CD22 Proteins 0.000 claims description 2
- 101000914511 Homo sapiens CD27 antigen Proteins 0.000 claims description 2
- 101000946856 Homo sapiens CD83 antigen Proteins 0.000 claims description 2
- 101000914337 Homo sapiens Carcinoembryonic antigen-related cell adhesion molecule 3 Proteins 0.000 claims description 2
- 101000761179 Homo sapiens Caspase recruitment domain-containing protein 11 Proteins 0.000 claims description 2
- 101000911390 Homo sapiens Coagulation factor VIII Proteins 0.000 claims description 2
- 101000727061 Homo sapiens Complement receptor type 1 Proteins 0.000 claims description 2
- 101000990188 Homo sapiens Hematopoietic cell signal transducer Proteins 0.000 claims description 2
- 101000599852 Homo sapiens Intercellular adhesion molecule 1 Proteins 0.000 claims description 2
- 101000851376 Homo sapiens Tumor necrosis factor receptor superfamily member 8 Proteins 0.000 claims description 2
- 101000818543 Homo sapiens Tyrosine-protein kinase ZAP-70 Proteins 0.000 claims description 2
- 206010020631 Hypergammaglobulinaemia benign monoclonal Diseases 0.000 claims description 2
- 206010020649 Hyperkeratosis Diseases 0.000 claims description 2
- 206010020850 Hyperthyroidism Diseases 0.000 claims description 2
- 206010021067 Hypopituitarism Diseases 0.000 claims description 2
- 208000016300 Idiopathic chronic eosinophilic pneumonia Diseases 0.000 claims description 2
- 208000031814 IgA Vasculitis Diseases 0.000 claims description 2
- 208000004575 Infectious Arthritis Diseases 0.000 claims description 2
- 102100037877 Intercellular adhesion molecule 1 Human genes 0.000 claims description 2
- 206010022557 Intermediate uveitis Diseases 0.000 claims description 2
- 208000000209 Isaacs syndrome Diseases 0.000 claims description 2
- 208000009388 Job Syndrome Diseases 0.000 claims description 2
- 208000012528 Juvenile dermatomyositis Diseases 0.000 claims description 2
- 206010059176 Juvenile idiopathic arthritis Diseases 0.000 claims description 2
- 208000011200 Kawasaki disease Diseases 0.000 claims description 2
- 206010023335 Keratitis interstitial Diseases 0.000 claims description 2
- 208000009319 Keratoconjunctivitis Sicca Diseases 0.000 claims description 2
- 208000001126 Keratosis Diseases 0.000 claims description 2
- 208000001913 Lamellar ichthyosis Diseases 0.000 claims description 2
- 208000034624 Leukocytoclastic Cutaneous Vasculitis Diseases 0.000 claims description 2
- 208000032514 Leukocytoclastic vasculitis Diseases 0.000 claims description 2
- 208000007820 Lichen Sclerosus et Atrophicus Diseases 0.000 claims description 2
- 206010024434 Lichen sclerosus Diseases 0.000 claims description 2
- 206010024436 Lichen spinulosus Diseases 0.000 claims description 2
- 208000001244 Linear IgA Bullous Dermatosis Diseases 0.000 claims description 2
- 208000004883 Lipoid Nephrosis Diseases 0.000 claims description 2
- 201000009324 Loeffler syndrome Diseases 0.000 claims description 2
- 206010025102 Lung infiltration Diseases 0.000 claims description 2
- 208000005777 Lupus Nephritis Diseases 0.000 claims description 2
- 208000016604 Lyme disease Diseases 0.000 claims description 2
- 102100034709 Lymphocyte cytosolic protein 2 Human genes 0.000 claims description 2
- 101710195102 Lymphocyte cytosolic protein 2 Proteins 0.000 claims description 2
- 206010025323 Lymphomas Diseases 0.000 claims description 2
- 108010061593 Member 14 Tumor Necrosis Factor Receptors Proteins 0.000 claims description 2
- 101100407308 Mus musculus Pdcd1lg2 gene Proteins 0.000 claims description 2
- 208000000112 Myalgia Diseases 0.000 claims description 2
- 208000009525 Myocarditis Diseases 0.000 claims description 2
- 206010051606 Necrotising colitis Diseases 0.000 claims description 2
- 206010065673 Nephritic syndrome Diseases 0.000 claims description 2
- 208000028389 Nerve injury Diseases 0.000 claims description 2
- 201000009053 Neurodermatitis Diseases 0.000 claims description 2
- 206010072359 Neuromyotonia Diseases 0.000 claims description 2
- 208000015914 Non-Hodgkin lymphomas Diseases 0.000 claims description 2
- 206010029888 Obliterative bronchiolitis Diseases 0.000 claims description 2
- 208000002193 Pain Diseases 0.000 claims description 2
- 206010033661 Pancytopenia Diseases 0.000 claims description 2
- 208000026433 Pemphigus erythematosus Diseases 0.000 claims description 2
- 208000027086 Pemphigus foliaceus Diseases 0.000 claims description 2
- 206010034620 Peripheral sensory neuropathy Diseases 0.000 claims description 2
- 208000031845 Pernicious anaemia Diseases 0.000 claims description 2
- 206010036030 Polyarthritis Diseases 0.000 claims description 2
- 208000007048 Polymyalgia Rheumatica Diseases 0.000 claims description 2
- 206010036242 Post vaccination syndrome Diseases 0.000 claims description 2
- 208000031732 Post-Lyme Disease Syndrome Diseases 0.000 claims description 2
- 206010036297 Postpartum hypopituitarism Diseases 0.000 claims description 2
- 208000006664 Precursor Cell Lymphoblastic Leukemia-Lymphoma Diseases 0.000 claims description 2
- 206010036631 Presenile dementia Diseases 0.000 claims description 2
- 208000002500 Primary Ovarian Insufficiency Diseases 0.000 claims description 2
- 208000012654 Primary biliary cholangitis Diseases 0.000 claims description 2
- 206010036697 Primary hypothyroidism Diseases 0.000 claims description 2
- 108700030875 Programmed Cell Death 1 Ligand 2 Proteins 0.000 claims description 2
- 102100024216 Programmed cell death 1 ligand 1 Human genes 0.000 claims description 2
- 208000003251 Pruritus Diseases 0.000 claims description 2
- 201000001263 Psoriatic Arthritis Diseases 0.000 claims description 2
- 208000036824 Psoriatic arthropathy Diseases 0.000 claims description 2
- 206010037575 Pustular psoriasis Diseases 0.000 claims description 2
- 206010071141 Rasmussen encephalitis Diseases 0.000 claims description 2
- 208000004160 Rasmussen subacute encephalitis Diseases 0.000 claims description 2
- 208000021329 Refractory celiac disease Diseases 0.000 claims description 2
- 206010038748 Restrictive cardiomyopathy Diseases 0.000 claims description 2
- 208000025747 Rheumatic disease Diseases 0.000 claims description 2
- 241001303601 Rosacea Species 0.000 claims description 2
- 206010039793 Seborrhoeic dermatitis Diseases 0.000 claims description 2
- 206010040070 Septic Shock Diseases 0.000 claims description 2
- 208000032384 Severe immune-mediated enteropathy Diseases 0.000 claims description 2
- 201000009895 Sheehan syndrome Diseases 0.000 claims description 2
- 108010074687 Signaling Lymphocytic Activation Molecule Family Member 1 Proteins 0.000 claims description 2
- 208000021386 Sjogren Syndrome Diseases 0.000 claims description 2
- 231100000168 Stevens-Johnson syndrome Toxicity 0.000 claims description 2
- 206010042342 Subcorneal pustular dermatosis Diseases 0.000 claims description 2
- 206010061373 Sudden Hearing Loss Diseases 0.000 claims description 2
- 208000010265 Sweet syndrome Diseases 0.000 claims description 2
- 208000027522 Sydenham chorea Diseases 0.000 claims description 2
- 208000029052 T-cell acute lymphoblastic leukemia Diseases 0.000 claims description 2
- 102100025237 T-cell surface antigen CD2 Human genes 0.000 claims description 2
- 206010043189 Telangiectasia Diseases 0.000 claims description 2
- 206010043781 Thyroiditis chronic Diseases 0.000 claims description 2
- 206010043784 Thyroiditis subacute Diseases 0.000 claims description 2
- 206010044223 Toxic epidermal necrolysis Diseases 0.000 claims description 2
- 231100000087 Toxic epidermal necrolysis Toxicity 0.000 claims description 2
- 206010044248 Toxic shock syndrome Diseases 0.000 claims description 2
- 231100000650 Toxic shock syndrome Toxicity 0.000 claims description 2
- 206010044314 Tracheobronchitis Diseases 0.000 claims description 2
- 208000003441 Transfusion reaction Diseases 0.000 claims description 2
- 206010051446 Transient acantholytic dermatosis Diseases 0.000 claims description 2
- 108010023649 Tripartite Motif Proteins Proteins 0.000 claims description 2
- 101710165473 Tumor necrosis factor receptor superfamily member 4 Proteins 0.000 claims description 2
- 102100040245 Tumor necrosis factor receptor superfamily member 5 Human genes 0.000 claims description 2
- 102100036857 Tumor necrosis factor receptor superfamily member 8 Human genes 0.000 claims description 2
- 208000006391 Type 1 Hyper-IgM Immunodeficiency Syndrome Diseases 0.000 claims description 2
- 206010070517 Type 2 lepra reaction Diseases 0.000 claims description 2
- 102100021125 Tyrosine-protein kinase ZAP-70 Human genes 0.000 claims description 2
- 201000006704 Ulcerative Colitis Diseases 0.000 claims description 2
- 208000036826 VIIth nerve paralysis Diseases 0.000 claims description 2
- 206010047112 Vasculitides Diseases 0.000 claims description 2
- 102100026383 Vasopressin-neurophysin 2-copeptin Human genes 0.000 claims description 2
- 208000014926 Vesiculobullous Skin disease Diseases 0.000 claims description 2
- 208000006110 Wiskott-Aldrich syndrome Diseases 0.000 claims description 2
- 201000001696 X-linked hyper IgM syndrome Diseases 0.000 claims description 2
- 208000037855 acute anterior uveitis Diseases 0.000 claims description 2
- 208000026816 acute arthritis Diseases 0.000 claims description 2
- 231100000851 acute glomerulonephritis Toxicity 0.000 claims description 2
- 201000004073 acute interstitial pneumonia Diseases 0.000 claims description 2
- 208000011341 adult acute respiratory distress syndrome Diseases 0.000 claims description 2
- 201000000028 adult respiratory distress syndrome Diseases 0.000 claims description 2
- 208000002029 allergic contact dermatitis Diseases 0.000 claims description 2
- 208000030961 allergic reaction Diseases 0.000 claims description 2
- 201000010435 allergic urticaria Diseases 0.000 claims description 2
- 208000004631 alopecia areata Diseases 0.000 claims description 2
- 206010002022 amyloidosis Diseases 0.000 claims description 2
- 230000036783 anaphylactic response Effects 0.000 claims description 2
- 208000003455 anaphylaxis Diseases 0.000 claims description 2
- 208000011775 arteriosclerosis disease Diseases 0.000 claims description 2
- 206010003230 arteritis Diseases 0.000 claims description 2
- 230000001977 ataxic effect Effects 0.000 claims description 2
- 230000037444 atrophy Effects 0.000 claims description 2
- 208000001974 autoimmune enteropathy Diseases 0.000 claims description 2
- 201000004982 autoimmune uveitis Diseases 0.000 claims description 2
- 201000000751 autosomal recessive congenital ichthyosis Diseases 0.000 claims description 2
- 208000015440 bird fancier lung Diseases 0.000 claims description 2
- 230000008993 bowel inflammation Effects 0.000 claims description 2
- 210000000133 brain stem Anatomy 0.000 claims description 2
- 201000003848 bronchiolitis obliterans Diseases 0.000 claims description 2
- 208000023367 bronchiolitis obliterans with obstructive pulmonary disease Diseases 0.000 claims description 2
- 208000010353 central nervous system vasculitis Diseases 0.000 claims description 2
- 208000025434 cerebellar degeneration Diseases 0.000 claims description 2
- 201000008191 cerebritis Diseases 0.000 claims description 2
- 239000003795 chemical substances by application Substances 0.000 claims description 2
- 201000010415 childhood type dermatomyositis Diseases 0.000 claims description 2
- 201000004709 chorioretinitis Diseases 0.000 claims description 2
- 201000009323 chronic eosinophilic pneumonia Diseases 0.000 claims description 2
- 208000030949 chronic idiopathic urticaria Diseases 0.000 claims description 2
- 201000005795 chronic inflammatory demyelinating polyneuritis Diseases 0.000 claims description 2
- 230000012085 chronic inflammatory response Effects 0.000 claims description 2
- 208000027157 chronic rhinosinusitis Diseases 0.000 claims description 2
- 206010072757 chronic spontaneous urticaria Diseases 0.000 claims description 2
- 230000007882 cirrhosis Effects 0.000 claims description 2
- 208000019425 cirrhosis of liver Diseases 0.000 claims description 2
- 208000008609 collagenous colitis Diseases 0.000 claims description 2
- 235000009508 confectionery Nutrition 0.000 claims description 2
- 208000029078 coronary artery disease Diseases 0.000 claims description 2
- 210000003792 cranial nerve Anatomy 0.000 claims description 2
- 201000003278 cryoglobulinemia Diseases 0.000 claims description 2
- 208000004921 cutaneous lupus erythematosus Diseases 0.000 claims description 2
- 201000010064 diabetes insipidus Diseases 0.000 claims description 2
- 208000033679 diabetic kidney disease Diseases 0.000 claims description 2
- 208000032625 disorder of ear Diseases 0.000 claims description 2
- 201000011191 dyskinesia of esophagus Diseases 0.000 claims description 2
- 210000003162 effector t lymphocyte Anatomy 0.000 claims description 2
- 206010014665 endocarditis Diseases 0.000 claims description 2
- 201000010048 endomyocardial fibrosis Diseases 0.000 claims description 2
- 238000002641 enzyme replacement therapy Methods 0.000 claims description 2
- 230000002327 eosinophilic effect Effects 0.000 claims description 2
- 208000033286 epidermolytic ichthyosis Diseases 0.000 claims description 2
- 231100000321 erythema Toxicity 0.000 claims description 2
- 201000009320 ethmoid sinusitis Diseases 0.000 claims description 2
- 208000004526 exfoliative dermatitis Diseases 0.000 claims description 2
- 208000022195 farmer lung disease Diseases 0.000 claims description 2
- 230000001605 fetal effect Effects 0.000 claims description 2
- 208000001031 fetal erythroblastosis Diseases 0.000 claims description 2
- 201000006916 frontal sinusitis Diseases 0.000 claims description 2
- ZZUFCTLCJUWOSV-UHFFFAOYSA-N furosemide Chemical compound C1=C(Cl)C(S(=O)(=O)N)=CC(C(O)=O)=C1NCC1=CC=CO1 ZZUFCTLCJUWOSV-UHFFFAOYSA-N 0.000 claims description 2
- 238000010362 genome editing Methods 0.000 claims description 2
- 230000002710 gonadal effect Effects 0.000 claims description 2
- 206010018797 guttate psoriasis Diseases 0.000 claims description 2
- 208000014951 hematologic disease Diseases 0.000 claims description 2
- 210000003958 hematopoietic stem cell Anatomy 0.000 claims description 2
- 230000008588 hemolysis Effects 0.000 claims description 2
- 208000003215 hereditary nephritis Diseases 0.000 claims description 2
- 210000005260 human cell Anatomy 0.000 claims description 2
- 208000014796 hyper-IgE recurrent infection syndrome 1 Diseases 0.000 claims description 2
- 206010051040 hyper-IgE syndrome Diseases 0.000 claims description 2
- 208000026095 hyper-IgM syndrome type 1 Diseases 0.000 claims description 2
- 230000003463 hyperproliferative effect Effects 0.000 claims description 2
- 230000009610 hypersensitivity Effects 0.000 claims description 2
- 201000006362 hypersensitivity vasculitis Diseases 0.000 claims description 2
- 230000002989 hypothyroidism Effects 0.000 claims description 2
- 208000013397 idiopathic acute eosinophilic pneumonia Diseases 0.000 claims description 2
- 230000001900 immune effect Effects 0.000 claims description 2
- 230000008105 immune reaction Effects 0.000 claims description 2
- 208000015446 immunoglobulin a vasculitis Diseases 0.000 claims description 2
- 208000000509 infertility Diseases 0.000 claims description 2
- 230000036512 infertility Effects 0.000 claims description 2
- 231100000535 infertility Toxicity 0.000 claims description 2
- 208000030603 inherited susceptibility to asthma Diseases 0.000 claims description 2
- 208000014674 injury Diseases 0.000 claims description 2
- 201000006904 interstitial keratitis Diseases 0.000 claims description 2
- 201000004614 iritis Diseases 0.000 claims description 2
- 208000001875 irritant dermatitis Diseases 0.000 claims description 2
- 208000012947 ischemia reperfusion injury Diseases 0.000 claims description 2
- 230000000302 ischemic effect Effects 0.000 claims description 2
- 230000000366 juvenile effect Effects 0.000 claims description 2
- 201000002215 juvenile rheumatoid arthritis Diseases 0.000 claims description 2
- 201000010901 lateral sclerosis Diseases 0.000 claims description 2
- 230000021633 leukocyte mediated immunity Effects 0.000 claims description 2
- 231100001022 leukopenia Toxicity 0.000 claims description 2
- 206010024428 lichen nitidus Diseases 0.000 claims description 2
- 201000011486 lichen planus Diseases 0.000 claims description 2
- 230000002197 limbic effect Effects 0.000 claims description 2
- 208000029631 linear IgA Dermatosis Diseases 0.000 claims description 2
- 201000011649 lymphoblastic lymphoma Diseases 0.000 claims description 2
- 230000000527 lymphocytic effect Effects 0.000 claims description 2
- 208000006116 lymphomatoid granulomatosis Diseases 0.000 claims description 2
- 201000008836 maxillary sinusitis Diseases 0.000 claims description 2
- 239000012528 membrane Substances 0.000 claims description 2
- 231100000855 membranous nephropathy Toxicity 0.000 claims description 2
- 208000008275 microscopic colitis Diseases 0.000 claims description 2
- 206010063344 microscopic polyangiitis Diseases 0.000 claims description 2
- 208000005264 motor neuron disease Diseases 0.000 claims description 2
- 208000001725 mucocutaneous lymph node syndrome Diseases 0.000 claims description 2
- 206010065579 multifocal motor neuropathy Diseases 0.000 claims description 2
- 208000029766 myalgic encephalomeyelitis/chronic fatigue syndrome Diseases 0.000 claims description 2
- 208000004995 necrotizing enterocolitis Diseases 0.000 claims description 2
- 230000008764 nerve damage Effects 0.000 claims description 2
- 208000002040 neurosyphilis Diseases 0.000 claims description 2
- 201000004071 non-specific interstitial pneumonia Diseases 0.000 claims description 2
- 208000028780 ocular motility disease Diseases 0.000 claims description 2
- 201000005737 orchitis Diseases 0.000 claims description 2
- 201000008482 osteoarthritis Diseases 0.000 claims description 2
- 210000001672 ovary Anatomy 0.000 claims description 2
- 201000001976 pemphigus vulgaris Diseases 0.000 claims description 2
- 201000006195 perinatal necrotizing enterocolitis Diseases 0.000 claims description 2
- 208000030428 polyarticular arthritis Diseases 0.000 claims description 2
- 208000005987 polymyositis Diseases 0.000 claims description 2
- 208000006473 polyradiculopathy Diseases 0.000 claims description 2
- 230000035935 pregnancy Effects 0.000 claims description 2
- 206010036601 premature menopause Diseases 0.000 claims description 2
- 208000017942 premature ovarian failure 1 Diseases 0.000 claims description 2
- 201000009395 primary hyperaldosteronism Diseases 0.000 claims description 2
- 230000000750 progressive effect Effects 0.000 claims description 2
- 201000003489 pulmonary alveolar proteinosis Diseases 0.000 claims description 2
- 230000002685 pulmonary effect Effects 0.000 claims description 2
- 201000009732 pulmonary eosinophilia Diseases 0.000 claims description 2
- 206010061928 radiculitis Diseases 0.000 claims description 2
- 210000003289 regulatory T cell Anatomy 0.000 claims description 2
- 208000009169 relapsing polychondritis Diseases 0.000 claims description 2
- 230000010410 reperfusion Effects 0.000 claims description 2
- 230000002207 retinal effect Effects 0.000 claims description 2
- 201000003068 rheumatic fever Diseases 0.000 claims description 2
- 201000004700 rosacea Diseases 0.000 claims description 2
- 210000003786 sclera Anatomy 0.000 claims description 2
- 208000008742 seborrheic dermatitis Diseases 0.000 claims description 2
- 201000005572 sensory peripheral neuropathy Diseases 0.000 claims description 2
- 201000001223 septic arthritis Diseases 0.000 claims description 2
- 208000013223 septicemia Diseases 0.000 claims description 2
- 201000009890 sinusitis Diseases 0.000 claims description 2
- 239000007787 solid Substances 0.000 claims description 2
- 201000006923 sphenoid sinusitis Diseases 0.000 claims description 2
- 201000007497 subacute thyroiditis Diseases 0.000 claims description 2
- 201000004595 synovitis Diseases 0.000 claims description 2
- 208000002025 tabes dorsalis Diseases 0.000 claims description 2
- 208000009056 telangiectasis Diseases 0.000 claims description 2
- 206010043207 temporal arteritis Diseases 0.000 claims description 2
- 210000001550 testis Anatomy 0.000 claims description 2
- 206010043554 thrombocytopenia Diseases 0.000 claims description 2
- 208000008732 thymoma Diseases 0.000 claims description 2
- 208000005057 thyrotoxicosis Diseases 0.000 claims description 2
- 208000016367 transient hypogammaglobulinemia of infancy Diseases 0.000 claims description 2
- 238000002054 transplantation Methods 0.000 claims description 2
- 208000009174 transverse myelitis Diseases 0.000 claims description 2
- 230000008733 trauma Effects 0.000 claims description 2
- 210000001745 uvea Anatomy 0.000 claims description 2
- 102000013530 TOR Serine-Threonine Kinases Human genes 0.000 claims 2
- 238000005516 engineering process Methods 0.000 abstract description 7
- 208000009329 Graft vs Host Disease Diseases 0.000 abstract description 4
- 208000024908 graft versus host disease Diseases 0.000 abstract description 4
- 102210047469 A*02:01 Human genes 0.000 description 700
- 230000002163 immunogen Effects 0.000 description 609
- 101710087459 Gamma-gliadin Proteins 0.000 description 397
- 108010061711 Gliadin Proteins 0.000 description 270
- 108010050792 glutenin Proteins 0.000 description 138
- 108091008874 T cell receptors Proteins 0.000 description 110
- 102000016266 T-Cell Antigen Receptors Human genes 0.000 description 110
- 241000209140 Triticum Species 0.000 description 78
- 235000021307 Triticum Nutrition 0.000 description 78
- 102210048101 A*01:01 Human genes 0.000 description 60
- 102210047218 B*07:02 Human genes 0.000 description 54
- 208000001490 Dengue Diseases 0.000 description 52
- 206010012310 Dengue fever Diseases 0.000 description 52
- 208000025729 dengue disease Diseases 0.000 description 52
- 102210048112 DRB1*04:01 Human genes 0.000 description 51
- 101710183587 Omega-gliadin Proteins 0.000 description 49
- 102210042961 A*03:01 Human genes 0.000 description 46
- 241000713772 Human immunodeficiency virus 1 Species 0.000 description 36
- 102100030086 Receptor tyrosine-protein kinase erbB-2 Human genes 0.000 description 34
- 208000037797 influenza A Diseases 0.000 description 31
- 102210048109 DRB1*01:01 Human genes 0.000 description 30
- 241000725303 Human immunodeficiency virus Species 0.000 description 30
- 102210048102 B*08:01 Human genes 0.000 description 28
- 102000003425 Tyrosinase Human genes 0.000 description 23
- 108060008724 Tyrosinase Proteins 0.000 description 23
- 102210048104 B*27:05 Human genes 0.000 description 22
- 101710132601 Capsid protein Proteins 0.000 description 22
- 241000701024 Human betaherpesvirus 5 Species 0.000 description 22
- 239000002243 precursor Substances 0.000 description 22
- 241000701022 Cytomegalovirus Species 0.000 description 21
- 102210047482 DRB1*07:01 Human genes 0.000 description 21
- 101001012157 Homo sapiens Receptor tyrosine-protein kinase erbB-2 Proteins 0.000 description 18
- 101800001271 Surface protein Proteins 0.000 description 18
- 102210047362 DRB1*15:01 Human genes 0.000 description 16
- 101710120463 Prostate stem cell antigen Proteins 0.000 description 16
- 102100036735 Prostate stem cell antigen Human genes 0.000 description 16
- 101710100968 Receptor tyrosine-protein kinase erbB-2 Proteins 0.000 description 16
- 231100000331 toxic Toxicity 0.000 description 15
- 230000002588 toxic effect Effects 0.000 description 15
- 102210047483 DRB1*11:01 Human genes 0.000 description 14
- 101001005719 Homo sapiens Melanoma-associated antigen 3 Proteins 0.000 description 14
- 102100025082 Melanoma-associated antigen 3 Human genes 0.000 description 14
- 102100025570 Cancer/testis antigen 1 Human genes 0.000 description 13
- 101000856237 Homo sapiens Cancer/testis antigen 1 Proteins 0.000 description 13
- 239000004698 Polyethylene Substances 0.000 description 13
- 108010069621 Epstein-Barr virus EBV-associated membrane antigen Proteins 0.000 description 12
- 101001131990 Homo sapiens Peroxidasin homolog Proteins 0.000 description 12
- 108700020134 Human immunodeficiency virus 1 nef Proteins 0.000 description 12
- 206010028980 Neoplasm Diseases 0.000 description 12
- 102100034601 Peroxidasin homolog Human genes 0.000 description 12
- 108010076039 Polyproteins Proteins 0.000 description 12
- 108010017842 Telomerase Proteins 0.000 description 12
- 241001678559 COVID-19 virus Species 0.000 description 11
- 102100041003 Glutamate carboxypeptidase 2 Human genes 0.000 description 11
- 101000629313 Severe acute respiratory syndrome coronavirus Spike glycoprotein Proteins 0.000 description 11
- 108700039791 Hepatitis C virus nucleocapsid Proteins 0.000 description 10
- 108091006197 SARS-CoV-2 Nucleocapsid Protein Proteins 0.000 description 10
- 102100032938 Telomerase reverse transcriptase Human genes 0.000 description 10
- AEMBWNDIEFEPTH-UHFFFAOYSA-N n-tert-butyl-n-ethylnitrous amide Chemical compound CCN(N=O)C(C)(C)C AEMBWNDIEFEPTH-UHFFFAOYSA-N 0.000 description 10
- 108700028369 Alleles Proteins 0.000 description 9
- 101150078891 BRLF1 gene Proteins 0.000 description 9
- 101000892862 Homo sapiens Glutamate carboxypeptidase 2 Proteins 0.000 description 9
- 101001100327 Homo sapiens RNA-binding protein 45 Proteins 0.000 description 9
- 102100038823 RNA-binding protein 45 Human genes 0.000 description 9
- 230000004913 activation Effects 0.000 description 9
- 229920001481 poly(stearyl methacrylate) Polymers 0.000 description 9
- WEVYNIUIFUYDGI-UHFFFAOYSA-N 3-[6-[4-(trifluoromethoxy)anilino]-4-pyrimidinyl]benzamide Chemical compound NC(=O)C1=CC=CC(C=2N=CN=C(NC=3C=CC(OC(F)(F)F)=CC=3)C=2)=C1 WEVYNIUIFUYDGI-UHFFFAOYSA-N 0.000 description 8
- 102100021569 Apoptosis regulator Bcl-2 Human genes 0.000 description 8
- 102100040901 Circadian clock protein PASD1 Human genes 0.000 description 8
- 102210047356 DRB1*03:01 Human genes 0.000 description 8
- 102000017177 Fibromodulin Human genes 0.000 description 8
- 108010013996 Fibromodulin Proteins 0.000 description 8
- 101000971171 Homo sapiens Apoptosis regulator Bcl-2 Proteins 0.000 description 8
- 102100031413 L-dopachrome tautomerase Human genes 0.000 description 8
- 101710093778 L-dopachrome tautomerase Proteins 0.000 description 8
- 108060006580 PRAME Proteins 0.000 description 8
- 102000036673 PRAME Human genes 0.000 description 8
- 239000004237 Ponceau 6R Substances 0.000 description 8
- 102100035703 Prostatic acid phosphatase Human genes 0.000 description 8
- 101000629318 Severe acute respiratory syndrome coronavirus 2 Spike glycoprotein Proteins 0.000 description 8
- 241000710886 West Nile virus Species 0.000 description 8
- 239000001752 chlorophylls and chlorophyllins Substances 0.000 description 8
- 239000001679 citrus red 2 Substances 0.000 description 8
- 210000002307 prostate Anatomy 0.000 description 8
- 102000005962 receptors Human genes 0.000 description 8
- 108020003175 receptors Proteins 0.000 description 8
- 229940122450 Altered peptide ligand Drugs 0.000 description 7
- 102100025137 Early activation antigen CD69 Human genes 0.000 description 7
- 101000934374 Homo sapiens Early activation antigen CD69 Proteins 0.000 description 7
- 240000005979 Hordeum vulgare Species 0.000 description 7
- 235000007340 Hordeum vulgare Nutrition 0.000 description 7
- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 description 7
- 239000004472 Lysine Substances 0.000 description 7
- 108020001507 fusion proteins Proteins 0.000 description 7
- 102000037865 fusion proteins Human genes 0.000 description 7
- 230000000670 limiting effect Effects 0.000 description 7
- 125000003588 lysine group Chemical group [H]N([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])(N([H])[H])C(*)=O 0.000 description 7
- SSOORFWOBGFTHL-OTEJMHTDSA-N (4S)-5-[[(2S)-1-[[(2S)-1-[[(2S)-1-[[(2S)-1-[[(2S)-1-[[(2S)-1-[[(2S)-1-[[(2S)-6-amino-1-[[(2S)-1-[[(2S)-1-[[(2S)-1-[[(2S)-1-[[2-[(2S)-2-[[(2S)-1-[[(2S)-1-[[(2S)-1-[[(2S)-1-[[(2S)-1-[[(2S)-1-[[(2S)-6-amino-1-[[(2S)-1-[[(2S)-1-[[(2S,3S)-1-[[(2S)-1-[[(2S)-1-[[(2S)-6-amino-1-[[(2S)-1-[[(2S)-1-[[(2S)-1-[[(2S)-1-[[(2S)-1-[[(2S)-5-amino-1-[[(2S)-1-[[(2S)-1-[[(2S)-6-amino-1-[[(2S)-6-amino-1-[[(2S)-1-[[(2S)-1-[[(2S)-5-amino-1-[[(2S)-5-carbamimidamido-1-[[(2S)-5-carbamimidamido-1-[[(1S)-4-carbamimidamido-1-carboxybutyl]amino]-1-oxopentan-2-yl]amino]-1-oxopentan-2-yl]amino]-1,5-dioxopentan-2-yl]amino]-5-carbamimidamido-1-oxopentan-2-yl]amino]-5-carbamimidamido-1-oxopentan-2-yl]amino]-1-oxohexan-2-yl]amino]-1-oxohexan-2-yl]amino]-5-carbamimidamido-1-oxopentan-2-yl]amino]-4-methyl-1-oxopentan-2-yl]amino]-1,5-dioxopentan-2-yl]amino]-4-methyl-1-oxopentan-2-yl]amino]-3-hydroxy-1-oxopropan-2-yl]amino]-3-hydroxy-1-oxopropan-2-yl]amino]-3-hydroxy-1-oxopropan-2-yl]amino]-1-oxopropan-2-yl]amino]-1-oxohexan-2-yl]amino]-3-hydroxy-1-oxopropan-2-yl]amino]-1-oxo-3-phenylpropan-2-yl]amino]-3-methyl-1-oxopentan-2-yl]amino]-3-methyl-1-oxobutan-2-yl]amino]-5-carbamimidamido-1-oxopentan-2-yl]amino]-1-oxohexan-2-yl]amino]-3-methyl-1-oxobutan-2-yl]amino]-5-carbamimidamido-1-oxopentan-2-yl]amino]-3-methyl-1-oxobutan-2-yl]amino]-4-methyl-1-oxopentan-2-yl]amino]-1-oxopropan-2-yl]amino]-5-carbamimidamido-1-oxopentan-2-yl]carbamoyl]pyrrolidin-1-yl]-2-oxoethyl]amino]-3-(1H-indol-3-yl)-1-oxopropan-2-yl]amino]-4-methyl-1-oxopentan-2-yl]amino]-1-oxo-3-phenylpropan-2-yl]amino]-5-carbamimidamido-1-oxopentan-2-yl]amino]-1-oxohexan-2-yl]amino]-3-methyl-1-oxobutan-2-yl]amino]-5-carbamimidamido-1-oxopentan-2-yl]amino]-4-methyl-1-oxopentan-2-yl]amino]-1-oxo-3-phenylpropan-2-yl]amino]-3-(1H-imidazol-4-yl)-1-oxopropan-2-yl]amino]-3-methyl-1-oxobutan-2-yl]amino]-4-methyl-1-oxopentan-2-yl]amino]-4-[[(2S)-2-[[(2S)-2-[[(2S)-2,6-diaminohexanoyl]amino]-3-methylbutanoyl]amino]propanoyl]amino]-5-oxopentanoic acid Chemical compound CC[C@H](C)[C@H](NC(=O)[C@@H](NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@H](CCCCN)NC(=O)[C@@H](NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@@H](NC(=O)[C@H](CC(C)C)NC(=O)[C@H](C)NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@@H]1CCCN1C(=O)CNC(=O)[C@H](Cc1c[nH]c2ccccc12)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](Cc1ccccc1)NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@H](CCCCN)NC(=O)[C@@H](NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](Cc1ccccc1)NC(=O)[C@H](Cc1c[nH]cn1)NC(=O)[C@@H](NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](C)NC(=O)[C@@H](NC(=O)[C@@H](N)CCCCN)C(C)C)C(C)C)C(C)C)C(C)C)C(C)C)C(C)C)C(=O)N[C@@H](Cc1ccccc1)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C)C(=O)N[C@@H](CO)C(=O)N[C@@H](CO)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O SSOORFWOBGFTHL-OTEJMHTDSA-N 0.000 description 6
- 108700026758 Adenovirus hexon capsid Proteins 0.000 description 6
- 101710145634 Antigen 1 Proteins 0.000 description 6
- 101150062763 BMRF1 gene Proteins 0.000 description 6
- 102100021663 Baculoviral IAP repeat-containing protein 5 Human genes 0.000 description 6
- 108010001572 Basic-Leucine Zipper Transcription Factors Proteins 0.000 description 6
- 102000000806 Basic-Leucine Zipper Transcription Factors Human genes 0.000 description 6
- 102100026245 E3 ubiquitin-protein ligase RNF43 Human genes 0.000 description 6
- 108010031111 EBV-encoded nuclear antigen 1 Proteins 0.000 description 6
- 101710122231 Epstein-Barr nuclear antigen 3 Proteins 0.000 description 6
- 108010058643 Fungal Proteins Proteins 0.000 description 6
- 108700024845 Hepatitis B virus P Proteins 0.000 description 6
- 101000613559 Homo sapiens Circadian clock protein PASD1 Proteins 0.000 description 6
- 101000692702 Homo sapiens E3 ubiquitin-protein ligase RNF43 Proteins 0.000 description 6
- 101001056180 Homo sapiens Induced myeloid leukemia cell differentiation protein Mcl-1 Proteins 0.000 description 6
- 101001088879 Homo sapiens Lysine-specific demethylase 5D Proteins 0.000 description 6
- 101001005718 Homo sapiens Melanoma-associated antigen 2 Proteins 0.000 description 6
- 101001057156 Homo sapiens Melanoma-associated antigen C2 Proteins 0.000 description 6
- 101001136981 Homo sapiens Proteasome subunit beta type-9 Proteins 0.000 description 6
- 108010084873 Human Immunodeficiency Virus nef Gene Products Proteins 0.000 description 6
- 102100026539 Induced myeloid leukemia cell differentiation protein Mcl-1 Human genes 0.000 description 6
- 102100025081 Melanoma-associated antigen 2 Human genes 0.000 description 6
- 102100027252 Melanoma-associated antigen C2 Human genes 0.000 description 6
- 102000003735 Mesothelin Human genes 0.000 description 6
- 108090000015 Mesothelin Proteins 0.000 description 6
- 241000746983 Phleum pratense Species 0.000 description 6
- 108010076181 Proinsulin Proteins 0.000 description 6
- 102100035764 Proteasome subunit beta type-9 Human genes 0.000 description 6
- 102100034091 Receptor-type tyrosine-protein phosphatase-like N Human genes 0.000 description 6
- 108010002687 Survivin Proteins 0.000 description 6
- 208000035896 Twin-reversed arterial perfusion sequence Diseases 0.000 description 6
- 102000015736 beta 2-Microglobulin Human genes 0.000 description 6
- 108010081355 beta 2-Microglobulin Proteins 0.000 description 6
- 230000000711 cancerogenic effect Effects 0.000 description 6
- 231100000315 carcinogenic Toxicity 0.000 description 6
- 231100000135 cytotoxicity Toxicity 0.000 description 6
- 230000003013 cytotoxicity Effects 0.000 description 6
- 230000000694 effects Effects 0.000 description 6
- 108700008776 hepatitis C virus NS-5 Proteins 0.000 description 6
- 108010043671 prostatic acid phosphatase Proteins 0.000 description 6
- 108010077805 Bacterial Proteins Proteins 0.000 description 5
- 101000989950 Otolemur crassicaudatus Hemoglobin subunit alpha-A Proteins 0.000 description 5
- 230000000875 corresponding effect Effects 0.000 description 5
- 230000014509 gene expression Effects 0.000 description 5
- 229960004641 rituximab Drugs 0.000 description 5
- POVNCJSPYFCWJR-USZUGGBUSA-N (4s)-4-[[(2s)-2-[[(2s)-2-amino-3-(4-hydroxyphenyl)propanoyl]amino]-4-methylpentanoyl]amino]-5-[(2s)-2-[[2-[(2s)-2-[[(2s)-1-[[(2s,3r)-1-[[(1s)-1-carboxy-2-methylpropyl]amino]-3-hydroxy-1-oxobutan-2-yl]amino]-3-methyl-1-oxobutan-2-yl]carbamoyl]pyrrolidin-1- Chemical compound C([C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(=O)N1[C@@H](CCC1)C(=O)NCC(=O)N1[C@@H](CCC1)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C(C)C)C(O)=O)C1=CC=C(O)C=C1 POVNCJSPYFCWJR-USZUGGBUSA-N 0.000 description 4
- KISWVXRQTGLFGD-UHFFFAOYSA-N 2-[[2-[[6-amino-2-[[2-[[2-[[5-amino-2-[[2-[[1-[2-[[6-amino-2-[(2,5-diamino-5-oxopentanoyl)amino]hexanoyl]amino]-5-(diaminomethylideneamino)pentanoyl]pyrrolidine-2-carbonyl]amino]-3-hydroxypropanoyl]amino]-5-oxopentanoyl]amino]-5-(diaminomethylideneamino)p Chemical compound C1CCN(C(=O)C(CCCN=C(N)N)NC(=O)C(CCCCN)NC(=O)C(N)CCC(N)=O)C1C(=O)NC(CO)C(=O)NC(CCC(N)=O)C(=O)NC(CCCN=C(N)N)C(=O)NC(CO)C(=O)NC(CCCCN)C(=O)NC(C(=O)NC(CC(C)C)C(O)=O)CC1=CC=C(O)C=C1 KISWVXRQTGLFGD-UHFFFAOYSA-N 0.000 description 4
- LKKMLIBUAXYLOY-UHFFFAOYSA-N 3-Amino-1-methyl-5H-pyrido[4,3-b]indole Chemical compound N1C2=CC=CC=C2C2=C1C=C(N)N=C2C LKKMLIBUAXYLOY-UHFFFAOYSA-N 0.000 description 4
- 102100039510 Cancer/testis antigen 2 Human genes 0.000 description 4
- 102100025475 Carcinoembryonic antigen-related cell adhesion molecule 5 Human genes 0.000 description 4
- 102100034357 Casein kinase I isoform alpha Human genes 0.000 description 4
- 102100039361 Chondrosarcoma-associated gene 2/3 protein Human genes 0.000 description 4
- 108010015742 Cytochrome P-450 Enzyme System Proteins 0.000 description 4
- 102000003849 Cytochrome P450 Human genes 0.000 description 4
- 102100031695 DnaJ homolog subfamily C member 2 Human genes 0.000 description 4
- 101150059079 EBNA1 gene Proteins 0.000 description 4
- 102100030340 Ephrin type-A receptor 2 Human genes 0.000 description 4
- 102100036725 Epithelial discoidin domain-containing receptor 1 Human genes 0.000 description 4
- 101710131668 Epithelial discoidin domain-containing receptor 1 Proteins 0.000 description 4
- 101710122229 Epstein-Barr nuclear antigen 6 Proteins 0.000 description 4
- 102100036255 Glucose-6-phosphatase 2 Human genes 0.000 description 4
- 108010068370 Glutens Proteins 0.000 description 4
- 102000003886 Glycoproteins Human genes 0.000 description 4
- 108090000288 Glycoproteins Proteins 0.000 description 4
- 108010091373 HA-1 antigen Proteins 0.000 description 4
- 101150051208 HSPH1 gene Proteins 0.000 description 4
- 102100031624 Heat shock protein 105 kDa Human genes 0.000 description 4
- 102100024025 Heparanase Human genes 0.000 description 4
- 102100028818 Heterogeneous nuclear ribonucleoprotein L Human genes 0.000 description 4
- 108010084674 Heterogeneous-Nuclear Ribonucleoprotein L Proteins 0.000 description 4
- 102100038970 Histone-lysine N-methyltransferase EZH2 Human genes 0.000 description 4
- 101000889345 Homo sapiens Cancer/testis antigen 2 Proteins 0.000 description 4
- 101000994700 Homo sapiens Casein kinase I isoform alpha Proteins 0.000 description 4
- 101000745414 Homo sapiens Chondrosarcoma-associated gene 2/3 protein Proteins 0.000 description 4
- 101000845887 Homo sapiens DnaJ homolog subfamily C member 2 Proteins 0.000 description 4
- 101000930907 Homo sapiens Glucose-6-phosphatase 2 Proteins 0.000 description 4
- 101000882127 Homo sapiens Histone-lysine N-methyltransferase EZH2 Proteins 0.000 description 4
- 101000628547 Homo sapiens Metalloreductase STEAP1 Proteins 0.000 description 4
- 101001109719 Homo sapiens Nucleophosmin Proteins 0.000 description 4
- 101000872170 Homo sapiens Polycomb complex protein BMI-1 Proteins 0.000 description 4
- 101000880770 Homo sapiens Protein SSX2 Proteins 0.000 description 4
- 101000984753 Homo sapiens Serine/threonine-protein kinase B-raf Proteins 0.000 description 4
- 101001002197 Homoeomma sp. Mu-theraphotoxin-Hsp1a Proteins 0.000 description 4
- 241000598436 Human T-cell lymphotropic virus Species 0.000 description 4
- 241000714260 Human T-lymphotropic virus 1 Species 0.000 description 4
- 241001135569 Human adenovirus 5 Species 0.000 description 4
- 102000004877 Insulin Human genes 0.000 description 4
- 108090001061 Insulin Proteins 0.000 description 4
- 102100025092 Insulin receptor substrate 2 Human genes 0.000 description 4
- 102100039648 Lactadherin Human genes 0.000 description 4
- 101710128836 Large T antigen Proteins 0.000 description 4
- 241000712899 Lymphocytic choriomeningitis mammarenavirus Species 0.000 description 4
- 101710119980 Macrophage migration inhibitory factor Proteins 0.000 description 4
- 101710104939 Macrophage migration inhibitory factor homolog Proteins 0.000 description 4
- 102100026712 Metalloreductase STEAP1 Human genes 0.000 description 4
- NTIZESTWPVYFNL-UHFFFAOYSA-N Methyl isobutyl ketone Chemical compound CC(C)CC(C)=O NTIZESTWPVYFNL-UHFFFAOYSA-N 0.000 description 4
- 102100034263 Mucin-2 Human genes 0.000 description 4
- 108010008705 Mucin-2 Proteins 0.000 description 4
- 108010063954 Mucins Proteins 0.000 description 4
- 102000015728 Mucins Human genes 0.000 description 4
- 241000186366 Mycobacterium bovis Species 0.000 description 4
- 102000047918 Myelin Basic Human genes 0.000 description 4
- 101710107068 Myelin basic protein Proteins 0.000 description 4
- 102100022678 Nucleophosmin Human genes 0.000 description 4
- 102100033566 Polycomb complex protein BMI-1 Human genes 0.000 description 4
- 108010029485 Protein Isoforms Proteins 0.000 description 4
- 102000001708 Protein Isoforms Human genes 0.000 description 4
- 102100037686 Protein SSX2 Human genes 0.000 description 4
- 108010081208 RMFPNAPYL Proteins 0.000 description 4
- 239000004231 Riboflavin-5-Sodium Phosphate Substances 0.000 description 4
- 102100027103 Serine/threonine-protein kinase B-raf Human genes 0.000 description 4
- 101000779242 Severe acute respiratory syndrome coronavirus 2 ORF3a protein Proteins 0.000 description 4
- 101001110297 Severe acute respiratory syndrome coronavirus 2 Replicase polyprotein 1ab Proteins 0.000 description 4
- 102100033579 Trophoblast glycoprotein Human genes 0.000 description 4
- 208000008383 Wilms tumor Diseases 0.000 description 4
- 102000013529 alpha-Fetoproteins Human genes 0.000 description 4
- 108010026331 alpha-Fetoproteins Proteins 0.000 description 4
- 201000011510 cancer Diseases 0.000 description 4
- 230000002538 fungal effect Effects 0.000 description 4
- 108010037536 heparanase Proteins 0.000 description 4
- NOESYZHRGYRDHS-UHFFFAOYSA-N insulin Chemical compound N1C(=O)C(NC(=O)C(CCC(N)=O)NC(=O)C(CCC(O)=O)NC(=O)C(C(C)C)NC(=O)C(NC(=O)CN)C(C)CC)CSSCC(C(NC(CO)C(=O)NC(CC(C)C)C(=O)NC(CC=2C=CC(O)=CC=2)C(=O)NC(CCC(N)=O)C(=O)NC(CC(C)C)C(=O)NC(CCC(O)=O)C(=O)NC(CC(N)=O)C(=O)NC(CC=2C=CC(O)=CC=2)C(=O)NC(CSSCC(NC(=O)C(C(C)C)NC(=O)C(CC(C)C)NC(=O)C(CC=2C=CC(O)=CC=2)NC(=O)C(CC(C)C)NC(=O)C(C)NC(=O)C(CCC(O)=O)NC(=O)C(C(C)C)NC(=O)C(CC(C)C)NC(=O)C(CC=2NC=NC=2)NC(=O)C(CO)NC(=O)CNC2=O)C(=O)NCC(=O)NC(CCC(O)=O)C(=O)NC(CCCNC(N)=N)C(=O)NCC(=O)NC(CC=3C=CC=CC=3)C(=O)NC(CC=3C=CC=CC=3)C(=O)NC(CC=3C=CC(O)=CC=3)C(=O)NC(C(C)O)C(=O)N3C(CCC3)C(=O)NC(CCCCN)C(=O)NC(C)C(O)=O)C(=O)NC(CC(N)=O)C(O)=O)=O)NC(=O)C(C(C)CC)NC(=O)C(CO)NC(=O)C(C(C)O)NC(=O)C1CSSCC2NC(=O)C(CC(C)C)NC(=O)C(NC(=O)C(CCC(N)=O)NC(=O)C(CC(N)=O)NC(=O)C(NC(=O)C(N)CC=1C=CC=CC=1)C(C)C)CC1=CN=CN1 NOESYZHRGYRDHS-UHFFFAOYSA-N 0.000 description 4
- 210000002752 melanocyte Anatomy 0.000 description 4
- 201000008026 nephroblastoma Diseases 0.000 description 4
- 235000017060 Arachis glabrata Nutrition 0.000 description 3
- 244000105624 Arachis hypogaea Species 0.000 description 3
- 235000010777 Arachis hypogaea Nutrition 0.000 description 3
- 235000018262 Arachis monticola Nutrition 0.000 description 3
- 101150009389 BZLF1 gene Proteins 0.000 description 3
- 241000219495 Betulaceae Species 0.000 description 3
- 108091008048 CMVpp65 Proteins 0.000 description 3
- 108091007741 Chimeric antigen receptor T cells Proteins 0.000 description 3
- 101100347633 Drosophila melanogaster Mhc gene Proteins 0.000 description 3
- 101150113929 EBNA2 gene Proteins 0.000 description 3
- 101000578784 Homo sapiens Melanoma antigen recognized by T-cells 1 Proteins 0.000 description 3
- 102000036770 Islet Amyloid Polypeptide Human genes 0.000 description 3
- 108010041872 Islet Amyloid Polypeptide Proteins 0.000 description 3
- 102100033143 Lysine-specific demethylase 5D Human genes 0.000 description 3
- 102100028389 Melanoma antigen recognized by T-cells 1 Human genes 0.000 description 3
- 101000870363 Oryctolagus cuniculus Glutathione S-transferase Yc Proteins 0.000 description 3
- 101001035650 Otolemur crassicaudatus Hemoglobin subunit alpha-B Proteins 0.000 description 3
- 230000006044 T cell activation Effects 0.000 description 3
- 239000013575 birch pollen allergen Substances 0.000 description 3
- 238000013461 design Methods 0.000 description 3
- 231100000673 dose–response relationship Toxicity 0.000 description 3
- 239000000428 dust Substances 0.000 description 3
- 108010065889 glycyl-leucyl-cysteinyl-threonyl-leucyl-valyl-alanyl-methionyl-leucine Proteins 0.000 description 3
- 102000043390 human KDM5D Human genes 0.000 description 3
- 230000001939 inductive effect Effects 0.000 description 3
- 239000003550 marker Substances 0.000 description 3
- 235000020232 peanut Nutrition 0.000 description 3
- 239000004405 propyl p-hydroxybenzoate Substances 0.000 description 3
- 239000004307 sodium orthophenyl phenol Substances 0.000 description 3
- AHOKKYCUWBLDST-QYULHYBRSA-N (2s)-2-[[(2s)-2-[[(2s)-2-[[(2s)-2-[[(2s)-2-[[2-[[(2s)-2-[[(2s,3s)-2-[[(2s)-2,6-diaminohexanoyl]amino]-3-methylpentanoyl]amino]-3-phenylpropanoyl]amino]acetyl]amino]-3-hydroxypropanoyl]amino]-4-methylpentanoyl]amino]propanoyl]amino]-3-phenylpropanoyl]amino Chemical compound C([C@H](NC(=O)[C@@H](NC(=O)[C@@H](N)CCCCN)[C@@H](C)CC)C(=O)NCC(=O)N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C)C(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N[C@@H](CC(C)C)C(O)=O)C1=CC=CC=C1 AHOKKYCUWBLDST-QYULHYBRSA-N 0.000 description 2
- 101710166488 6 kDa early secretory antigenic target Proteins 0.000 description 2
- PCFGFYKGPMQDBX-FEKONODYSA-N 78355-50-7 Chemical compound C([C@@H](C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](C(C)C)C(=O)NCC(=O)N[C@@H](CCCNC(N)=N)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)NCC(=O)NCC(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N[C@@H](CC(C)C)C(O)=O)NC(=O)CNC(=O)CNC(=O)[C@@H](N)CC=1C=CC(O)=CC=1)C1=CC=CC=C1 PCFGFYKGPMQDBX-FEKONODYSA-N 0.000 description 2
- 108010005465 AC133 Antigen Proteins 0.000 description 2
- 102000005908 AC133 Antigen Human genes 0.000 description 2
- 102100033793 ALK tyrosine kinase receptor Human genes 0.000 description 2
- 101710168331 ALK tyrosine kinase receptor Proteins 0.000 description 2
- 101710190443 Acetyl-CoA carboxylase 1 Proteins 0.000 description 2
- 241000702423 Adeno-associated virus - 2 Species 0.000 description 2
- 102100032959 Alpha-actinin-4 Human genes 0.000 description 2
- 108010049777 Ankyrins Proteins 0.000 description 2
- 102000008102 Ankyrins Human genes 0.000 description 2
- 241001225321 Aspergillus fumigatus Species 0.000 description 2
- 102100022005 B-lymphocyte antigen CD20 Human genes 0.000 description 2
- 102100027522 Baculoviral IAP repeat-containing protein 7 Human genes 0.000 description 2
- 101710177963 Baculoviral IAP repeat-containing protein 7 Proteins 0.000 description 2
- 102100026596 Bcl-2-like protein 1 Human genes 0.000 description 2
- 102100021334 Bcl-2-related protein A1 Human genes 0.000 description 2
- 101100439426 Bradyrhizobium diazoefficiens (strain JCM 10833 / BCRC 13528 / IAM 13628 / NBRC 14792 / USDA 110) groEL4 gene Proteins 0.000 description 2
- 206010006187 Breast cancer Diseases 0.000 description 2
- 208000026310 Breast neoplasm Diseases 0.000 description 2
- 108010077333 CAP1-6D Proteins 0.000 description 2
- 102100022002 CD59 glycoprotein Human genes 0.000 description 2
- 101710176679 CD59 glycoprotein Proteins 0.000 description 2
- 108091033409 CRISPR Proteins 0.000 description 2
- 102100024153 Cadherin-15 Human genes 0.000 description 2
- 241000282836 Camelus dromedarius Species 0.000 description 2
- 241000222120 Candida <Saccharomycetales> Species 0.000 description 2
- 108010022366 Carcinoembryonic Antigen Proteins 0.000 description 2
- 102100026622 Cartilage intermediate layer protein 1 Human genes 0.000 description 2
- 101000963801 Chlamydia trachomatis Major outer membrane porin, serovar A Proteins 0.000 description 2
- 101000963800 Chlamydia trachomatis Major outer membrane porin, serovar B Proteins 0.000 description 2
- 101000963805 Chlamydia trachomatis Major outer membrane porin, serovar C Proteins 0.000 description 2
- 101000963803 Chlamydia trachomatis Major outer membrane porin, serovar E Proteins 0.000 description 2
- 101000963802 Chlamydia trachomatis Major outer membrane porin, serovar F Proteins 0.000 description 2
- 101000963799 Chlamydia trachomatis Major outer membrane porin, serovar H Proteins 0.000 description 2
- 101000963797 Chlamydia trachomatis Major outer membrane porin, serovar L1 Proteins 0.000 description 2
- 101000591958 Chlamydia trachomatis Major outer membrane porin, serovar L3 Proteins 0.000 description 2
- 101800004542 Chondromodulin-1 Proteins 0.000 description 2
- 102400000676 Chondromodulin-1 Human genes 0.000 description 2
- 101710091108 Circadian clock protein PASD1 Proteins 0.000 description 2
- 102000013701 Cyclin-Dependent Kinase 4 Human genes 0.000 description 2
- 108010025464 Cyclin-Dependent Kinase 4 Proteins 0.000 description 2
- 102100033270 Cyclin-dependent kinase inhibitor 1 Human genes 0.000 description 2
- 101710157567 Cyclin-dependent kinase inhibitor 1 Proteins 0.000 description 2
- 102000012466 Cytochrome P450 1B1 Human genes 0.000 description 2
- 108050002014 Cytochrome P450 1B1 Proteins 0.000 description 2
- 102100030074 Dickkopf-related protein 1 Human genes 0.000 description 2
- 101710099518 Dickkopf-related protein 1 Proteins 0.000 description 2
- 102100036109 Dual specificity protein kinase TTK Human genes 0.000 description 2
- 101150076616 EPHA2 gene Proteins 0.000 description 2
- 102100039577 ETS translocation variant 5 Human genes 0.000 description 2
- 108010055196 EphA2 Receptor Proteins 0.000 description 2
- 101710122233 Epstein-Barr nuclear antigen 4 Proteins 0.000 description 2
- 208000006168 Ewing Sarcoma Diseases 0.000 description 2
- 108010003471 Fetal Proteins Proteins 0.000 description 2
- 102000004641 Fetal Proteins Human genes 0.000 description 2
- 102100027844 Fibroblast growth factor receptor 4 Human genes 0.000 description 2
- 108010008599 Forkhead Box Protein M1 Proteins 0.000 description 2
- 102100023374 Forkhead box protein M1 Human genes 0.000 description 2
- 102100035427 Forkhead box protein O1 Human genes 0.000 description 2
- 102100024016 G patch domain and ankyrin repeat-containing protein 1 Human genes 0.000 description 2
- 102100039788 GTPase NRas Human genes 0.000 description 2
- 108010012015 GVYDGREHTV Proteins 0.000 description 2
- 102100032530 Glypican-3 Human genes 0.000 description 2
- 102100031493 Growth arrest-specific protein 7 Human genes 0.000 description 2
- 102100039317 HAUS augmin-like complex subunit 3 Human genes 0.000 description 2
- 108010065192 HER2p63 peptide Proteins 0.000 description 2
- 102100028971 HLA class I histocompatibility antigen, C alpha chain Human genes 0.000 description 2
- 102100031547 HLA class II histocompatibility antigen, DO alpha chain Human genes 0.000 description 2
- 108010074032 HLA-A2 Antigen Proteins 0.000 description 2
- 102000025850 HLA-A2 Antigen Human genes 0.000 description 2
- 108010052199 HLA-C Antigens Proteins 0.000 description 2
- 102000006354 HLA-DR Antigens Human genes 0.000 description 2
- 108010058597 HLA-DR Antigens Proteins 0.000 description 2
- 101710113864 Heat shock protein 90 Proteins 0.000 description 2
- 102100034051 Heat shock protein HSP 90-alpha Human genes 0.000 description 2
- 108010018924 Heme Oxygenase-1 Proteins 0.000 description 2
- 102000002737 Heme Oxygenase-1 Human genes 0.000 description 2
- 102100028006 Heme oxygenase 1 Human genes 0.000 description 2
- 108700008783 Hepatitis C virus E1 Proteins 0.000 description 2
- 102100022623 Hepatocyte growth factor receptor Human genes 0.000 description 2
- 101000797282 Homo sapiens Alpha-actinin-4 Proteins 0.000 description 2
- 101000897405 Homo sapiens B-lymphocyte antigen CD20 Proteins 0.000 description 2
- 101000762242 Homo sapiens Cadherin-15 Proteins 0.000 description 2
- 101000714553 Homo sapiens Cadherin-3 Proteins 0.000 description 2
- 101000914324 Homo sapiens Carcinoembryonic antigen-related cell adhesion molecule 5 Proteins 0.000 description 2
- 101000913767 Homo sapiens Cartilage intermediate layer protein 1 Proteins 0.000 description 2
- 101000659223 Homo sapiens Dual specificity protein kinase TTK Proteins 0.000 description 2
- 101000813745 Homo sapiens ETS translocation variant 5 Proteins 0.000 description 2
- 101000917134 Homo sapiens Fibroblast growth factor receptor 4 Proteins 0.000 description 2
- 101000877727 Homo sapiens Forkhead box protein O1 Proteins 0.000 description 2
- 101000904261 Homo sapiens G patch domain and ankyrin repeat-containing protein 1 Proteins 0.000 description 2
- 101000744505 Homo sapiens GTPase NRas Proteins 0.000 description 2
- 101001014668 Homo sapiens Glypican-3 Proteins 0.000 description 2
- 101000923044 Homo sapiens Growth arrest-specific protein 7 Proteins 0.000 description 2
- 101001035819 Homo sapiens HAUS augmin-like complex subunit 3 Proteins 0.000 description 2
- 101000866278 Homo sapiens HLA class II histocompatibility antigen, DO alpha chain Proteins 0.000 description 2
- 101001079623 Homo sapiens Heme oxygenase 1 Proteins 0.000 description 2
- 101100232357 Homo sapiens IL13RA1 gene Proteins 0.000 description 2
- 101100232360 Homo sapiens IL13RA2 gene Proteins 0.000 description 2
- 101000997670 Homo sapiens Integrin beta-8 Proteins 0.000 description 2
- 101001027621 Homo sapiens Kinesin-like protein KIF20A Proteins 0.000 description 2
- 101001034314 Homo sapiens Lactadherin Proteins 0.000 description 2
- 101000984710 Homo sapiens Lymphocyte-specific protein 1 Proteins 0.000 description 2
- 101001005728 Homo sapiens Melanoma-associated antigen 1 Proteins 0.000 description 2
- 101001005717 Homo sapiens Melanoma-associated antigen 12 Proteins 0.000 description 2
- 101001005724 Homo sapiens Melanoma-associated antigen 9 Proteins 0.000 description 2
- 101001036406 Homo sapiens Melanoma-associated antigen C1 Proteins 0.000 description 2
- 101000934338 Homo sapiens Myeloid cell surface antigen CD33 Proteins 0.000 description 2
- 101000613490 Homo sapiens Paired box protein Pax-3 Proteins 0.000 description 2
- 101000601724 Homo sapiens Paired box protein Pax-5 Proteins 0.000 description 2
- 101000760646 Homo sapiens Phosphatidylserine lipase ABHD16A Proteins 0.000 description 2
- 101001001802 Homo sapiens Pleckstrin homology domain-containing family M member 2 Proteins 0.000 description 2
- 101001001272 Homo sapiens Prostatic acid phosphatase Proteins 0.000 description 2
- 101000928535 Homo sapiens Protein delta homolog 1 Proteins 0.000 description 2
- 101000877404 Homo sapiens Protein enabled homolog Proteins 0.000 description 2
- 101000932478 Homo sapiens Receptor-type tyrosine-protein kinase FLT3 Proteins 0.000 description 2
- 101000700733 Homo sapiens Serine/arginine-rich splicing factor 8 Proteins 0.000 description 2
- 101000897669 Homo sapiens Small RNA 2'-O-methyltransferase Proteins 0.000 description 2
- 101000873927 Homo sapiens Squamous cell carcinoma antigen recognized by T-cells 3 Proteins 0.000 description 2
- 101000825086 Homo sapiens Transcription factor SOX-11 Proteins 0.000 description 2
- 101000851018 Homo sapiens Vascular endothelial growth factor receptor 1 Proteins 0.000 description 2
- 108700020147 Human immunodeficiency virus 1 vif Proteins 0.000 description 2
- 101000622260 Human papillomavirus 33 Protein E7 Proteins 0.000 description 2
- 101100540311 Human papillomavirus type 16 E6 gene Proteins 0.000 description 2
- 108010034219 Insulin Receptor Substrate Proteins Proteins 0.000 description 2
- 101710201820 Insulin receptor substrate 2 Proteins 0.000 description 2
- 101710179420 Insulin receptor substrate 2-B Proteins 0.000 description 2
- 102100033336 Integrin beta-8 Human genes 0.000 description 2
- 101710104185 Interferon antagonist C7 Proteins 0.000 description 2
- 108010074328 Interferon-gamma Proteins 0.000 description 2
- 102000008070 Interferon-gamma Human genes 0.000 description 2
- 102100020791 Interleukin-13 receptor subunit alpha-1 Human genes 0.000 description 2
- 102100037694 Kinesin-like protein KIF20A Human genes 0.000 description 2
- ONIBWKKTOPOVIA-BYPYZUCNSA-N L-Proline Chemical compound OC(=O)[C@@H]1CCCN1 ONIBWKKTOPOVIA-BYPYZUCNSA-N 0.000 description 2
- 101710191666 Lactadherin Proteins 0.000 description 2
- 101710192602 Latent membrane protein 1 Proteins 0.000 description 2
- 102100027105 Lymphocyte-specific protein 1 Human genes 0.000 description 2
- 108010059255 MAGE-A10 antigen Proteins 0.000 description 2
- 108010010995 MART-1 Antigen Proteins 0.000 description 2
- 102000016200 MART-1 Antigen Human genes 0.000 description 2
- 101710105759 Major outer membrane porin Proteins 0.000 description 2
- 101710164702 Major outer membrane protein Proteins 0.000 description 2
- 101710176004 Major viral transcription factor ICP4 homolog Proteins 0.000 description 2
- 102100024299 Maternal embryonic leucine zipper kinase Human genes 0.000 description 2
- 101710154611 Maternal embryonic leucine zipper kinase Proteins 0.000 description 2
- 102100025050 Melanoma-associated antigen 1 Human genes 0.000 description 2
- 102100025084 Melanoma-associated antigen 12 Human genes 0.000 description 2
- 102100025079 Melanoma-associated antigen 9 Human genes 0.000 description 2
- 102100039447 Melanoma-associated antigen C1 Human genes 0.000 description 2
- 241001465754 Metazoa Species 0.000 description 2
- 102000016776 Midkine Human genes 0.000 description 2
- 108010092801 Midkine Proteins 0.000 description 2
- 101800003181 Minor histocompatibility antigen HA-1 Proteins 0.000 description 2
- 102400000047 Minor histocompatibility antigen HA-1 Human genes 0.000 description 2
- 108010008707 Mucin-1 Proteins 0.000 description 2
- 102100034256 Mucin-1 Human genes 0.000 description 2
- 102100034681 Myeloblastin Human genes 0.000 description 2
- 108090000973 Myeloblastin Proteins 0.000 description 2
- 102100025243 Myeloid cell surface antigen CD33 Human genes 0.000 description 2
- 102000003505 Myosin Human genes 0.000 description 2
- 108060008487 Myosin Proteins 0.000 description 2
- 102100038938 Myosin-9 Human genes 0.000 description 2
- 101710204108 Myosin-9 Proteins 0.000 description 2
- ICTPRLMOGAKCOZ-HWVMREAWSA-N NCCCC[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@H](C(=O)N[C@H](C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C(C)C)C(O)=O)C(C)C)CC1=CC=C(O)C=C1 Chemical compound NCCCC[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@H](C(=O)N[C@H](C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C(C)C)C(O)=O)C(C)C)CC1=CC=C(O)C=C1 ICTPRLMOGAKCOZ-HWVMREAWSA-N 0.000 description 2
- 101800001019 Non-structural protein 4B Proteins 0.000 description 2
- 241001263478 Norovirus Species 0.000 description 2
- 108700020796 Oncogene Proteins 0.000 description 2
- 102000004067 Osteocalcin Human genes 0.000 description 2
- 108090000573 Osteocalcin Proteins 0.000 description 2
- 102100037603 P2X purinoceptor 5 Human genes 0.000 description 2
- 101710189969 P2X purinoceptor 5 Proteins 0.000 description 2
- 102100040891 Paired box protein Pax-3 Human genes 0.000 description 2
- 102100037504 Paired box protein Pax-5 Human genes 0.000 description 2
- 241000223960 Plasmodium falciparum Species 0.000 description 2
- 102100036246 Pleckstrin homology domain-containing family M member 2 Human genes 0.000 description 2
- 239000004236 Ponceau SX Substances 0.000 description 2
- 239000004285 Potassium sulphite Substances 0.000 description 2
- 101710154245 Probable ATP-dependent Clp protease ATP-binding subunit Proteins 0.000 description 2
- ONIBWKKTOPOVIA-UHFFFAOYSA-N Proline Natural products OC(=O)C1CCCN1 ONIBWKKTOPOVIA-UHFFFAOYSA-N 0.000 description 2
- 108010072866 Prostate-Specific Antigen Proteins 0.000 description 2
- 102100038358 Prostate-specific antigen Human genes 0.000 description 2
- 102100036467 Protein delta homolog 1 Human genes 0.000 description 2
- 108010089836 Proto-Oncogene Proteins c-met Proteins 0.000 description 2
- 101710104378 Putative malate oxidoreductase [NAD] Proteins 0.000 description 2
- 102100029981 Receptor tyrosine-protein kinase erbB-4 Human genes 0.000 description 2
- 101710100963 Receptor tyrosine-protein kinase erbB-4 Proteins 0.000 description 2
- 102100020718 Receptor-type tyrosine-protein kinase FLT3 Human genes 0.000 description 2
- 208000006265 Renal cell carcinoma Diseases 0.000 description 2
- 102100029289 Serine/arginine-rich splicing factor 8 Human genes 0.000 description 2
- 102100023085 Serine/threonine-protein kinase mTOR Human genes 0.000 description 2
- 101001024647 Severe acute respiratory syndrome coronavirus Nucleoprotein Proteins 0.000 description 2
- 101710173694 Short transient receptor potential channel 2 Proteins 0.000 description 2
- 102100021887 Small RNA 2'-O-methyltransferase Human genes 0.000 description 2
- 239000004280 Sodium formate Substances 0.000 description 2
- 102100035748 Squamous cell carcinoma antigen recognized by T-cells 3 Human genes 0.000 description 2
- 241000193996 Streptococcus pyogenes Species 0.000 description 2
- 102100033920 Synemin Human genes 0.000 description 2
- 102100033455 TGF-beta receptor type-2 Human genes 0.000 description 2
- 101710084188 TGF-beta receptor type-2 Proteins 0.000 description 2
- 102000003610 TRPM8 Human genes 0.000 description 2
- 102100024548 Tensin-3 Human genes 0.000 description 2
- 101710100619 Tensin-3 Proteins 0.000 description 2
- 108010022394 Threonine synthase Proteins 0.000 description 2
- 102000005497 Thymidylate Synthase Human genes 0.000 description 2
- 101710183280 Topoisomerase Proteins 0.000 description 2
- 102100022415 Transcription factor SOX-11 Human genes 0.000 description 2
- 101800001690 Transmembrane protein gp41 Proteins 0.000 description 2
- 101710190034 Trophoblast glycoprotein Proteins 0.000 description 2
- 101150111302 Trpm8 gene Proteins 0.000 description 2
- 108090000704 Tubulin Proteins 0.000 description 2
- 102000004243 Tubulin Human genes 0.000 description 2
- 102100033254 Tumor suppressor ARF Human genes 0.000 description 2
- 108010090473 UDP-N-acetylglucosamine-peptide beta-N-acetylglucosaminyltransferase Proteins 0.000 description 2
- 241000700618 Vaccinia virus Species 0.000 description 2
- 241000700622 Vaccinia virus Copenhagen Species 0.000 description 2
- 102100033178 Vascular endothelial growth factor receptor 1 Human genes 0.000 description 2
- 102000003970 Vinculin Human genes 0.000 description 2
- 108090000384 Vinculin Proteins 0.000 description 2
- 101710201961 Virion infectivity factor Proteins 0.000 description 2
- 101500019620 West Nile virus Non-structural protein 4B Proteins 0.000 description 2
- 108010015542 alpha2-gliadin (56-88) Proteins 0.000 description 2
- 239000004178 amaranth Substances 0.000 description 2
- 229940091771 aspergillus fumigatus Drugs 0.000 description 2
- 108700000711 bcl-X Proteins 0.000 description 2
- 230000000981 bystander Effects 0.000 description 2
- 230000020411 cell activation Effects 0.000 description 2
- 230000005754 cellular signaling Effects 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- BFPSDSIWYFKGBC-UHFFFAOYSA-N chlorotrianisene Chemical compound C1=CC(OC)=CC=C1C(Cl)=C(C=1C=CC(OC)=CC=1)C1=CC=C(OC)C=C1 BFPSDSIWYFKGBC-UHFFFAOYSA-N 0.000 description 2
- 230000004940 costimulation Effects 0.000 description 2
- 238000012258 culturing Methods 0.000 description 2
- 108010086096 desmuslin Proteins 0.000 description 2
- 238000013401 experimental design Methods 0.000 description 2
- 108010002182 glycyl-phenylalanyl-lysyl-glutaminyl-seryl-seryl-lysyl-alanyl-leucine Proteins 0.000 description 2
- 108010014242 gp100(17-25) peptide Proteins 0.000 description 2
- 108010072094 gp100(280-288) melanoma antigen peptide Proteins 0.000 description 2
- 101150077981 groEL gene Proteins 0.000 description 2
- 108700025184 hepatitis B virus X Proteins 0.000 description 2
- 206010073071 hepatocellular carcinoma Diseases 0.000 description 2
- 231100000844 hepatocellular carcinoma Toxicity 0.000 description 2
- 102000048778 human Enah Human genes 0.000 description 2
- 108091005020 human MAGE-A1 protein (278-286) Proteins 0.000 description 2
- 102000028650 human MAGE-A1 protein (278-286) Human genes 0.000 description 2
- 108010007811 human immunodeficiency virus p17 gag peptide Proteins 0.000 description 2
- 210000002865 immune cell Anatomy 0.000 description 2
- 210000000987 immune system Anatomy 0.000 description 2
- 229960003444 immunosuppressant agent Drugs 0.000 description 2
- 230000001861 immunosuppressant effect Effects 0.000 description 2
- 239000003018 immunosuppressive agent Substances 0.000 description 2
- 229960000598 infliximab Drugs 0.000 description 2
- 208000037798 influenza B Diseases 0.000 description 2
- 108010061181 influenza matrix peptide (58-66) Proteins 0.000 description 2
- 229940125396 insulin Drugs 0.000 description 2
- 229960003130 interferon gamma Drugs 0.000 description 2
- 230000002147 killing effect Effects 0.000 description 2
- 108010003486 leucyl-leucyl-phenylalanyl-glycyl-tyrosyl-prolyl-valyl-tyrosyl-valine Proteins 0.000 description 2
- 210000004698 lymphocyte Anatomy 0.000 description 2
- 230000036210 malignancy Effects 0.000 description 2
- 238000010172 mouse model Methods 0.000 description 2
- 210000003205 muscle Anatomy 0.000 description 2
- IWPRJTUCJJRGSK-GOTSBHOMSA-N n-[(2s)-3-methyl-1-oxo-1-[[(2s)-1-oxo-3-phenylpropan-2-yl]amino]butan-2-yl]naphthalene-2-carboxamide Chemical compound C([C@H](NC(=O)[C@@H](NC(=O)C=1C=C2C=CC=CC2=CC=1)C(C)C)C=O)C1=CC=CC=C1 IWPRJTUCJJRGSK-GOTSBHOMSA-N 0.000 description 2
- 239000013642 negative control Substances 0.000 description 2
- 231100001083 no cytotoxicity Toxicity 0.000 description 2
- 239000004306 orthophenyl phenol Substances 0.000 description 2
- 244000045947 parasite Species 0.000 description 2
- 230000001575 pathological effect Effects 0.000 description 2
- 108010069653 peptide E (adrenal medulla) Proteins 0.000 description 2
- 230000001737 promoting effect Effects 0.000 description 2
- NGVDGCNFYWLIFO-UHFFFAOYSA-N pyridoxal 5'-phosphate Chemical compound CC1=NC=C(COP(O)(O)=O)C(C=O)=C1O NGVDGCNFYWLIFO-UHFFFAOYSA-N 0.000 description 2
- 102220095229 rs876658895 Human genes 0.000 description 2
- 210000004988 splenocyte Anatomy 0.000 description 2
- 238000010186 staining Methods 0.000 description 2
- 210000005050 synemin Anatomy 0.000 description 2
- 108010044720 telomerase reverse transcriptase (540-548) Proteins 0.000 description 2
- 238000002560 therapeutic procedure Methods 0.000 description 2
- 101710132362 truncated interferon antagonist D8 Proteins 0.000 description 2
- 108010059061 tyrosyl-leucyl-asparagyl-lysyl-isoleucyl-glutaminyl-asparagyl-seryl-leucine Proteins 0.000 description 2
- KIUKXJAPPMFGSW-DNGZLQJQSA-N (2S,3S,4S,5R,6R)-6-[(2S,3R,4R,5S,6R)-3-Acetamido-2-[(2S,3S,4R,5R,6R)-6-[(2R,3R,4R,5S,6R)-3-acetamido-2,5-dihydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-2-carboxy-4,5-dihydroxyoxan-3-yl]oxy-5-hydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-3,4,5-trihydroxyoxane-2-carboxylic acid Chemical compound CC(=O)N[C@H]1[C@H](O)O[C@H](CO)[C@@H](O)[C@@H]1O[C@H]1[C@H](O)[C@@H](O)[C@H](O[C@H]2[C@@H]([C@@H](O[C@H]3[C@@H]([C@@H](O)[C@H](O)[C@H](O3)C(O)=O)O)[C@H](O)[C@@H](CO)O2)NC(C)=O)[C@@H](C(O)=O)O1 KIUKXJAPPMFGSW-DNGZLQJQSA-N 0.000 description 1
- 102100024341 10 kDa heat shock protein, mitochondrial Human genes 0.000 description 1
- SGTNSNPWRIOYBX-UHFFFAOYSA-N 2-(3,4-dimethoxyphenyl)-5-{[2-(3,4-dimethoxyphenyl)ethyl](methyl)amino}-2-(propan-2-yl)pentanenitrile Chemical compound C1=C(OC)C(OC)=CC=C1CCN(C)CCCC(C#N)(C(C)C)C1=CC=C(OC)C(OC)=C1 SGTNSNPWRIOYBX-UHFFFAOYSA-N 0.000 description 1
- 241000251468 Actinopterygii Species 0.000 description 1
- 102100038910 Alpha-enolase Human genes 0.000 description 1
- 241000203069 Archaea Species 0.000 description 1
- 241000269837 Artemisia dubia Species 0.000 description 1
- 235000003261 Artemisia vulgaris Nutrition 0.000 description 1
- 101150013616 BHRF1 gene Proteins 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- 241000606124 Bacteroides fragilis Species 0.000 description 1
- 206010060999 Benign neoplasm Diseases 0.000 description 1
- 241000283725 Bos Species 0.000 description 1
- 241000283690 Bos taurus Species 0.000 description 1
- 238000011357 CAR T-cell therapy Methods 0.000 description 1
- 101710178629 ESAT-6-like protein Proteins 0.000 description 1
- 108010054218 Factor VIII Proteins 0.000 description 1
- 102000001690 Factor VIII Human genes 0.000 description 1
- 241000233866 Fungi Species 0.000 description 1
- OYRVWOGRRQDEQH-MLVLNPCWSA-N Gln-Tyr-Ile-Lys-Ala-Asn-Ser-Lys-Phe-Ile-Gly-Ile-Thr-Glu-Leu Chemical compound C([C@@H](C(=O)N[C@@H](C(C)CC)C(=O)NCC(=O)N[C@@H](C(C)CC)C(=O)N[C@@H](C(C)O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(O)=O)NC(=O)[C@H](CCCCN)NC(=O)[C@H](CO)NC(=O)[C@H](CC(N)=O)NC(=O)[C@H](C)NC(=O)[C@H](CCCCN)NC(=O)[C@@H](NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@@H](N)CCC(N)=O)C(C)CC)C1=CC=CC=C1 OYRVWOGRRQDEQH-MLVLNPCWSA-N 0.000 description 1
- 102100033945 Glycine receptor subunit alpha-1 Human genes 0.000 description 1
- BWMFRQKICHXLSH-ZZZNUFMVSA-N Grayanotoxin-III Natural products O[C@H]1[C@@H]2[C@@](O)(C)C[C@]31[C@H]([C@](O)(C)[C@H]1[C@](O)([C@H](O)C3)C(C)(C)[C@@H](O)C1)CC2 BWMFRQKICHXLSH-ZZZNUFMVSA-N 0.000 description 1
- 102000008949 Histocompatibility Antigens Class I Human genes 0.000 description 1
- 108010088652 Histocompatibility Antigens Class I Proteins 0.000 description 1
- 101000996297 Homo sapiens Glycine receptor subunit alpha-1 Proteins 0.000 description 1
- 101000808590 Homo sapiens Probable ubiquitin carboxyl-terminal hydrolase FAF-Y Proteins 0.000 description 1
- 101000738335 Homo sapiens T-cell surface glycoprotein CD3 zeta chain Proteins 0.000 description 1
- 101100122503 Human herpesvirus 6A (strain Uganda-1102) gN gene Proteins 0.000 description 1
- 206010062016 Immunosuppression Diseases 0.000 description 1
- 108700042652 LMP-2 Proteins 0.000 description 1
- 241000270322 Lepidosauria Species 0.000 description 1
- 241000209082 Lolium Species 0.000 description 1
- 108060001084 Luciferase Proteins 0.000 description 1
- 239000005089 Luciferase Substances 0.000 description 1
- 102000018697 Membrane Proteins Human genes 0.000 description 1
- 108010052285 Membrane Proteins Proteins 0.000 description 1
- 101150076359 Mhc gene Proteins 0.000 description 1
- 108010006519 Molecular Chaperones Proteins 0.000 description 1
- 241001529936 Murinae Species 0.000 description 1
- 241000699666 Mus <mouse, genus> Species 0.000 description 1
- 101100154912 Mus musculus Tyrp1 gene Proteins 0.000 description 1
- 108010058846 Ovalbumin Proteins 0.000 description 1
- 108010055531 P30 tetanus toxin peptide Proteins 0.000 description 1
- 101710125824 Pertussis toxin subunit 2 Proteins 0.000 description 1
- 108010022181 Phosphopyruvate Hydratase Proteins 0.000 description 1
- 108010064851 Plant Proteins Proteins 0.000 description 1
- 102100038600 Probable ubiquitin carboxyl-terminal hydrolase FAF-Y Human genes 0.000 description 1
- 102000004245 Proteasome Endopeptidase Complex Human genes 0.000 description 1
- 108090000708 Proteasome Endopeptidase Complex Proteins 0.000 description 1
- 108700039882 Protein Glutamine gamma Glutamyltransferase 2 Proteins 0.000 description 1
- 102100038095 Protein-glutamine gamma-glutamyltransferase 2 Human genes 0.000 description 1
- 241000713877 Simian sarcoma-associated virus Species 0.000 description 1
- 102100037906 T-cell surface glycoprotein CD3 zeta chain Human genes 0.000 description 1
- 101150104684 UL44 gene Proteins 0.000 description 1
- 241000251539 Vertebrata <Metazoa> Species 0.000 description 1
- 102100035071 Vimentin Human genes 0.000 description 1
- 108010065472 Vimentin Proteins 0.000 description 1
- 241000700605 Viruses Species 0.000 description 1
- 241000219094 Vitaceae Species 0.000 description 1
- 241000342876 [Clostridium] asparagiforme Species 0.000 description 1
- 210000005006 adaptive immune system Anatomy 0.000 description 1
- 230000007815 allergy Effects 0.000 description 1
- PLOPBXQQPZYQFA-AXPWDRQUSA-N amlintide Chemical compound C([C@@H](C(=O)NCC(=O)N[C@@H](C)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](C(C)C)C(=O)NCC(=O)N[C@@H](CO)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(N)=O)NC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CO)NC(=O)[C@H](CO)NC(=O)[C@H](CC=1NC=NC=1)NC(=O)[C@@H](NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CC=1C=CC=CC=1)NC(=O)[C@H](CC(N)=O)NC(=O)[C@H](C)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@@H](NC(=O)[C@H](C)NC(=O)[C@H]1NC(=O)[C@H]([C@@H](C)O)NC(=O)[C@H](C)NC(=O)[C@H]([C@@H](C)O)NC(=O)[C@H](CC(N)=O)NC(=O)[C@@H](NC(=O)[C@@H](N)CCCCN)CSSC1)[C@@H](C)O)C(C)C)C1=CC=CC=C1 PLOPBXQQPZYQFA-AXPWDRQUSA-N 0.000 description 1
- 230000030741 antigen processing and presentation Effects 0.000 description 1
- 230000000890 antigenic effect Effects 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 230000000599 auto-anti-genic effect Effects 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 229960003008 blinatumomab Drugs 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 230000000139 costimulatory effect Effects 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 230000006240 deamidation Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000010494 dissociation reaction Methods 0.000 description 1
- 230000005593 dissociations Effects 0.000 description 1
- 230000008029 eradication Effects 0.000 description 1
- 241001233957 eudicotyledons Species 0.000 description 1
- 229960000301 factor viii Drugs 0.000 description 1
- 235000019688 fish Nutrition 0.000 description 1
- 238000000684 flow cytometry Methods 0.000 description 1
- 230000004907 flux Effects 0.000 description 1
- 239000012634 fragment Substances 0.000 description 1
- 101150029683 gB gene Proteins 0.000 description 1
- 101150002378 gC gene Proteins 0.000 description 1
- 235000021312 gluten Nutrition 0.000 description 1
- 235000021021 grapes Nutrition 0.000 description 1
- 239000013574 grass pollen allergen Substances 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 229920002674 hyaluronan Polymers 0.000 description 1
- 229960003160 hyaluronic acid Drugs 0.000 description 1
- 238000011493 immune profiling Methods 0.000 description 1
- 230000001506 immunosuppresive effect Effects 0.000 description 1
- 238000009169 immunotherapy Methods 0.000 description 1
- 230000008676 import Effects 0.000 description 1
- 230000001965 increasing effect Effects 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 230000003834 intracellular effect Effects 0.000 description 1
- 235000021374 legumes Nutrition 0.000 description 1
- 108020001756 ligand binding domains Proteins 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 238000002483 medication Methods 0.000 description 1
- 230000003278 mimic effect Effects 0.000 description 1
- 230000004899 motility Effects 0.000 description 1
- 210000004498 neuroglial cell Anatomy 0.000 description 1
- 229940092253 ovalbumin Drugs 0.000 description 1
- 230000001717 pathogenic effect Effects 0.000 description 1
- 108010091748 peptide A Proteins 0.000 description 1
- 230000010412 perfusion Effects 0.000 description 1
- 235000021118 plant-derived protein Nutrition 0.000 description 1
- 239000004172 quinoline yellow Substances 0.000 description 1
- 230000009257 reactivity Effects 0.000 description 1
- 238000012552 review Methods 0.000 description 1
- 208000002491 severe combined immunodeficiency Diseases 0.000 description 1
- 210000003491 skin Anatomy 0.000 description 1
- 239000004402 sodium ethyl p-hydroxybenzoate Substances 0.000 description 1
- 230000009870 specific binding Effects 0.000 description 1
- 238000010561 standard procedure Methods 0.000 description 1
- 230000008685 targeting Effects 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 238000011282 treatment Methods 0.000 description 1
- 210000004881 tumor cell Anatomy 0.000 description 1
- 238000012795 verification Methods 0.000 description 1
- 210000005048 vimentin Anatomy 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/0005—Vertebrate antigens
- A61K39/001—Preparations to induce tolerance to non-self, e.g. prior to transplantation
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/46—Cellular immunotherapy
- A61K39/461—Cellular immunotherapy characterised by the cell type used
- A61K39/4611—T-cells, e.g. tumor infiltrating lymphocytes [TIL], lymphokine-activated killer cells [LAK] or regulatory T cells [Treg]
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/46—Cellular immunotherapy
- A61K39/462—Cellular immunotherapy characterized by the effect or the function of the cells
- A61K39/4621—Cellular immunotherapy characterized by the effect or the function of the cells immunosuppressive or immunotolerising
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/46—Cellular immunotherapy
- A61K39/463—Cellular immunotherapy characterised by recombinant expression
- A61K39/4631—Chimeric Antigen Receptors [CAR]
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/46—Cellular immunotherapy
- A61K39/463—Cellular immunotherapy characterised by recombinant expression
- A61K39/4632—T-cell receptors [TCR]; antibody T-cell receptor constructs
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/46—Cellular immunotherapy
- A61K39/464—Cellular immunotherapy characterised by the antigen targeted or presented
- A61K39/4643—Vertebrate antigens
- A61K39/46433—Antigens related to auto-immune diseases; Preparations to induce self-tolerance
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/46—Cellular immunotherapy
- A61K39/464—Cellular immunotherapy characterised by the antigen targeted or presented
- A61K39/4643—Vertebrate antigens
- A61K39/46434—Antigens related to induction of tolerance to non-self
-
- 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
- C07K14/70503—Immunoglobulin superfamily
- C07K14/7051—T-cell receptor (TcR)-CD3 complex
-
- 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
- C07K14/70503—Immunoglobulin superfamily
- C07K14/70539—MHC-molecules, e.g. HLA-molecules
-
- 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
- C07K14/70596—Molecules with a "CD"-designation not provided for elsewhere
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
- C12N15/102—Mutagenizing nucleic acids
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2319/00—Fusion polypeptide
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2319/00—Fusion polypeptide
- C07K2319/01—Fusion polypeptide containing a localisation/targetting motif
- C07K2319/02—Fusion polypeptide containing a localisation/targetting motif containing a signal sequence
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2319/00—Fusion polypeptide
- C07K2319/01—Fusion polypeptide containing a localisation/targetting motif
- C07K2319/03—Fusion polypeptide containing a localisation/targetting motif containing a transmembrane segment
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2319/00—Fusion polypeptide
- C07K2319/33—Fusion polypeptide fusions for targeting to specific cell types, e.g. tissue specific targeting, targeting of a bacterial subspecies
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2319/00—Fusion polypeptide
- C07K2319/70—Fusion polypeptide containing domain for protein-protein interaction
- C07K2319/73—Fusion polypeptide containing domain for protein-protein interaction containing coiled-coiled motif (leucine zippers)
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2319/00—Fusion polypeptide
- C07K2319/80—Fusion polypeptide containing a DNA binding domain, e.g. Lacl or Tet-repressor
- C07K2319/81—Fusion polypeptide containing a DNA binding domain, e.g. Lacl or Tet-repressor containing a Zn-finger domain for DNA binding
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Definitions
- TECHNICAL FIELD [0003] The technology described herein relates to chimeric antigen ligand (CAL) technology, e.g., for treating of autoimmune and/or T-cell mediated conditions.
- CAL chimeric antigen ligand
- BACKGROUND [0004]
- Current treatments for preventing autoimmunity and transplant rejection involve stringent immunosuppression, which can lead to severe, unwanted side effects including extreme susceptibility to infection, malignant and benign neoplasms, multiple organ and systems failure.
- T cells recognize their target cells by using receptors on their cell surface which are called T Cell Receptors (TCRs).
- TCRs have a recognition portion and a signaling portion. When the recognition portion binds to the natural complexes formed in the body in the presence of a diseased cell, the signaling portion is activated, which leads to the T cell engaging in killing activity or recruiting other immune cells to destroy the diseased cell.
- CAR-T cell therapy is known and seeks to help T cells recognize autoreactive and alloreactive T cells. This is accomplished by genetically altering a T cell so that it expresses a chimeric antigen receptor (CAR).
- CAR is an altered TCR, in which the natural recognition portion is removed and replaced with a synthetic recognition portion that is designed to more effectively recognize the autoreactive and alloreactive T cells by very specifically detecting the presence of a molecule unique to the autoreactive and alloreactive T cells.
- CAL chimeric antigen ligands
- the CALs described herein comprise a TCR recognition domain and a biomolecular interaction domain. The TCR recognition permits the CAL to bind to autoreactive and/or alloreactive T cells in an antigen specific manner.
- the biomolecular interaction domain permits an immune killer cell (e.g., a NK cell, a T cell, or a dendritic cell) to bind to the CAL, thereby promoting killing of the autoreactive and/or alloreactive T cell by the immune killer cell.
- the TCR recognition domain of a CAL binds specifically to a TCR, e.g., a TCR expressed on the surface of an autoreactive and/or alloreactive T cell.
- Exemplary but non-limiting TCR recognition domains include peptide-MHC complexes, e.g., in monomeric, oligomeric, or multimeric form.
- the TCR recognition domain can comprise natural or synthetic sequences.
- the biomolecular interaction domain of the CAL permits specific binding of the CAL with a second biomolecule, e.g., a receptor on the immune killer cell.
- a second biomolecule e.g., a receptor on the immune killer cell.
- the biomolecular interaction domain of the CAL is recognized by an endogenous receptor on the immune killer cell.
- the biomolecular interaction domain of the CAL is recognized by an engineered receptor on the immune killer cell.
- biomolecular interaction domains include FITC (which can be recognized by a FITC CAR-T cell system), a leucine zipper domain, a zinc finger domain, PSD95-Dlg1-zo-1 (PDZ) domains, a streptavidin domain and a streptavidin binding protein (SBP) domain, a gibberellin insensitive (GIA) and/or a gibberellin insensitive dwarf1 (GID1), a PYL domain and an ABI domain, a chemically-induced pair of interaction domains as described elsewhere herein, a Snap-tag, a Halo tag, a T14-3-3- cdeltaC and/or a C-Terminal peptide of PMA2 (CT52), a (R)-Phycoerythrin (R-PE/ PE) and/or a R-PE/PE binding protein a Fab domain, and/or an anti-CD3 domain.
- FITC which can be recognized by a FITC CAR-T cell system
- CALs described herein can be used in combination with CAR-T systems.
- Exemplary but non-limiting CAR-T systems suitable for use with the methods and compositions described herein include SUPRA CAR and SPLIT CAR.
- CAR-T systems are discussed in more detail elsewhere herein and are known in the art.
- a composition comprising: a) a TCR recognition domain; and one or both of: b) an intracellular signaling domain; and c) a biomolecular interaction domain (e.g., a first-type biomolecular interaction domain).
- composition comprising: a first polypeptide comprising at least a portion of a TCR recognition domain and a first-type biomolecular interaction domain; and a signaling polypeptide comprising a second-type biomolecular interaction domain and an intracellular signaling domain; wherein the first-type and second-type biomolecular interaction domains bind specifically to each other.
- composition comprising: a first polypeptide comprising at least a portion of a TCR recognition domain and a first-type biomolecular interaction domain; and a recognition polypeptide comprising a second recognition domain and a third-type biomolecular interaction domain; wherein the first-type and third-type biomolecular interaction domains bind specifically to each other.
- composition comprising: a first polypeptide comprising at least a portion of a TCR recognition domain and a first-type biomolecular interaction domain; and a signaling polypeptide comprising a second-type biomolecular interaction domain and an intracellular signaling domain; and a recognition polypeptide comprising a second recognition domain and a third-type biomolecular interaction domain; wherein the second-type and third-type biomolecular interaction domains compete for binding to the first-type biomolecular interaction domain.
- the third-type biomolecular interaction domain and first- type biomolecular interaction domain have a higher affinity for each other than the second- type biomolecular interaction domain and first-type biomolecular interaction domain.
- described herein is a composition comprising: a first polypeptide comprising at least a portion of a TCR recognition domain and a first-type biomolecular interaction domain; a signaling polypeptide comprising a second-type biomolecular interaction domain, a fourth-type biomolecular interaction domain, and an intracellular signaling domain; and a recognition polypeptide comprising a second recognition domain and a fifth-type biomolecular interaction domain; wherein the first-type biomolecular interaction domain and the second-type biomolecular interaction domain bind specifically to each other; and wherein the fourth-type biomolecular interaction domain and the fifth-type biomolecular interaction domain bind specifically to each other.
- the fourth-type biomolecular interaction domain and fifth-type biomolecular interaction domain have a weaker affinity than the second-type biomolecular interaction domain and first-type protein interaction domain.
- the first polypeptide further comprises a sixth-type biomolecular interaction domain and the recognition polypeptide further comprises a seventh-type biomolecular interaction domain which bind specifically to each other. [0013] In some embodiments of any of the aspects, the first polypeptide comprises the entire TCR recognition domain.
- the TCR recognition domain comprises at least two separate polypeptide sequences
- the first polypeptide comprises at least one of the separate polypeptide sequences of the TCR recognition domain
- the first polypeptide is bound to or complexed with a second or further polypeptide sequences of the TCR recognition domain to form a TCR recognition domain.
- the TCR recognition domain comprises a non-polypeptide component.
- the second recognition domain is specific for a target that is not recognized by the TCR recognition domain.
- the second recognition domain is specific for a target that is found on a healthy and/or non-target cell and not on a diseased and/or target cell.
- the TCR recognition domain comprises a MHC (Major Histocompatibility Complex); a MHC-peptide complex; featureless peptide MHC; or a MHC-peptide fusion.
- the peptide is a human or non-human peptide.
- the peptide is a Minor Histocompatibility Antigen (MiHA).
- the MHC is a monomer, dimer, trimer, tetramer, pentamer, dextramer or other oligomer form.
- the MHC-peptide complex is a monomer, dimer, trimer, tetramer, pentamer, dextramer or other oligomer form.
- the MHC-peptide fusion is a monomer, dimer, trimer, tetramer, pentamer, dextramer or other oligomer form.
- the MHC is a dimer, trimer, tetramer, pentamer, dextramer or other oligomer form.
- the MHC-peptide complex is a dimer, trimer, tetramer, pentamer, dextramer or other oligomer form.
- the MHC-peptide fusion is a dimer, trimer, tetramer, pentamer, dextramer or other oligomer form.
- the MHC is a MHC class I or a MHC class II.
- the TCR recognition domain comprises a CD1 domain or a CD1 domain-ligand complex or fusion.
- the CD1 is CD1d.
- the biomolecular interaction domains are found on an extracellular portion of the respective polypeptides.
- biomolecular interaction domain(s) is a BZip (RR) and/or a AZip (EE), or any binding pair of biomolecular interaction domains are collectively a BZip (RR) and a AZip (EE); c. wherein the biomolecular interaction domain(s) is a PSD95-Dlgl-zo-1 (PDZ) domain; d. wherein the biomolecular interaction domain(s) is a streptavidin and/or a streptavidin binding biomolecular (SBP) or any binding pair of biomolecular interaction domains are collectively a streptavidin and a streptavidin binding biomolecular (SBP); e.
- PDZ PSD95-Dlgl-zo-1
- biomolecular interaction domain(s) is a FKBP-binding domain of mTOR (FRB) and/or a FK506 binding biomolecular (FKBP) or any binding pair of biomolecular interaction domains are collectively a FKBP-binding domain of mTOR (FRB) and a FK506 binding biomolecular (FKBP); f.
- biomolecular interaction domain(s) is a cyclophilin-Fas fusion biomolecular (CyP-Fas) and/or a FK506 binding biomolecular (FKBP) or any binding pair of biomolecular interaction domains are collectively a cyclophilin-Fas fusion biomolecular (CyP-Fas) and a FK506 binding biomolecular (FKBP); g.
- biomolecular interaction domain(s) is a calcineurin A (CNA) and/or a FK506 binding biomolecular (FKBP) or any binding pair of biomolecular interaction domains are collectively a calcineurin A (CNA) and a FK506 binding biomolecular (FKBP); h. wherein the biomolecular interaction domain(s) is a gibberellin insensitive (GIA) and/or a gibberellin insensitive dwarf1 (GID1) or any binding pair of biomolecular interaction domains are collectively a gibberellin insensitive (GIA) and a gibberellin insensitive dwarf1 (GID1); i.
- biomolecular interaction domain(s) is a Snap-tag and/or a Halo tag, or any binding pair of biomolecular interaction domains are collectively a Snap-tag and a Halo tag; j. wherein the biomolecular interaction domain(s) is a T14-3-3-cdeltaC and/or a C- Terminal peptides of PMA2 (CT52), or any binding pair of biomolecular interaction domains are collectively a T14-3-3-cdeltaC and a C-Terminal peptides of PMA2 (CT52); k.
- biomolecular interaction domain(s) is a PYL and/or a ABI, or any binding pair of biomolecular interaction domains are collectively a PYL and a ABI; l. wherein the biomolecular interaction domain(s) is a nucleotide tag and/or a zinc finger domain, or any binding pair of biomolecular interaction domains are collectively a nucleotide tag and a zinc finger domain; m. wherein the biomolecular interaction domain(s) is a nucleotide tag, or any binding pair of biomolecular interaction domains are collectively a pair of nucleotide tags; n.
- biomolecular interaction domain(s) is a Fluorescein isothiocyanate (FITC) and/or a FITC binding biomolecular or any binding pair of protein interaction domains are collectively a FITC and a FITC binding protein; and/or o.
- the protein interaction domain(s) is a (R)-Phycoerythrin (R-PE/ PE) and/or a R-PE/PE binding protein or any binding pair of protein interaction domains are collectively a (R)-Phycoerythrin (R-PE/ PE) and a R-PE/PE binding protein.
- the nucleotide tag is a DNA tag or dsDNA tag.
- the intracellular signaling domain comprises or is a signaling domain from one or more proteins selected from the group consisting of: TCR ⁇ , FcR ⁇ , FcR ⁇ , CD3 ⁇ ; CD35; CD3 ⁇ ; CD3C; CD22; CD79a; CD79b; CD66d; CARD11; CD2; CD7; CD27; CD28; CD30; CD40; CD54 (ICAM); CD83; CD134 (OX40); CD137 (4-1BB); CD150 (SLAMF1); CD152 (CTLA4); CD223 (LAG3); CD270 (HVEM); CD273 (PD-L2); CD274 (PD- Ll); CD278 (ICOS); DAP10; LAT; KD2C SLP76; TRIM; and ZAP70.
- proteins selected from the group consisting of: TCR ⁇ , FcR ⁇ , FcR ⁇ , CD3 ⁇ ; CD35; CD3 ⁇ ; CD3C; CD22; CD79a; CD79b; CD66d; CARD11; CD
- the TCR recognition domain comprises a MHC allogeneic, autologous, or xenogeneic to the cell. In some embodiments of any of the aspects, the TCR recognition domain comprises a synthetic MHC.
- the TCR recognition domain comprises a MHC and a peptide, wherein the peptide is allogeneic, autologous, or xenogeneic to the cell. In some embodiments of any of the aspects, the TCR recognition domain comprises a MHC and a peptide, wherein the peptide is synthetic.
- the cell is a NK cell, dendritic cell, regulatory T cell, or effector T cell. In some embodiments of any of the aspects, the cell is engineered to express one of more of the polypeptide(s) of the composition. In some embodiments of any of the aspects, the cell is engineered to express the signaling polypeptide of the composition.
- the cell is further engineered to knockout or knockdown the native MHCI/II. In some embodiments of any of the aspects, the cell is further engineered to knockdown the native MHCI/II expressed on the cell surface.
- a composition comprising a TCR recognition domain and an intracellular signaling domain but not comprising a biomolecular interaction domain (e.g., a first-type biomolecular interaction domain).
- a composition comprising a TCR recognition domain and a biomolecular interaction domain (e.g., a first- type biomolecular interaction domain) and but not comprising an intracellular signaling domain.
- a biomolecular interaction domain e.g., a first- type biomolecular interaction domain
- an intracellular signaling domain e.g., a first- type biomolecular interaction domain
- described herein is a method of treating or preventing an autoimmune disease or condition; T cell mediated inflammation or immune response; malignant T cell condition; transplant rejection; or GvHD in a subject in need thereof, the method comprising administering to the subject a composition and/or cell of any of the preceding claims.
- the TCR recognition domain comprises a MHC allogeneic to the subject.
- the TCR recognition domain comprises a MHC autologous to the transplant cells. In some embodiments of any of the aspects, the TCR recognition domain comprises a MHC xenogeneic to the transplant cells. In some embodiments of any of the aspects, the TCR recognition domain comprises a MHC and a peptide, wherein the peptide is allogeneic to the subject. In some embodiments of any of the aspects, the TCR recognition domain comprises a MHC and a peptide, wherein the peptide is autologous to the transplant cells. In some embodiments of any of the aspects, the TCR recognition domain comprises a MHC and a peptide, wherein the peptide is autologous to the transplant cells.
- the transplant is any human or non-human cell, tissue, or organ. In some embodiments of any of the aspects, the transplant is an allogeneic hematopoietic stem cell or solid organ transplantation.
- the malignant T cell condition is T cell acute lymphoblastic leukemia or T cell lymphoblastic lymphoma.
- the autoimmune disease is type 1 diabetes, vitiligo, multiple sclerosis, alopecia, celiac disease, pemphigus, rheumatoid arthritis, or scleroderma.
- the autoimmune disease is thyroiditis, type 1 diabetes mellitus, Hashimoto's thyroiditis, Graves' disease, celiac disease, multiple sclerosis, Guillain-Barre syndrome, Addison's disease, and Raynaud's phenomenon, Goodpasture's disease, arthritis (rheumatoid arthritis such as acute arthritis, chronic rheumatoid arthritis, gout or gouty arthritis, acute gouty arthritis, acute immunological arthritis, chronic inflammatory arthritis, degenerative arthritis, type II collagen-induced arthritis, infectious arthritis, Lyme arthritis (e.g., post treatment Lyme disease syndrome), proliferative arthritis, psoriatic arthritis, Still's disease, vertebral arthritis, and juvenile-onset rheumatoid arthritis, arthritis chronica progrediente, arthritis deformans, polyarthritis chronica primaria, reactive arthritis, and ankylosing spondylitis), palindromic arthritis, inflammatory hyperprolifera, arthritis chronica progredient
- the T cell mediated immune response is an anti-drug specific response to a biologic, cell therapy, and/or gene therapy.
- the biologic, cell-therapy, or gene therapy is an adeno-associated virus (AAV) gene therapy, a genome editing agent, or enzyme replacement therapy.
- AAV adeno-associated virus
- the disease is type 1 diabetes and the TCR recognition domain comprises sequences with at least 80% or at least 95% sequence identity to: one or more of SEQ ID NOs: 8-17; HLA-A*0201 and at least one of SEQ ID NOs: 2013-2016 and 2031-2033; or HLA-A*02:01 and at least one of SEQ ID NOs: 20128-2129.
- the disease is vitiligo and the TCR recognition domain comprises sequences with at least 80% or at least 95% sequence identity to: SEQ ID NO: 18, 19, and one of 20-22; or comprises HLA-A*0201 and SEQ ID NO: 2018; or HLA-A*0301 and SEQ ID NO: 2019; or comprises HLA-A*2402 and SEQ ID NO: 2020; or HLA-A*0101 and SEQ ID NO: 2021.
- the method is a method of treating and/or preventing GvHD and the TCR recognition domain comprises sequences with at least 80% or at least 95% sequence identity to: HLA-A*0101 and at least one of SEQ ID NOs: 2034-2037; or HLA-B*0702 and SEQ ID NO: 2038; or HLA-B*0801 and SEQ ID NO: 2039.
- the disease is type 1 diabetes and the TCR recognition domain comprises one or more of SEQ ID NOs: 8-17; comprises HLA-A*0201 and at least one of SEQ ID NOs: 2013-2016 and 2031-2033; or comprises HLA-A*02:01 and at least one of SEQ ID NOs: 20128-2129.
- the disease is vitiligo and the TCR recognition domain comprises SEQ ID NO: 18, 19, and one of 20-22; or comprises HLA- A*0201 and SEQ ID NO: 2018; or comprises HLA-A*0301 and SEQ ID NO: 2019; or comprises HLA-A*2402 and SEQ ID NO: 2020; or comprises HLA-A*0101 and SEQ ID NO: 2021.
- the method is a method of treating and/or preventing GvHD and the TCR recognition domain comprises HLA-A*0101 and at least one of SEQ ID NOs: 2034-2037; or comprises HLA-B*0702 and SEQ ID NO: 2038; or comprises HLA-B*0801 and SEQ ID NO: 2039.
- Fig.1 demonstrates that pMHC tetramer + FITC (adaptor) binds to the target cells (OTi) in a dose dependent fashion or manner.
- I-H2Kb MHC class I tetramer.
- I-Ab- Control tetramer.
- Figure discloses SEQ ID NOS 2752-2753, 2750 and 2754, respectively, in order of appearance.
- Fig.2 depicts expression of activation marker CD69 on OTi cells at different time points (24, 48, 72 hrs.). Binding of Jurkat+ pMHC to OTi cells does not change Jurkat live count (right).
- Figure discloses SEQ ID NOS 2750, 2750 and 2754, respectively, in order of appearance.
- Fig.3 depicts the expression of CD69 on Jurkat CAR, e.g., CAL T cells in different time points (24, 48, 72 hrs.).
- Figure discloses SEQ ID NOS 2750 and 2755, respectively, in order of appearance.
- Fig.4 depicts the cytotoxicity of Primary human CD8 CAR, e.g., CAL T cells with different concentrations of tetramers (left).
- Minimal activation of target cells CD69 on CAR, e.g., CAL T cells and target cells
- I-Ab Ctrl/ H2Kb: MHC-I.
- Figure discloses SEQ ID NOS 2750, 2754 and 2750, respectively, in order of appearance.
- Fig.5 demonstrates that the cytotoxicity of human CD8 CAR , e.g., CAL T is highly specific and was not seen with ctrl tetramer (left).
- Fig.6A depicts a schematic of SUPRA CAR design applied to provide Universal CAL. This design separates the cell targeting molecule module from the killer cell.
- Fig.6B depicts that SUPRA CARs can be designed with CD3 ⁇ domain uncoupled from the costimulatory domains to provide Universal CAL as described herein.
- Figs.7A-7D depict key features of the SUPRA CAR systems that can be applied to Universal CAL.
- Fig.7A demonstrates that zipCAR activation is tunable through modulation of zipFv concentration, zipper affinity, scFv affinity, and zipCAR expression level in human primary CD4 T cells, as demonstrated by IFN-g production.
- Fig.7B demonstrates that SUPRA CAR system, as applied to Universal CAL,can perform combination antigen detection to form AND gate logic in CD4 T cells.
- Fig.7C demonstrates that xenograft animal tumor model shows tumor eradication (as demonstrated by luciferase photon flux given by the tumor cells) by the SUPRA CAR T cells.
- Fig.7D demonstrates that SUPRA CARs as applied to Universal CAL can be used to control different cell types, such as CD4 and CD8 T cells, against two different antigens.
- CD69 expression (a T cell activation marker) is quantified with flow cytometry for CD4 and CD8 T cells.
- Fig.8 depicts the timeline of double Hu-PBMC-HSCT-skin graft mouse model generation.
- Fig.9 depicts a summary of double hu-PBMC-HSCT-skin graft mouse model generation.
- Fig.10 depicts key features of pMHC multimer + CAR, e.g., CAL T cell system.
- Fig.11 depicts a table of experimental design.
- Figure discloses SEQ ID NOS 2750, 2756 and 2754, respectively, in order of appearance.
- Fig.12 demonstrates verification of FITC-conjugated tetramer mediated activation.
- Figure discloses SEQ ID NOS 2750 and 2751, respectively, in order of appearance.
- Fig.13 depicts a time course of FITC-conjugated tetramer mediated activation.
- Figure discloses SEQ ID NOS 2750, 2751, 2750, 2751, 2750, 2751, 2750 and 2751, respectively, in order of appearance.
- Fig.14 depicts a graph of Jurkat cell counts.
- Figure discloses SEQ ID NOS 2750 and 2754, respectively, in order of appearance.
- Fig.15 depicts tetramer staining.
- Figure discloses SEQ ID NOS 2750, 2754, 2751, 2756, 2754, 2750 and 2754, from left to right and top to bottom.
- Fig.16 depicts heatmaps of indicated staining levels.
- Figure discloses SEQ ID NOS 2751, 2757, 2754, 2750, 2754, 2751, 2754, 2757, 2750, 2754, 2751, 2754, 2757, 2750 and 2754, respectively, in order of appearance.
- Fig.17 depicts a table of experimental design.
- Figs.18-20 depict graphs of cytotoxicity levels.
- Fig.18 discloses SEQ ID NOS 2754, 2750, 2754 and 2750 from left to right.
- Fig.19 discloses SEQ ID NOS 2754 and 2750, respectively, in order of appearance.
- Fig.20 discloses SEQ ID NOS 2750, 2754, 2750 and 2754 from left to right.
- Fig.21 depicts the levels of CD69 on OTi CD8 T cells.
- Figure discloses SEQ ID NOS 2750, 2754, 2750 and 2754 from left to right.
- Fig.22 depicts schematics of two embodiments of the technology described herein.
- Fig.23 depicts a schematic of the FU-CAL embodiments of the technology described herein.
- Fig.24 depicts a schematic of the CAL-BITE embodiments of the technology described herein.
- the left panel depicts blinatumomab, which is described in more detail in Weiner et al. The Molecular Basis of Cancer 2015683-694.e3.
- Fig.25 depicts a schematic of the CAL technology disarming autoreactive T cells.
- Figs.26A-26B depict schematics of T cells design.
- MHC can be mouse or human.
- the MiHA can be ovalbumin (against OTi or OTii) or disparate antigens between donor and recipient.
- Fig.27 depicts a graph demonstrating that pMHC tetramer + FITC (adaptor) binds to the target cells (OTi) in a dose dependent fashion, while OTii specific tetramer does not.
- Fig.28A depicts expression of CD69 on OTi cells at different time points (24, 48, 72 hrs.).
- Fig.28B is a graph demonstrating that binding of Jurkat+ pMHC to OTi cells does not change Jurkat live count.
- Fig.29 depicts a graph demonstrating that cytotoxicity of pMHC-CAR against 1E6 T cell clone exhibits a dose dependent association.
- Fig.32 depicts a graph demonstrating cytotoxicity of Primary CD8 CAR T cells against OTi TCR T cells with different concentration of tetramer.
- DETAILED DESCRIPTION [0059] Aspects of the invention described herein relate to chimeric antigen ligands.
- chimeric antigen ligand or “CAL” refers to an artificially constructed molecule comprising a TCR recognition domain (e.g. an polypeptide comprising at least one MHC sequence as described herein) and at least one biomolecular interaction domain.
- the TCR recognition domain is selected to bind to specific populations of T cells that it is desirable to target and/or destroy, e.g., for therapeutic purposes in T-cell mediated diseases.
- the population of targeted T cells is a population of polyclonal pathogenic T cells.
- the biomolecular interaction domain is selected to bind to a second cell, e.g., a NK cell, thereby colocalizing the targeted T cell and the second cell and promoting or increasing the inhibition and/or destruction of the targeted T cell.
- the CAL is selected to have high affinity or avidity for the TCR, e.g., the TCR variable domain.
- the CALs can be used herein with endogenous cells, e.g., in some embodiments, no engineered cells are administered to the subject. In other embodiments, the CALs can be used with engineered cells, e.g., engineered NK cells.
- the engineered cells can comprise one or more CARs, e.g, a CAR comprising an extracellular domain with a biomolecular interaction domain that specifically binds with the biomolecular interaction domain of the CAL.
- CARs chimeric antigen receptors
- the two separate polypeptides that make up a complete CAR are able to interact and form the complete CAR by way of protein interaction domains. This permits flexible, modular CAR-T therapy which is capable of complex logic computation, providing a more precise and effective approach to immunotherapy.
- a CAL and/or chimeric antigen receptor having multiple components, and/or a cell or composition comprising a multi-component CAL and/or CAR.
- Multi-component CALs/CARs are also referred to herein variously as SMART CAL/CAR or SUPRA.
- traditional “chimeric antigen receptor” or “CAR” refers to an artificially constructed hybrid polypeptide comprising an antigen-binding domain (e.g. an antigen-binding portion of an antibody (e.g. a scFV)) linked to a cell signaling and/or cell activation domain.
- the cell-signaling domain can be a T-cell signaling domain.
- the cell activation domain can be a T-cell activation domain.
- CARs have the ability to redirect the specificity and reactivity of T cells and other immune cells (e.g., NK cells) toward a selected target in a non-MHC-restricted manner, exploiting the antigen-binding properties of monoclonal antibodies.
- NK cells immune cells
- the non-MHC-restricted antigen recognition gives T-cells expressing CARs the ability to recognize an antigen independent of antigen processing, thus bypassing a major mechanism of tumor escape.
- CARs when expressed in T-cells, CARs advantageously do not dimerize with endogenous T-cell receptor (TCR) alpha and beta chains.
- TCR T-cell receptor
- the CAR's extracellular binding domain is composed of a single chain variable fragment (scFv) derived from fusing the variable heavy and light regions of a murine or humanized monoclonal antibody.
- scFvs may be used that are derived from Fabs (instead of from an antibody, e.g., obtained from Fab libraries), in various embodiments, this scFv is fused to a transmembrane domain and then to an intracellular signaling domain.
- First- generation CARs include those that solely provide CD3zeta signals upon antigen binding
- “Second- generation” CARs include those that provide both costimulation (e.g. CD28 or CD 137) and activation (CD3Q).
- “Third-generation” CARs include those that provide multiple costimulation (e.g. CD28 and CD 137) and activation (CO3Q).
- the CAR is selected to have high affinity or avidity for the antigen. Further discussion of CARs can be found, e.g., in Maus et al. Blood 2014123:2624-35; Reardon et al. Neuro-Oncology 201416:1441-1458; Hoyos et al.
- multi-component CAL refers to a CAL comprising at least two separate polypeptides, neither of which polypeptides is capable of both ligand recognition and signaling activation on its own.
- multi-component CAR refers to a CAR comprising at least two separate polypeptides, neither of which polypeptides is capable of both ligand recognition and signaling activation on its own.
- the at least two separate polypeptides each comprise a protein interaction domain that permits interaction, e.g., binding of the separate polypeptides.
- one of the at least two separate polypeptides is a transmembrane polypeptide having an intracellular T cell receptor (TCR) signaling domain and a second of the at least two separate polypeptides is an extracellular polypeptide having a ligand-binding domain.
- TCR T cell receptor
- a multi-component CAL and/or CAR can comprise two, three, four, five, six, seven, eight, nine, ten or more separate polypeptides.
- Various aspects provided herein provide a composition comprising multiple components of a multi-component CAL and/or CAR.
- a composition e.g., a single molecule, comprising a TCR recognition domain; and one or both of: (a) an intracellular signaling domain; and (b) a first-type protein interaction domain.
- compositions e.g., a single molecule comprising a TCR recognition domain; and a first-type biomolecular (e.g., protein) interaction domain.
- a multi-component CAL and/or CAR comprising a TCR recognition domain; and one or both of: (a) an intracellular signaling domain; and (b) a first-type protein interaction domain.
- the composition, e.g, single molecule, comprising a TCR recognition domain; and a first-type biomolecular (e.g., protein) interaction domain does not comprise an antibody, antibody domain, or antibody reagent.
- compositions comprising (a) a first polypeptide comprising a TCR recognition domain and a first-type protein interaction domain; and (b) a signaling polypeptide comprising a second-type protein interaction domain and an intracellular signaling domain; wherein the first-type and second-type protein interaction domains bind specifically to each other.
- a multi-component CAL and/or CAR comprising a) a first polypeptide comprising a TCR recognition domain and a first-type protein interaction domain; and (b) a signaling polypeptide comprising a second-type protein interaction domain and an intracellular signaling domain; wherein the first-type and second-type protein interaction domains bind specifically to each other.
- compositions comprising (a) a first polypeptide comprising a TCR recognition domain and a first-type protein interaction domain; and (b) a recognition polypeptide comprising a second recognition domain and a third-type protein interaction domain; wherein the first-type and third-type protein interaction domains bind specifically to each other.
- a multi-component CAL and/or CAR comprising (a) a first polypeptide comprising a TCR recognition domain and a first-type protein interaction domain; and (b) a recognition polypeptide comprising a second recognition domain and a third-type protein interaction domain; wherein the first-type and third-type protein interaction domains bind specifically to each other [0070]
- Another aspect of the embodiments is a composition comprising (a) a first polypeptide comprising a TCR recognition domain and a first-type protein interaction domain; and (b) a signaling polypeptide comprising a second-type protein interaction domain and an intracellular signaling domain; and (c) a recognition polypeptide comprising a second recognition domain and a third-type protein interaction domain; wherein the second-type and third-type protein interaction domains compete for binding to the first-type protein interaction domain.
- a multi- component CAL and/or CAR comprising (a) a first polypeptide comprising a TCR recognition domain and a first-type protein interaction domain; and (b) a signaling polypeptide comprising a second-type protein interaction domain and an intracellular signaling domain; and (c) a recognition polypeptide comprising a second recognition domain and a third-type protein interaction domain; wherein the second-type and third-type protein interaction domains compete for binding to the first- type protein interaction domain.
- the third-type protein interaction domain and first-type protein interaction domain have a higher affinity for each other than the second-type protein interaction domain and first-type protein interaction domain.
- Affinity can be measured by one skilled in the art using standard methods, for example, by measuring its equilibrium dissociation constant (K d ).
- Another aspect of the embodiments is a composition comprising (a) a first polypeptide comprising a TCR recognition domain and a first-type protein interaction domain; (b) a signaling polypeptide comprising a second-type protein interaction domain, a fourth-type protein interaction domain, and an intracellular signaling domain; and (c) a recognition polypeptide comprising a second recognition domain and a fifth-type protein interaction domain; wherein the first-type protein interaction domain and the second-type protein interaction domain bind specifically to each other; and wherein the fourth-type protein interaction domain and the fifth-type protein interaction domain bind specifically to each other.
- a multi-component CAL and/or CAR comprising (a) a first polypeptide comprising a TCR recognition domain and a first-type protein interaction domain; (b) a signaling polypeptide comprising a second-type protein interaction domain, a fourth- type protein interaction domain, and an intracellular signaling domain; and (c) a recognition polypeptide comprising a second recognition domain and a fifth-type protein interaction domain; wherein the first-type protein interaction domain and the second-type protein interaction domain bind specifically to each other; and wherein the fourth-type protein interaction domain and the fifth-type protein interaction domain bind specifically to each other.
- the fourth-type protein interaction domain and fifth-type protein interaction domain have a weaker affinity than the second-type protein interaction domain and first- type protein interaction domain. Affinity can be measured as described above.
- the first polypeptide further comprises a sixth-type protein interaction domain and the recognition polypeptide further comprises a seventh-type protein interaction domain which bind specifically to each other.
- the first polypeptide comprises the entire TCR recognition domain.
- the TCR recognition domain comprises at least two separate polypeptide sequences
- the first polypeptide comprises at least one of the separate polypeptide sequences of the TCR recognition domain
- the first polypeptide is bound to or complexed with a second or further polypeptide sequences of the TCR recognition domain to form a TCR recognition domain.
- a composition described herein can comprise multiple copies or instances of a TCR recognition domain(s), e.g. the TCR recognition domain can be a mulitmer, or oligomer.
- a composition described herein can comprise multiple copies or instances of a first polypeptide as described herein.
- the second recognition domain is specific for a target that is not recognized by the TCR recognition domain. In one embodiment, the second recognition domain is specific for a target that is found on a healthy and/or non-target cell and not on a diseased and/or target cell.
- TCR recognition domain refers to a domain or portion of a polypeptide that can target or bind specifically to a TCR, e.g., a TCR expressed on the surface of T cell.
- the TCR recognition domain can be a TCR variable region (TCR-VR) recognition domain, i.e.
- the TCR recognition domain sequence can be autologous, allogeneic, or xenogeneic to a given subject.
- the TCR recognition domain sequence is a wild-type protein or sequence.
- the TCR recognition domain sequence is a naturally-occurring variant, e.g., an allele of a wild-type protein or sequence.
- the TCR recognition domain sequence is modified relative to a wild-type protein, e.g. chemically modified.
- the TCR recognition domain sequence is a derivative and/or variant of a wild-type sequence.
- the TCR recognition domain sequence can be a human, or non-human sequence.
- a TCR recognition domain can comprise a MHC polypeptide, a MHC polypeptide sequence, and/or comprise a portion of a MHC sequence.
- the MHC and/or MHC sequence can be autologous, allogeneic, or xenogeneic to a given subject.
- the MHC and/or MHC sequence is a wild-type protein or sequence.
- the MHC and/or MHC sequence is a naturally-occurring variant, e.g., an allele of MHC.
- the MHC and/or MHC sequence is modified relative to a wild-type protein, e.g.
- the MHC and/or MHC sequence is a derivative and/or variant of a wild-type MHC sequence.
- the MHC and/or MHC sequence can be or comprise a human, or non-human sequence.
- the TCR recognition domain comprises a MHC (Major Histocompatibility Complex), a MHC-peptide complex, or a MHC-peptide fusion.
- the TCR recognition domain can comprise a featureless peptide MHC, or a MHC without peptides, or any other molecule that can target or bind specifically to the variable region of the TCR.
- the MHC which is also referred to as the human leukocyte antigen (HLA)
- HLA human leukocyte antigen
- the MHC gene family is divided into three subgroups: MHC class I, MHC class II, and MHC class III.
- Class I MHC molecules have the ⁇ 2 microglobulin subunit which can only be recognised by CD8 co-receptors.
- Class II MHC molecules have ⁇ 1 and ⁇ 2 subunits and can be recognized by CD4 co- receptors.
- MHC molecules chaperone, which type of lymphocytes bind to the given antigen with high affinity, since different lymphocytes express different T-Cell Receptor (TCR) co- receptors.
- TCR T-Cell Receptor
- a complete MHC class I complex comprises one MHC class I heavy chain, one peptide ligand sequence, and a beta 2 microglobulin.
- a TCR recognition domain comprises one MHC class I heavy chain, one peptide ligand sequence, and a beta 2 microglobulin.
- a complete MHC class II complex comprises an MHC class II alpha chain, MHC class II beta chain, and one peptide ligand sequence.
- a TCR recognition domain comprises an MHC class II alpha chain, MHC class II beta chain, and one peptide ligand sequence.
- MHC Major Histocompatibility Complex
- MHC-peptide complex MHC-peptide complex
- MHC-peptide fusion featureless peptide MHC
- MHC-peptide fusion can be selected on the basis of the disease or condition to be treated/prevented.
- Specific MHCs, peptides, and/or antigens that are associated with the diseases described herein are known in the art and an appropriate MHC, peptide, and/or antigen can be selected by one of ordinary skill in the art.
- databases of suitable MHC, peptide, and/or antigen sequences are available on the world wide web at iedb.org; immunespace.org; immgen.org; import/org; peptideatlast.org/repository/; uniprot.org; ncbi.nlm.nih.gov/protein/; immunedata.org/index.php; immuneprofiling.org/hipc/; allergenonline.org/databasebrowe.shtml; and itntrialshare.org.
- TCR recognition domains provides herein are exemplary and non-limiting.
- One of skill in the art can identify relevant auto-antigenic pMHCs, allogeneic peptide MHCs, and autogenic peptide MHCs in addition to those described herein, e.g., from the art and/or from donor cells. Such identification is within the skill of the ordinary practitioner.
- the TCR recognition domain can comprise one or more of SEQ ID NOs: 8-17.
- the TCR recognition domain can comprise SEQ ID NO: 8, 9, and one of 10-17.
- the TCR recognition domain can comprise sequences with at least 80%, at leat 85%, at least 90%, at least 95%, at least 98% or greater sequence identity to SEQ ID NO: 8, 9, and one of 10-17.
- the TCR recognition domain can comprise sequences with at least 95% sequence identity to SEQ ID NO: 8, 9, and one of 10-17, and which retain the wild-type activity of SEQ ID NOs: 8, 9, and one of 10-17.
- SEQ ID NO: 8 HLA-A*0201 (MHC class I heavy chain allele, wildtype, human) MAVMAPRTLVLLLSGALALTQTWAGSHSMRYFFTSVSRPGRGEPRFIAVGYVDDTQFVRFDSDAASQR MEPRAPWIEQEGPEYWDGETRKVKAHSQTHRVDLGTLRGYYNQSEAGSHTVQRMYGCDVGSDWRFLRG YHQYAYDGKDYIALKEDLRSWTAADMAAQTTKHKWEAAHVAEQLRAYLEGTCVEWLRRYLENGKETLQ RTDAPKTHMTHHAVSDHEATLRCWALSFYPAEITLTWQRDGEDQTQDTELVETRPAGDGTFQKWAAVV VPSGQEQRYTCHVQHEGLPKPLTLRWEPSSQPTIPIVGIIAGLVLFGAVITGAVVAAVMWRRKSSDRK GGSYSQAASSDSAQGSDVSLTACKV [0085] SEQ ID NO: 8 H
- the TCR recognition domain can comprise SEQ ID NO: 18, 19, and one of 20-22. In some embodiments, the TCR recognition domain can comprise sequences with at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or greater sequence identity to SEQ ID NO: 18, 19, and one of 20-22. In some embodiments, the TCR recognition domain can comprise sequences with at least 95% sequence identity to SEQ ID NO: 18, 19, and one of 20-22, and which retain the wild-type activity of SEQ ID NOs: 18, 19, and one of 20-22.
- SEQ ID NO: 18 HLA-A*0201 (MHC class I heavy chain allele, wildtype, human, same as previous HLA-A2) MAVMAPRTLVLLLSGALALTQTWAGSHSMRYFFTSVSRPGRGEPRFIAVGYVDDTQFVRFDSDAASQR MEPRAPWIEQEGPEYWDGETRKVKAHSQTHRVDLGTLRGYYNQSEAGSHTVQRMYGCDVGSDWRFLRG YHQYAYDGKDYIALKEDLRSWTAADMAAQTTKHKWEAAHVAEQLRAYLEGTCVEWLRRYLENGKETLQ RTDAPKTHMTHHAVSDHEATLRCWALSFYPAEITLTWQRDGEDQTQDTELVETRPAGDGTFQKWAAVV VPSGQEQRYTCHVQHEGLPKPLTLRWEPSSQPTIPIVGIIAGLVLFGAVITGAVVAAVMWRRKSSDRK GGSYSQAASSDSAQGSDVSLTACKV
- the TCR recognition domain can comprise one or more of the following indicated MHC / peptide pairs, e.g., the TCR recognition domain can comprise one of the indicated MHC alleles and the indicated corresponding peptide. In some embodiments, the TCR recognition domain can comprise sequences with at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or greater sequence identity to one of the following indicated MHC / peptide pairs, e.g., one of the indicated MHC alleles and the indicated corresponding peptide.
- the TCR recognition domain can comprise sequences with at least 95% sequence identity to one of the following indicated MHC / peptide pairs, e.g., one of the indicated MHC alleles and the indicated corresponding peptide, wherein those sequences retain the wild-type activity of the MHC allele and the corresponding peptide .
- Table 5 Each line of the following Table provides a MHC allele (whose sequence is available in publically-accessible databases, e.g., NCBI), and an antigen sequence. The antigen source and/or the relevant disease are also indicated on some lines. In some examples provided in Table 5, the peptides are antigen mimics and their origin is indicated.
- the TCR recognition domain can comprise one or more of the peptides provided in Table 6. In some embodiments, the TCR recognition domain can comprise a sequence with at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or greater sequence identity to one of the peptides of Table 6. In some embodiments, the TCR recognition domain can comprise a sequences with at least 95% sequence identity to one of peptides of Table 6, wherein that sequence retains the wild-type activity of the peptide of Table 6. [00103] Table 6. The sequences are, in the order presented, SEQ ID NOs:1000-2012.
- the peptide is a Minor Histocompatibility Antigen (MiHA).
- MiHAs are known in the art, e.g., in Spierings et al. Tissue Antigens 2014 84:347-360; which is incorporated by reference herein. MiHAs are typically utilized in embodiments relating to transplantation, where it refers to epitopes that are created because of protein sequence variation in polymorphic proteins.
- Residues of the antigenic peptide that engages the MHC binding groove can loosely be divided into two types: anchor residues, which engage with the MHC molecule and confer stability to the MHC-peptide complex (for example, see residues P2, P3, P5, P6, P7, and P9 in Bowness et al. 1999 Expert Reviews in Molecular Medicine 16:1-10; which is incorporated by reference herein in its entirety), and interfacial residues, which are solvent-exposed and can engage with the cognate T-cell receptor (see, e.g., residues P1, P4, and P8 in Bowness).
- anchor residues which engage with the MHC molecule and confer stability to the MHC-peptide complex
- interfacial residues which are solvent-exposed and can engage with the cognate T-cell receptor (see, e.g., residues P1, P4, and P8 in Bowness).
- a featureless peptide is a peptide in which anchor residues are preserved, while interfacial residues of the peptide are mutated to alanine or glycine residues to prevent TCR binding.
- a featureless peptide-MHC is therefore a MHC peptide complex in which the presented peptide is a featureless peptide.
- Featureless peptide MHCs are typically used in embodiments relating to transplantation tolerance. In patients receiving MHC- mismatched solid organ or hematopoietic stem cell transplants, use of a CAL T cell presenting the relevant donor-mismatched featureless peptide-MHC CAL can permit selective depletion of recipient alloreactive T cells targeted towards this mismatched donor HLA allele.
- the MHC can be a monomer, dimer, trimer, tetramer, pentamer, dextramer or other oligomer form.
- the units can be provided in series, e.g., in a chain, or provided arrayed in one or more dimensions around a central point or linker, provided conjugated/bound in any geometry to a scaffold molecule, or any combination of the foregoing.
- TCR recognition domain structures include MHC dimers (Lebowitz et al., 1999 Cellular Immunology 192:175-184), tetramers (Altman et al., 1996 Science 274:94-96), pentamers (proimmune.com/introduction-to-pentamers/), octamers (Guillame et al., 2003 JBC 278:4500-4509), dextramers (Batard et al., 2006 Journal of Immunological Methods 310:136-148), dodecamers (Huang et al., 2016 PNAS 113:E1890-7), lipid vesicles (Mallet-Designe et al., 2003 The Journal of Immunology 170:123-131), and quantum dots (Chattopadhyay et al., 2006 Nature Medicine 12:972- 7).
- the TCR recognition domain can comprise a CD1 domain (e.g., a CD1d domain), e.g., a sequence comprising an extracellular domain of CD1, (e.g., CD1) .
- CD1d domain e.g., a sequence comprising an extracellular domain of CD1, (e.g., CD1) .
- cluster of differentiation 1 family member d” or “CD1d” refers to a cell surface protein that displays lipid antigens to T cells.
- CD1d The sequences of several CD1d isoforms, and the structure of CD1d are known in the art, see, e.g., the 3 isoforms provided in the NCBI database for CD1d (Gene ID 912), and Bagchi et al., 2018 and Oleinika et al., Nature Communcations 20189:684; which are incorporated by reference herein in their entireties.
- isoform 1 of CD1d is SEQ ID NO: 5 (NCBI Ref Seq NP_001757.1)
- isoform 2 of CD1d is SEQ ID NO: 6 (NCBI Ref Seq NP_001306074.1)
- isoform 3 of CD1d is SEQ ID NO: 7 (NCBI Ref Seq NP_001358690.1).
- the CD1d domain comprises a sequence with at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or greater sequence identity to one of SEQ ID NOs: 5-7.
- the CD1d domain comprises a sequence with at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or greater sequence identity to the extracellular domain of one of SEQ ID NOs: 5-7 (e.g., amino acids 20-301 of SEQ ID NO: 5). In some embodiments, the CD1d domain comprises a sequence with at least at least 95% sequence identity to one of SEQ ID NOs: 5-7 and retains the lipid binding activity of the wild-type reference sequence.
- the CD1d domain comprises a sequence with at least 95% sequence identity to the extracellular domain of one of SEQ ID NOs: 5-7 (e.g., amino acids 20-301 of SEQ ID NO: 5) and retains the lipid binding activity of the wild-type reference sequence.
- the CD1 domain further comprises a ligand, e.g., a non-peptide ligand.
- a TCR recognition domain can comprise sequences or molecules in addition to the, e.g., MHC, pMHC, of CD1 domains and sequences.
- it can further comprise other polypeptide and/or non-polypeptide components that enable multimerization.
- Exemplary components that permit multimerization can include biotin (non-polypeptide) and/or streptavidin polypeptide that is used to permit tetramerization.
- protein interaction domains are found on an extracellular portion of the respective polypeptides.
- recognition polypeptide refers to an extracellular polypeptide having a ligand-binding domain.
- the ligand-binding domain can be an antibody reagent.
- the recognition polypeptide can further comprise a protein interaction domain.
- signaling polypeptide refers to a transmembrane polypeptide having an intracellular signaling domain, e.g., a T cell receptor (TCR) signaling domain.
- the signaling polypeptide can further comprise a protein interaction domain.
- the signaling polypeptide can further comprise an extracellular protein interaction domain.
- biomolecular interaction domain refers to a domain that permits specific binding of two separate molecules to each other.
- the molecules can be or can comprise polypeptides. In some embodiments, one or both of the molecules or biomolecular interaction domains can be a non-peptide.
- biomolecular interaction domains When a pair of biomolecular interaction domains is provided herein, they permit two or more molecules to bind specifically, e.g. one of the biomolecular interaction domains can bind specifically to the second biomolecular interaction domain. In some embodiments, specific binding can occur when two separate biomolecular interaction domains, e.g., of a pair, are present. In some embodiments, specific binding can occur when three or more separate biomolecular interaction domains are present. It is noted that protein interaction domains are a type of biomolecular interaction domains and where one is specified herein, the other may always be substituted.
- a molecule when referred to as a protein or polypeptide, it comprises a protein, peptide, or polypeptide sequence, but may comprise additional motifs, modifications, or domains of a non-proteinaceous nature.
- a number of exemplary biomolecular interaction domains, as well as pairs of protein interaction domains are provided elsewhere herein.
- the biomolecular interaction domains comprise, consist, or consist essentially of proteins or polypeptides.
- the biomolecular interaction domains comprise, consist, or consist essentially of non-proteinaceous molecules.
- one of a pair of biomolecular interaction domains can comprise, consist, or consist essentially of proteins or polypeptides and the second of the pair of biomolecular interactions domains can comprise, consist, or consist essentially of a non-proteinaceous molecule (e.g., FITC and anti-FITC).
- FITC and anti-FITC a non-proteinaceous molecule
- Exemplary protein interaction domains are known in the art and can be used in embodiments of the aspects described herein.
- protein interaction domain refers to a domain that permits specific binding of two separate polypeptides to each other. A number of exemplary protein interaction domains, as well as pairs of protein interaction domains are provided elsewhere herein.
- the protein interaction domains of the polypeptides of a multi-component CAL and/or CAR can bind specifically, e.g. one of the protein interaction domains can bind specifically to a second protein interaction domain of the multi-component CAL and/or CAR.
- specific binding can occur when two separate protein interaction domains are present.
- specific binding can occur when three or more separate protein interaction domains are present.
- Exemplary protein interaction domains are known in the art and can be used in embodiments of the aspects described herein. [00117]
- the protein interaction domains can be leucine zipper domains.
- Leucine zipper domains are a type of protein-protein interaction domain commonly found in transcription factors characterized by leucine residues evenly spaced through a ⁇ -helix. Leucine zippers may form heterodimers or homodimers. A number of leucine zipper domains, as well as their ability to bind each other, are known in the art and discussed further, e.g., in Reinke et al. JACS 2010132:6025-31 and Thompson et al. ACS Synth Biol 2012 1:118-129; each of which is incorporated by reference herein in its entirety. In some embodiments, one leucine zipper domain is BZip (RR) and the second leucine zipper domain is AZip (EE).
- RR BZip
- EE AZip
- sequence of a BZip (RR) leucine zipper domain is MDPDLEIRAAFLRQRNTALRTEVAELEQEVQRLENEVSQYETRYGPLGGGK (SEQ ID NO: 3).
- sequence of a AZip (EE) leucine zipper domain is MDPDLEIEAAFLERENTALETRVAELRQRVQRLRNRVSQYRTRYGPLGGGK (SEQ ID NO: 4).
- Further exemplary leucine zipper domains are described in Reinke et al. JACS 2010132:6025-31; which is incorporated by reference herein in its entirety.
- suitable leucine zipper domains can include SYNZIP 1 to SYNZIP 48, and BATF, FOS, ATF4, ATF3, BACH1, JUND, NFE2L3, and HEPTAD. Binding affinities of various combinations of these domains are described, e.g., at Fig.1 of Reinke et al.
- a suitable pair of leucine zipper domains has a dissociation constant (Kd) of 1000 nM or less.
- a suitable pair of leucine zipper domains has a dissociation constant (Kd) of 100 nM or less.
- a suitable pair of leucine zipper domains has a dissociation constant (Kd) of 10 nM or less. In some embodiments, a suitable pair of leucine zipper domains has a dissociation constant (Kd) of 1 nM or less.
- Further exemplary pairs of protein interaction domains can include a) PSD95-Dlg1-zo-1 (PDZ) domains; b) a streptavidin domain and a streptavidin binding protein (SBP) domain; and c) a PYL domain and an ABI domain.
- the protein interaction domains can be chemically-induced protein interaction domains, e.g., domains that will only bind specifically in the presence of a third molecule, e.g., a small molecule or drug.
- Exemplary pairs of chemically-induced protein interaction domains can include: FKBP-binding domain of mTOR (FRB) and FK506 binding protein (FKBP) (binding of which is activated by tacrolimus, everolimus, or a rapalog); cyclophilin-Fas fusion protein (CyP-Fas) and FK506 binding protein (FKBP) (binding of which is activated by FKCsA); calcineurin A (CNA) and FK506 binding protein (FKBP) (binding of which is activated by FK506); gibberellin insensitive (GIA) and gibberellin insensitive dwarf1 (GID1) (binding of which is activated by gibberellin); Snap-tag and Halo tag (binding of which is activated by HaXS); and T14-3-3-cdeltaC and C-Terminal peptides of PMA2 (CT52) (binding of which is activated by fusicoccin).
- the protein interaction domains can comprise at least one nucleotide tag and at least one zinc finger domain.
- Zinc finger domains are characterized by the coordination of a zinc ion in order to stabilize their tertiary structure. The particular folds that appear in zinc fingers can vary.
- a zinc finger domain can be a nucleotide-binding zinc finger domain.
- a zinc finger domain can be a DNA-binding zinc finger domain.
- the protein interaction domain of the recognition polypeptide is a nucleotide tag and the extracellular protein interaction domain of the signaling polypeptide is a zinc finger domain.
- a nucleotide tag can be a DNA tag.
- a nucleotide tag can be a dsDNA tag comprising the entire recognition sequence for the zinc finger domain being used. Exemplary zinc finger domains and their cognate nucleotide tags are described in the art, e.g. Mali et al. Nature Methods 201310:403-406; which is incorporated by reference herein in its entirety.
- a zinc finger domain can be sZF15 as described in Mali et al. Nature Methods 201310:403-406.
- the protein interaction domain(s) is a BZip (RR) and/or a AZip (EE), or any binding pair of protein interaction domains are collectively a BZip (RR) and a AZip (EE).
- the protein interaction domains can comprise a pair of substantially complementary nucleotide tags, e.g., fully complementary or complementary enough to hybridize specifically. The degree of complementarity necessary may vary depending on the total length of the tags and G/C content of the complementary portions.
- a nucleotide tag can be a DNA tag.
- a nucleotide tag can be a DNA tag.
- a nucleotide tag can be a dsDNA tag.
- the protein interaction domain(s) is a gibberellin insensitive (GIA) and/or a gibberellin insensitive dwarf1 (GID1) or any binding pair of protein interaction domains are collectively a gibberellin insensitive (GIA) and a gibberellin insensitive dwarf1 (GID1).
- the protein interaction domain(s) is a Snap-tag and/or a Halo tag, or any binding pair of protein interaction domains are collectively a Snap-tag and a Halo tag.
- the protein interaction domain(s) is a T14-3-3-cdeltaC and/or a C- Terminal peptides of PMA2 (CT52), or any binding pair of protein interaction domains are collectively a T14-3-3-cdeltaC and a C-Terminal peptides of PMA2 (CT52).
- the protein interaction domain(s) is a PYL and/or a ABI, or any binding pair of protein interaction domains are collectively a PYL and a ABI.
- the biomolecular interaction domain(s) is a Fluorescein isothiocyanate (FITC) and/or a FITC binding protein or any binding pair of biomolecular interaction domains are collectively a FITC and a FITC binding protein.
- FITC Fluorescein isothiocyanate
- the biomolecular interaction domain(s) is a (R)-Phycoerythrin (R-PE/ PE) and/or a R-PE/PE binding protein or any binding pair of biomolecular interaction domains are collectively a (R)-Phycoerythrin (R-PE/ PE) and/or a R-PE/PE binding protein.
- the biomolecular domain on the molecule comprising a TCR recognition domain and biomolecular interaction domain i.e. a CAL
- a subject can be treated as described herein without administration of an engineered cell.
- the biomolecular domain on the molecule comprising a TCR recognition domain and biomolecular interaction domain binds specifically to a native cell surface molecule on a NK cell.
- the biomolecular domain on the molecule comprising a TCR recognition domain and biomolecular interaction domain binds specifically to a native cell surface molecule on dendritic cell.
- Exemplary native cell surface molecules for such embodiments include, but are not limited to CD3.
- Other suitable cell surface molecules are the CD cell surface proteins.
- CDs are known in the art and one of ordinary skill can readily select one expressed by the desired cell type(s). For example, see the HCDM database at hcdm.org and the lists available on the world wide web at chemeurope.com/en/encyclopedia/List_of_human_clusters_of_differentiation.html; and docs.abcam.com/pdf/immunology/Guide-to-human-CD-antigens.pdf. Accordingly, exemplary biomolecular interaction domains for such embodiments include but are not limited to an anti-CD3 antibody reagent, or a Fab domain.
- the multiple-component CALs or CARs described herein will activate in the presence of the target ligand, thereby inducing T cell activity in the vicinity of the target ligand.
- multiple-component CALs or CARs capable of logic computation, for example, multiple-component CALs or CARs that serve as AND, OR, or NOT logic gates.
- compositions that comprise components of a multi- component CAL and/or CAR that permits AND gate logic.
- activation of the multi- component CAL and/or CAR happens only in the presence of two target ligands; recognition of a single target ligand is not sufficient for activation.
- Such multi-component CALs or CARs can permit greater specificity and reduce off-target effects. Any single ligand that is a good marker for a target cell or tissue may occur elsewhere in a subject, resulting in off-target effects. However, requiring the recognition of two separate marker ligands reduces the odds of off-target activity.
- a nucleotide tag can be a DNA tag or dsDNA tag.
- a nucleotide tag can be a DNA tag or dsDNA tag.
- the compositions comprise components of a multi-component CAL and/or CAR that are NOT logic gate.
- recognition of a second target ligand by a second recognition polypeptide can prevent interaction (e.g. specific binding) of the signaling polypeptide and first recognition polypeptide.
- Such embodiments can permit suppression of T cell activity in inappropriate and/or off-target tissues.
- the second target ligand can be a marker of a tissue that is particularly sensitive to T cell activity, is a known area of off-target activity, and/or shares markers with the desired target tissue.
- the second target ligand in a NOT gate multi-component CAL and/or CAR, is not a ligand found in the target tissue and/or cells, e.g., in or on a disease T cell.
- the second target ligand of a NOT logic gate multi-component CAL and/or CAR is found on a healthy and/or non-target cell and not on a diseased and/or target cell.
- Various 2- and 3-dimensional configurations of such pairs of nucleotide pairs are known in the art.
- the target ligand recognized by the second recognition polypeptide is found on a healthy and/or non-target cell and not on a diseased and/or target cell.
- the protein interaction domain of the second recognition polypeptide and the protein interaction domain of the first recognition polypeptide have a greater affinity than the protein interaction domain of the signaling polypeptide and the protein interaction domain of the first recognition polypeptide.
- the protein interaction domain of the second recognition polypeptide and the protein interaction domain of the signaling polypeptide have a greater affinity than the protein interaction domain of the signaling polypeptide and the protein interaction domain of the first recognition polypeptide.
- Relative binding affinities can be determined experimentally, e.g., by binding affinity assays known in the art and relative binding affinities are known for a number of combinations of protein interaction domains described herein, see, e.g. Reinke et al. JACS 2010132:6025-31; which is incorporated by reference herein in its entirety.
- the binding affinity of the recognition polypeptide protein interaction domains can be at least 2x greater than the binding affinity of the first recognition polypeptide protein interaction domain and the signaling polypeptide interaction domain.
- the binding affinity of the recognition polypeptide protein interaction domains can be at least 5x greater than the binding affinity of the first recognition polypeptide protein interaction domain and the signaling polypeptide interaction domain.
- the binding affinity of the recognition polypeptide protein interaction domains can be at least 10x greater than the binding affinity of the first recognition polypeptide protein interaction domain and the signaling polypeptide interaction domain.
- target ligand refers to a molecule in or on a cell which can be bound by a ligand-binding domain. Non-limiting examples of such molecules can include polypeptides, lipids, saccharides, and the like. In some embodiments, the target ligand can be an extracellular molecule. In some embodiments, the target ligand can be a cell surface molecule.
- the target ligand (e.g. the first and/or second target ligand) can be a ligand expressed in a target tissue.
- the target ligand can be expressed constitutively in the target tissue and/or cell.
- the target ligand can be expressed exclusively in the target tissue and/or cell.
- the target ligand can be expressed at a higher level in the target tissue and/or cell than in other tissues and/or cells.
- a target ligand in embodiments relating to a multi- component CAL and/or CAR with a single recognition polypeptide or an AND gate multi-component CAL and/or CAR can result in T cell activation (e.g. cell killing activity of the cell comprising the target ligand), the target ligand can be selected to target T cell activity in a desirable and/or therapeutic way, e.g., by targeting a disease cell.
- a target ligand is a ligand found in/on a diseased and/or target cell.
- the target ligand specifically bound by a recognition polypeptide that can specifically bind with a signaling polypeptide or is a portion of an AND gate multi-component CAL and/or CAR is a ligand found in/on a diseased and/or target cell.
- a target ligand specifically bound by a recognition polypeptide that can specifically bind with a signaling polypeptide or is a portion of an AND gate multi-component CAL and/or CAR is a ligand found on a diseased and/or target cell and not on a healthy and/or non-target cell.
- the diseased cell is an autoreactive or alloreactive T cell.
- the target ligand specifically bound by a recognition polypeptide that can specifically bind with a signaling polypeptide or is a portion of an AND gate multi-component CAL and/or CAR is found on the surface of a disease cell.
- a recognition polypeptide that can specifically bind with a signaling polypeptide or is a portion of an AND gate multi-component CAL and/or CAR specifically binds to a target ligand on the surface of a disease cell, e.g. as compared to binding to normal cells.
- a composition and/or cell described herein can further comprise a second multi-component CAL and/or CAR according to any of the aspects and embodiment described herein for the first multi-component CAL and/or CAR.
- a second CAL and/or CAR can be designed to bind specifically to (and, e.g., be activated by or inhibited by) different target ligands than those to which the first multi-component CAL and/or CAR specifically binds (and, e.g. is activated by or inhibited by). This can provide increased specificity, reduced off- target effects, and/or reduced effective dosages for the methods described herein.
- the recognition domain of second multi-component CAL and/or CAR bind specifically to different target ligands than those bound by the recognition domain of the first multi-component CAL and/or CAR.
- the antibody reagents of second multi-component CAL and/or CAR bind specifically to different target ligands than those bound by the antibody reagents of the first multi-component CAL and/or CAR.
- the second multi-component CAL and/or CAR can comprise an inhibitory intracellular signaling domain, e.g., T cell receptor (TCR) signaling domain, e.g., one that inhibits engineered cell, e.g., T cell, activity.
- TCR T cell receptor
- the second multi-component can therefore be designed to operate in opposition to the first multi-component CAL and/or CAR, e.g. permitting inhibition of T cell activation while the first multi-component CAL and/or CAR permits activation of T cell activity.
- Inhibitory intracellular TCR signaling domains are known in the art and can include, by way of non-limiting example, PD1; CTLA4; BTLA; KIR; LAG-3; TIM-3; A2aR; LAIR-1; and TGIT.
- non-active mimetics of an activating TCR can be used.
- the TCR recognition domain comprises a MHC allogeneic to the cell. In one embodiment, the TCR recognition domain comprises a peptide allogeneic to the cell. In one embodiment, the TCR recognition domain comprises a MHC allogeneic to the target cell. In one embodiment, the TCR recognition domain comprises a peptide allogeneic to the target cell. In one embodiment, the TCR recognition domain comprises a non-self peptide relative to the target cell.
- the target ligand specifically bound by a recognition polypeptide that can specifically bind with the signaling polypeptide of the second multi-component CAL and/or CAR comprising an inhibitory intracellular signaling domain is a ligand found on a healthy and/or non-target cell.
- the target ligand specifically bound by a recognition polypeptide that can specifically bind with the signaling polypeptide of the second multi-component CAL and/or CAR comprising an inhibitory intracellular signaling domain is a ligand found on a healthy and/or non-target cell and not on a diseased and/or target cell.
- the second multi-component CAL and/or CAR comprising an inhibitory intracellular signaling domain can be an OR logic gate according to any of the embodiments described herein and the second target ligand can be a ligand found in/on, or specific to, diseased cells.
- a ligand-binding domain can comprise or consist essentially of an antibody reagent.
- the antibody reagent can be an immunoglobulin molecule, a monoclonal antibody, a chimeric antibody, a CDR-grafted antibody, a human antibody, a humanized antibody, a Fab, a Fab', a F(ab')2, a Fv, a disulfide linked Fv, a scFv, a single domain antibody, a diabody, a multispecific antibody, a dual specific antibody, an anti- idiotypic antibody, and/or a bispecific antibody.
- the intracellular signaling domain can be a T-cell activation domain.
- the intracellular signaling domain is a signaling domain from a protein selected from the group consisting of: TCR ⁇ , FcR ⁇ , FcR ⁇ , CD3 ⁇ , CD3 ⁇ , CD3 ⁇ , CD22, CD79a, CD79b, CD66d, CARD11, CD2, CD7, CD27, CD28, CD30, CD40, CD54 (ICAM), CD83, CD134 (OX40), CD137 (4-1BB), CD150 (SLAMF1), CD152 (CTLA4), CD223 (LAG3), CD270 (HVEM), CD273 (PD-L2), CD274 (PD-L1), CD278 (ICOS), DAP10, LAT, NKD2C SLP76, TRIM, and ZAP70.
- a protein selected from the group consisting of: TCR ⁇ , FcR ⁇ , FcR ⁇ , CD3 ⁇ , CD3 ⁇ , CD3 ⁇ , CD22, CD79a, CD79b, CD66d, CARD11, CD2, CD7, CD27,
- the signaling domain can be a paralog or ortholog of any of the foregoing.
- Multi-Component CAR/CAL Cells [00146] Presented herein are cells that express the compositions or multi-component CARs and/or CALs presented herein.
- a cell can be any cell, for example, any mammalian cell, e.g., a human cell.
- the cell is a dendritic cell, regulatory T cell, or effector T cell.
- the cell is a dendritic cell (CAL DC), a T cell (e.g., effector, regulatory, etc.) (CAL-T); regulatory T cell, effector T cell, natural killer cell (CAL NK), or any other myeloid cell.
- an engineered cell expressing and/or comprising one or more multi-component CARs/CALs, or a composition comprising the same as described herein, e.g., at least one signaling polypeptide and at least one recognition polypeptide.
- the cell is a natural killer (NK) cell, dendritic cell, regulatory T cell, effector T cell.
- NK natural killer
- Such cells expressing and/or comprising both a signaling polypeptide and at least one recognition polypeptide of a multi- component CAR/CAL are referred to herein as “complete multi-component CAR/CAL” cells.
- a complete multi-component CAR/CAL cell expresses both a signaling polypeptide (e.g., a CAR) and at least one recognition polypeptide (e.g. a CAL or adaptor as described elsewhere herein) of a multi-component CAL and/or CAR.
- a complete multi- component CAL and/or CAR cell comprises nucleic acid sequences encoding both a signaling polypeptide and at least one recognition polypeptide of a multi-component CAL and/or CAR.
- a signaling polypeptide is present on the membrane of a cell.
- the one or more recognition polypeptides are present in the extracellular space, e.g., the recognition polypeptide(s) can be expressed and secreted by the cell or the cell can be contacted by recognition polypeptides provided from another source (e.g. produced synthetically or by another cell and optionally, purified or processed before the contacting step).
- the recognition and/or signaling polypeptide can be under the control of an inducible and/or repressible promoter. Such promoters allow the expression of the polypeptide to be increased or decreased as desired and are in contrast to constitutive promoters.
- constitutive active promoter refers to a promoter of a gene which is expressed at all times within a given cell.
- exemplary promoters for use in mammalian cells include cytomegalovirus (CMV), Elongation Factor 1a (EF1a), and the like.
- inducible promoter refers to a promoter of a gene which can be expressed in response to a given signal, for example addition or reduction of an agent.
- Non-limiting examples of an inducible promoter are promoters that are regulated in a specific tissue type, a promoter regulated by a steroid hormone, by a polypeptide hormone (e.g., by means of a signal transduction pathway), or by a heterologous polypeptide (e.g., the tetracycline- inducible systems, "Tet-On” and "Tet-Off”; see, e.g., Clontech Inc., CA, Gossen and Bujard, Proc. Natl. Acad. Sci. USA 89:5547, 1992, and Paillard, Human Gene Therapy 9:983, 1989; each of which are incorporated by reference herein in its entirety).
- a heterologous polypeptide e.g., the tetracycline- inducible systems, "Tet-On” and "Tet-Off”; see, e.g., Clontech Inc., CA, Gossen and Bujard, Proc. Natl. Ac
- expression of the polypeptide can be precisely regulated, for example, by using an inducible regulatory sequence that is sensitive to certain physiological regulators, e.g., circulating glucose levels, or hormones (Docherty et al., 1994, FASEB J.8:20-24).
- inducible expression systems suitable for the control of expression in cells or in mammals include, for example, regulation by ecdysone, by estrogen, progesterone, tetracycline, chemical inducers of dimerization, and isopropyl-beta-D1 - thiogalactopyranoside (IPTG).
- IPTG isopropyl-beta-D1 - thiogalactopyranoside
- the expression of one or more of the recognition or signaling polypeptides can be constitutive. In some embodiments, the expression of one or more of the recognition or signaling polypeptides can be transient. Transient expression can be achieved by, e.g., use of transient and/or inducible expression promoters or by use of transient vectors, e.g. those that do not incorporate into the genome and/or persist in the target cell.
- transient vectors e.g. those that do not incorporate into the genome and/or persist in the target cell.
- derivatives of viruses such as the bovine papillomavirus (BPV-1), or Epstein-Barr virus (pHEBo, pREP-derived and p205) can be used for transient expression of nucleic acids in eukaryotic cells.
- the signaling polypeptide of a multi-component CAL and/or CAR can be constitutively expressed and the recognition polypeptide can be transiently expressed.
- the recognition polypeptide of a multi-component CAL and/or CAR can be constitutively expressed and the signaling polypeptide can be transiently expressed.
- the recognition polypeptide of a multi- component CAL can be constitutively expressed and the signaling polypeptide can be provided exogenously. In some embodiments, the recognition polypeptide of a multi- component CAL can be transiently expressed and the signaling polypeptide can be provided exogenously. In some embodiments, the signaling polypeptide of a multi-component CAL and/or CAR can be constitutively expressed and the recognition polypeptide can be provided exogenously. In some embodiments, the signaling polypeptide of a multi-component CAL and/or CAR can be transiently expressed and the recognition polypeptide can be provided exogenously.
- the CARs and CALs described herein can be produced according to any method known in the art, e.g., recombinant expression or peptide synthesis.
- Exemplary methods can include, the NIH Tetramer Core Facility’s MHC expression protocols (available on the world wide web at tetramer.yerkes.emory.edu/support/protocols#1); ProImmune’s pentamer protocols (available on the world wide web at proimmune.com/protocols-2/); Immudex’s dextramer protocols (available on the word wide web at immudex.com/resources/protocols/) and the CAR T cell production protocols provided in the “Primary Human T cells Isolation and Culture” and “Lentiviral Transduction of Human T cells” sub-sections in “Method Details” section of Cho et al., 2018 Cell 173:1426-1438; each of which is incorporated by reference herein in its entirety.
- described herein is a method of killing a target cell, the method comprising contacting the cell with a complete multi-component CAR cell, CAR, and/or CAL according to any of the embodiments described herein.
- the target cell can be a diseased cell, e.g., an autoreactive or alloreactive T cell.
- described herein is a method of treating or preventing a disease, e.g., an autoimmune diseases or conditions; T cell mediated inflammation or immune response; transplant rejection, or GvHD, comprising administering a complete multi- component CAR cell, CAR, and/or CAL according to any of the embodiments described herein.
- described herein is a method of treating or preventing autoimmune diseases or conditions; T cell mediated inflammation or immune response; transplant rejection; or GvHD, comprising administering a complete multi-component CAR cell, CAR, and/or CAL according to any of the embodiments described herein.
- Another aspect provided herein is a method of preventing and/or treating a malignant T cell condition in a subject in need thereof, the method comprising administering to the subject any of the compositions and/or cells described herein.
- described herein is a method of treating or preventing a malignant T cell condition in a subject, comprising administering a complete multi- component CAR/CAL, CAR and/or CAL as according to any of the embodiments described herein.
- described herein is a method of treating or preventing a malignant T cell condition in a subject, comprising administering a complete multi-component CAR cell, CAR, and/or CAL according to any of the embodiments described herein [00153]
- the complete multi-component CAL and/or CAR cell can be autologous or allogeneic to the subject.
- the complete multi-component CAL and/or CAR cell can be derived and/or descended from a cell obtained from the subject or a third party and has been modified ex vivo to comprise the at least one multi-component CAL and/or CAR, e.g., genetically engineered to comprise nucleic acid sequences encoding both a signaling polypeptide and at least one recognition polypeptide of a multi-component CAL and/or CAR.
- the method can further comprise the steps of obtaining a cell from a subject (e.g.
- the engineered cell is further modified to lack or have reduced expression of the native MHCI/II, e.g., as measured on the cell surface.
- Methods for engineering a cell to reduce or eliminate the native MHCI/II include, e.g., expression of RNA interference (such as short hairpin RNA, small interfering RNA, double stranded RNA, etc.), expression of an inhibitory oligonucleotide, nuclease-based inhibition (such as CRISPR, TALEN, Meganuclease, etc.), and expression of a KDEL-motif (SEQ ID NO: 2749) containing binding protein capable of restricting MHCI/II to the endoplasmic reticulum.
- RNA interference such as short hairpin RNA, small interfering RNA, double stranded RNA, etc.
- an inhibitory oligonucleotide such as CRISPR, TALEN, Meganuclease, etc.
- KDEL-motif SEQ ID NO: 2749
- the cell is further engineered to knockout the native MHCI/II.
- Methods for engineering a cell to knockout the native MHCI/II include, e.g., expression of RNA interference (such as short hairpin RNA, small interfering RNA, double stranded RNA, etc.), expression of an inhibitory oligonucleotide, nuclease-based inhibition (such as CRISPR, TALEN, Meganuclease, etc.).
- RNA interference such as short hairpin RNA, small interfering RNA, double stranded RNA, etc.
- an inhibitory oligonucleotide such as CRISPR, TALEN, Meganuclease, etc.
- nuclease-based inhibition such as CRISPR, TALEN, Meganuclease, etc.
- described herein is an engineered cell expressing and/or comprising one or more of the compositions according to any of the embodiments described herein. In one aspect, described herein is an engineered cell expressing and/or comprising one or more multi-component CAL and/or CAR signaling polypeptides according to any of the embodiments described herein. In some embodiments, the cell is a dendritic cell, regulatory T cell, or effector T cell. In some embodiments, the cell is a T cell. Such cells expressing and/or comprising a multi-component CAL and/or CAR signaling polypeptide are referred to herein as “partial multi-component CAL” cells or “partial multi-component CAR” cells.
- the partial multi-component CAL and/or CAR cell does not express, e.g., does not comprise a nucleic acid sequence encoding, a multi- component CAL and/or CAR recognition polypeptide.
- a partial multi- component CAL and/or CAR cell comprises a nucleic acid sequence encoding at least one multi- component CAL and/or CAR signaling polypeptide.
- the multi-component CAL and/or CAR signaling polypeptide is present on the membrane of the cell, e.g., is expressed as a transmembrane protein at detectable levels.
- the signaling polypeptide further comprises a secondary protein interaction domain that specifically binds with the protein interaction domain of the second recognition polypeptide, e.g., the signaling polypeptide is part of an AND gate multi-component CAL and/or CAR as described elsewhere herein.
- the cell can further comprise a second multi-component CAL and/or CAR signaling polypeptide, e.g., a signaling polypeptide that is part of a second multi-component CAL and/or CAR according to any of the embodiments described herein.
- the target cell can be a diseased cell, e.g., an autoreactive or alloreactive T cell.
- the target cell can be a diseased cell, e.g., a cancer cell (e.g., a T cell- or T cell precursor cell- derived neoplasm, hereafter referred to as “T cell neoplasms”.
- the method can further comprise the steps of obtaining a cell from a subject (e.g. a NK cell, a dendritic cell, regulatory T cell, or effector T cell), altering the cell to comprise a nucleic acid sequence encoding a signaling polypeptide of a multi-component CAL and/or CAR, and then administering the cell to the subject.
- a pair of protein interaction domains of a multi-component CAL and/or CAR can comprise chemically induced binding domains and the method can further comprise administering a compound that induces binding of the domains.
- the method when one protein interaction domain is FKBP-binding domain of mTOR (FRB) and a second protein interaction domain is FK506 binding protein (FKBP), the method further comprises administering tacrolimus, a rapalog, or everolimus. In some embodiments, when one protein interaction domain is cyclophilin-Fas fusion protein (CyP-Fas) and a second protein interaction domain is FK506 binding protein (FKBP), the method further comprises administering FKCsA. In some embodiments, when one protein interaction domain is calcineurin A (CNA) and a second protein interaction domain is FK506 binding protein (FKBP), the method further comprises administering FK506.
- FKBP cyclophilin-Fas fusion protein
- FKBP FK506 binding protein
- the method when one protein interaction domain is gibberellin insensitive (GIA) and a second protein interaction domain is gibberellin insensitive dwarf1 (GID1), the method further comprises administering gibberellin. In some embodiments, when one protein interaction domain is Snap-tag and a second protein interaction domain is Halo tag, the method further comprises administering HaXS. In some embodiments, when one protein interaction domain is T14-3-3-cdeltaC and a second protein interaction domain is C-Terminal peptides of PMA2 (CT52), the method further comprises administering fusicoccin. [00159] In some embodiments of any of the aspects described herein, a recognition and/or signaling polypeptide of a multi-component CAL and/or CAR can be engineered.
- a recognition and/or signaling polypeptide of a multi-component CAL and/or CAR can be transgenic. In some embodiments of any of the aspects described herein, a recognition and/or signaling polypeptide of a multi-component CAL and/or CAR can be recombinant. In some embodiments of any of the aspects described herein, a recognition and/or signaling polypeptide of a multi-component CAL and/or CAR can be heterologous to a cell. In some embodiments of any of the aspects described herein, a recognition and/or signaling polypeptide of a multi-component CAL and/or CAR can be heterologous to a T cell.
- a recognition and/or signaling polypeptide of a multi-component CAL and/or CAR can be heterologous to a human T cell. In some embodiments of any of the aspects described herein, a recognition and/or signaling polypeptide of a multi-component CAL and/or CAR can be exogenous to a cell. In some embodiments of any of the aspects described herein, a recognition and/or signaling polypeptide of a multi-component CAL and/or CAR can be exogenous to a T cell.
- a recognition and/or signaling polypeptide of a multi-component CAL and/or CAR can be exogenous to a human T cell.
- each of the individual embodiments described herein can be combined, e.g., in a single cell.
- a single cell could comprise a first complete multi-component CAL and/or CAR and a second partial multi-component CAL and/or CAR, wherein each multi-component CAL and/or CAR can be according to any of the embodiments described herein.
- the methods described herein relate to CAL-immune cell therapies.
- the methods described herein relate to CAR-immune cell therapies such as CAR-T therapy.
- Standard CAR-T and related therapies relate to adoptive cell transfer of immune cells (e.g. T cells) expressing a CAR that binds specifically to a targeted cell type (e.g. disease cells, e.g., autoreactive or alloreactive T cells) to treat a subject, e.g., for an autoimmune diseases or conditions; T cell mediated inflammation or immune response; transplant rejection; or GvHD.
- a targeted cell type e.g. disease cells, e.g., autoreactive or alloreactive T cells
- the cells administered as part of the therapy can be autologous to the subject. In some embodiments, the cells administered as part of the therapy are not autologous to the subject.
- the cells are engineered and/or genetically modified to express a multi-component CAL and/or CAR, or portion thereof as described herein.
- CAR-T therapies can be found, e.g., in Maus et al. Blood 2014123:2624-35; Reardon et al. Neuro- Oncology 2014 16:1441-1458; Hoyos et al. Haematologica 2012 97:1622; Byrd et al. J Clin Oncol 2014 32:3039-47; Maher et al. Cancer Res 2009 69:4559-4562; and Tamada et al. Clin Cancer Res 201218:6436-6445; each of which is incorporated by reference herein in its entirety.
- the technology described herein relates to a syringe or catheter, including an organ-specific catheter (e.g., renal catheter, biliary catheter, cardiac catheter, etc.), comprising a therapeutically effective amount of a composition described herein.
- organ-specific catheter e.g., renal catheter, biliary catheter, cardiac catheter, etc.
- the methods described herein relate to the treatment or prevention of transplant rejection in a subject having a cell, tissue, or organ transplant with one or more compositions, CALs, CARs, or cells as described herein.
- the methods described herein relate to the treatment or prevention of GvHD in a subject having a cell, tissue, or organ transplant with one or more compositions, CALs, CARs, or cells as described herein.
- GvHD refers to a disease characterized by the active process of donor cells attacking the recipient’s own cells. GvHD can develop soon after a transplant, e.g., within weeks or months (acute GvHD), or can occur much later after the transplant, e.g., at least 3-6 months later (chronic GvHD). Symptoms of acute GvHD include, but are not limited to, skin rash or blisters, abdominal pain or discomfort, diarrhea, jaundice, and edema.
- Symptoms of chronic GvHD include, but are not limited to, changes to skin or nail texture, hair loss or thinning, muscle pain or weakness, blurred vision, mouth sores, shortness of breath, persistent cough, abdominal pain or discomfort, and diarrhea.
- a subject can be identified as having or be at risk of having GvHD by a skilled clinician. Diagnostic tests useful in identifying a subject having GvHD are known in the art and will vary based on the type of transplant the subject has received.
- the diagnosis of GvHD is made by, for example, physical examination for the signs and symptoms for GvHD known in the art, serologic testing for dysfunction of the liver, gall bladder, kidney, and hematopoietic cells, histologic analysis of biopsies obtained from affected organs, and radiologic imaging of affected organs.
- the method further comprises administering at least a second therapeutic.
- the composition, CARs, CALs, or cells described herein are administered in combination with Abatacept (Orencia®) or Belatacept (Nulojix®).
- Abatacept and Belatacept are fusion proteins composed of the Fc region of the immunoglobulin IgG1 fused to the extracellular domain of CTLA-4.
- Abatacept is currently approved by the FDA for treatment of rheumatoid arthritis.
- Belatacept which only differs from Abatacept by two amino acids, is an immunosuppressant intended to prevent rejection following a kidney transplant.
- the transplant is vascularized composite allograft (VCA).
- the transplant is any human or non-human cell, tissue, or organ.
- the transplant is any type of transplants procedures, e.g., any heart transplant, any lung transplant, any liver transplant, any pancreas transplant, any cornea transplant, any trachea transplant, any kidney transplant, any skin transplant, any pancreatic islet cell transplant, any allograft (e.g., a transplantation of allogeneic tissue), any xenograft (e.g., a transplantation of xenogeneic tissue), or any autograft (e.g., a transplantation of tissue).
- a skilled practitioner will be able to perform a transplant or identify a subject having had a transplant using standard procedural protocols.
- the methods described herein relate to the treatment or prevention of an autoimmune diseases or conditions, or hypersensitivity reaction I-IV, or immune reaction against foreign therapeutic proteins/molecules, or T cell mediated inflammation or immune response; with compositions, CARs, CALs, or cells as described herein.
- Subjects having an autoimmune disease can be identified by a physician using current methods of diagnosing an autoimmune disease. Symptoms and/or complications of an autoimmune disease which characterize these conditions and aid in diagnosis are well known in the art and include but are not limited to, fatigue, achy muscles, swelling and redness, low-grade fever, numbness or tingling of the hands or feet, hair loss, and/or skin rash.
- autoimmune disease examples include, but are not limited to, blood counts, and an antinuclear antibody test (ANA).
- ANA antinuclear antibody test
- a family history of autoimmune disease, or having risk factors for autoimmune disease e.g. gender, age, ethnicity, and exposure to environmental agents, such as procainamide, hydralazine, mercury, gold, or silver
- risk factors for autoimmune disease e.g. gender, age, ethnicity, and exposure to environmental agents, such as procainamide, hydralazine, mercury, gold, or silver
- autoimmune disease e.g. gender, age, ethnicity, and exposure to environmental agents, such as procainamide, hydralazine, mercury, gold, or silver
- the term "autoimmune disease”, “autoimmune condition”, or “autoimmune disease or disorder” herein is a disease or disorder arising from and directed against an individual's own tissues or a co-segregate or manifestation thereof or resulting condition therefrom.
- Auto-immune related diseases and disorders arise from an overactive and/or abnormal immune response of the body against substances (autoantigens) and tissues normally present in the body, otherwise known as self or autologous substance. This dysregulated inflammatory reaction causes an exaggerated response by macrophages, granulocytes, lymphocytes, and/or T-lymphocytes leading to abnormal tissue damage and cell death. Subsequent loss of function is associated with inflammatory tissue damage.
- Autoantigens as used herein, are endogenous proteins or fragments thereof that are involved in or elicit this pathogenic immune response.
- Autoantigen can be any substance, or a portion thereof normally found within a mammal that, in an autoimmune disease, becomes the primary (or a primary, or secondary) target of attack by the immune system.
- the term also includes antigenic substances that induce conditions having the characteristics of an autoimmune disease when administered to mammals.
- the term includes peptidic subclasses consisting essentially of immunodominant epitopes or immunodominant epitope regions of autoantigens. Immunodominant epitopes or regions in induced autoimmune conditions are fragments of an autoantigen that can be used instead of the entire autoantigen to induce the disease.
- immunodominant epitopes or regions are fragments of antigens specific to the tissue or organ under autoimmune attack and recognized by a substantial percentage (e.g. a majority though not necessarily an absolute majority) of autoimmune attack T-cells.
- Autoantigens that are known to be associated with autoimmune disease include myelin proteins with demyelinating diseases, e.g. multiple sclerosis and experimental autoimmune myelitis; collagens and rheumatoid arthritis; insulin, proinsulin, glutamic acid decarboxylase 65 (GAD65); and islet cell antigen (ICA512; ICA12) for insulin dependent diabetes.
- Th1 type cytokines include interleukin 2 (IL-2), ⁇ -interferon, TNF ⁇ and IL-12.
- cytokines characterized as Th1 type include interleukin 2 (IL-2), interferon ⁇ , and TNF ⁇ .
- IL-2 interleukin 2
- Th1 type include interleukin 2 (IL-2), interferon ⁇ , and TNF ⁇ .
- IL-2 interleukin 2
- Th2 type include IL-10, IL-4 and TGF- ⁇ .
- Th1 and Th2 type T cells may use the identical antigen receptor in response to an immunogen; in the former producing a stimulatory response and, in the latter, a suppressive response.
- T cell mediated inflammation, or a T cell mediated immune response is inflammation and/or an immune response in which T cells and/or T cell activity contributes to or originates the inflammation/immune response.
- inflammation refers to the complex biological response to harmful stimuli, such as pathogens, damaged cells, or irritants. Inflammation is a protective attempt by the organism to remove the injurious stimuli as well as initiate the healing process for the tissue.
- the term “inflammation” includes any cellular process that leads to the production of pro-inflammatory cytokines, inflammation mediators and/or the related downstream cellular events resulting from the actions of the cytokines thus produced, for example, fever, fluid accumulation, swelling, abscess formation, and cell death.
- Inflammation can include both acute responses (i.e., responses in which the inflammatory processes are active) and chronic responses (i.e., responses marked by slow progression and formation of new connective tissue).
- Acute and chronic inflammation may be distinguished by the cell types involved. Acute inflammation often involves polymorphonuclear neutrophils; whereas chronic inflammation is normally characterized by a lymphohistiocytic and/or granulomatous response.
- An inflammatory condition is any disease state characterized by inflammatory tissues (for example, infiltrates of leukocytes such as lymphocytes, neutrophils, macrophages, eosinophils, mast cells, basophils and dendritic cells) or inflammatory processes which provoke or contribute to the abnormal clinical and histological characteristics of the disease state.
- inflammatory tissues for example, infiltrates of leukocytes such as lymphocytes, neutrophils, macrophages, eosinophils, mast cells, basophils and dendritic cells
- an “immune response” refers to a response by a cell of the immune system, such as a B cell, T cell (CD4 or CD8), regulatory T cell, antigen-presenting cell, dendritic cell, monocyte, macrophage, NKT cell, NK cell, basophil, eosinophil, or neutrophil, to a stimulus (e.g., to an a disease, an antigen, or healthy cells, e.g., in the case of autoimmunity).
- a T cell response such as a CD4+ response or a CD8+ response.
- Such responses by these cells can include, for example, cytotoxicity, proliferation, cytokine or chemokine production, trafficking, or phagocytosis, and can be dependent on the nature of the immune cell undergoing the response.
- Stimulation of an immune response refers to an induction or increase of the immune response.
- Suppression of an immune response refers to an elimination or decrease of the immune response.
- a "cell-mediated immune response” is one mediated by T-cells and/or other white blood cells.
- a “cell-mediated immune response” is elicited by the presentation of antigenic epitopes in association with Class I or Class II molecules of the major histocompatibility complex (MHC), CD1 or other non-classical MHC-like molecules.
- MHC major histocompatibility complex
- CTLs have specificity for peptide antigens that are presented in association with proteins encoded by classical or non-classical MHCs and expressed on the surfaces of cells. CTLs help induce and promote the intracellular destruction of intracellular microbes, or the lysis of cells infected with such microbes.
- Another aspect of cellular immunity involves an antigen-specific response by helper T-cells. Helper T-cells act to help stimulate the function, and focus the activity of, nonspecific effector cells against cells displaying peptide or other antigens in association with classical or non-classical MHC molecules on their surface.
- a “cell- mediated immune response” also refers to the production of cytokines, chemokines and other such molecules produced by activated T-cells and/or other white blood cells, including those derived from CD4+ and CD8+ T-cells.
- the stimulation of a cell-mediated immunological response may be determined by a number of assays, such as by lymphoproliferation (lymphocyte activation) assays, CTL cytotoxic cell assays, by assaying for T-lymphocytes specific for the antigen in a sensitized subject, or by measurement of cytokine production by T cells in response to re-stimulation with antigen.
- assays are well known in the art. See, e.g., Erickson et al.
- the T cell mediated immune response is a response to a drug administered to the subject. It is contemplated herein that the present technology can be utilized for depletion of anti-drug specific T cells to prevent immune responses against administered biologics, cell therapies, and/or gene therapies.
- the methods and compositions described herein can be used to prevent or treat anti-AAV and anti-transgene immune responses for administered adeno- associated virus (AAV) gene therapies, preventing or treating immune to responses to genome editing agents such as CRISPR/Cas9, Transcription activator-like effector nucleases (TALENs), or Zinc Finger Nucleases (ZFNs), and preventing or treating immune responses to enzyme replacement therapies such as recombinant human acid ⁇ -glucosidase (Pompe disease), ⁇ -L-iduronidase (Mucopolysaccharidosis I), and ⁇ -galactosidase (Fabry disease).
- AAV adeno- associated virus
- the autoimmune disorder is selected from the group consisting of thyroiditis, type 1 diabetes mellitus, Hashimoto's thyroiditis, Graves' disease, celiac disease, multiple sclerosis, Guillain-Barre syndrome, Addison's disease, and Raynaud's phenomenon, Goodpasture's disease, arthritis (rheumatoid arthritis such as acute arthritis, chronic rheumatoid arthritis, gout or gouty arthritis, acute gouty arthritis, acute immunological arthritis, chronic inflammatory arthritis, degenerative arthritis, type II collagen-induced arthritis, infectious arthritis, Lyme arthritis (e.g., post treatment Lyme disease syndrome), proliferative arthritis, psoriatic arthritis, Still's disease, vertebral arthritis, and juvenile-onset rheumatoid arthritis, arthritis chronica progrediente, arthritis deformans, polyarthritis chronica primaria, reactive arthritis, and ankylosing spondylitis),
- arthritis rheumatoid arthritis such as
- the autoimmune disease or condition or T cell mediated inflammation can be neurodegeneration, e.g, Alzheimer’s or Parksinson disease.
- the autoimmune disease or condition, or T cell mediated inflammation can be type 1 diabetes, rheumatoid arthritis, multiple sclerosis, pemphigus, alopecia, lupus, vitiligo, or chronic fatigue syndrome.
- malignant T cell condition refers to a condition in which T cells display one or more of uncontrolled growth (i.e., division beyond normal limits), invasion (i.e., intrusion on and destruction of adjacent tissues), and metastasis (i.e., spread to other locations in the body via lymph or blood).
- Non-limiting examples of malignant T cell conditions include T cell cancers, lymphoma, leukemia, T cell acute lymphoblastic leukemia, and T cell lymphoblastic lymphoma.
- the compositions and methods described herein can be administered to a subject to treat or prevent an autoimmune diseases or conditions; T cell mediated inflammation or immune response; or transplant rejection.
- the methods described herein comprise administering an effective amount of compositions, CALs, CARs, or cells described herein to a subject in order to alleviate a symptom of an autoimmune diseases or conditions; T cell mediated inflammation or immune response; or transplant rejection.
- "alleviating a symptom” is ameliorating any condition or symptom associated with the disease or condition. As compared with an equivalent untreated control, such reduction is by at least 5%, 10%, 20%, 40%, 50%, 60%, 80%, 90%, 95%, 99% or more as measured by any standard technique.
- a variety of means for administering the compositions described herein to subjects are known to those of skill in the art.
- compositions contemplated herein may be carried out in any convenient manner, including by aerosol inhalation, injection, ingestion, transfusion, implantation or transplantation. In a preferred embodiment, compositions are administered parenterally.
- parenteral administration and “administered parenterally” as used herein refers to modes of administration other than enteral and topical administration, usually by injection, and includes, without limitation, intravascular, intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intratumoral, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcuticular, intraarticular, subcapsular, subarachnoid, intraspinal and intrasternal injection and infusion.
- the compositions contemplated herein are administered to a subject by direct injection into a tumor, lymph node, or site of infection.
- a pharmaceutical composition comprising the cells, e.g., T cells or CAL cells or CAR cells, described herein may be administered at a dosage of 10 2 to 10 10 cells/kg body weight, preferably 10 5 to 10 6 cells/kg body weight, including all integer values within those ranges.
- the number of cells will depend upon the ultimate use for which the composition is intended as will the type of cells included therein.
- the cells are generally in a volume of a liter or less, can be 500 mLs or less, even 250 mLs or 100 mLs or less.
- the density of the desired cells is typically greater than 10 6 cells/ml and generally is greater than 10 7 cells/ml, generally 10 8 cells/ml or greater.
- the clinically relevant number of immune cells can be apportioned into multiple infusions that cumulatively equal or exceed 10 5 , 10 6 , 10 7 , 10 8 , 10 9 , 10 10 , 10 11 , or 10 12 cells.
- lower numbers of cells in the range of 10 6 /kilogram (10 6 -10 11 per patient) may be administered.
- CAL and/or CAR expressing cell compositions may be administered multiple times at dosages within these ranges.
- the cells may be allogeneic, syngeneic, xenogeneic, or autologous to the patient undergoing therapy.
- the treatment may also include administration of mitogens (e.g., PHA) or lymphokines, cytokines, and/or chemokines (e.g., IFN- ⁇ , IL-2, IL-12, TNF-alpha, IL-18, and TNF-beta, GM-CSF, IL-4, IL-13, Flt3- L, RANTES, MIP1 ⁇ , etc.) as described herein to enhance induction of the immune response.
- mitogens e.g., PHA
- lymphokines e.g., lymphokines, cytokines, and/or chemokines (e.g., IFN- ⁇ , IL-2, IL-12, TNF-alpha, IL-18, and TNF-beta, GM-CSF, IL-4, IL-13, Flt3- L, RANTES, M
- the dosage can be from about 1x10 6 cells to about 1x10 7 cells per kg of body weight. In some embodiments, the dosage can be about 1x10 6 cells per kg of body weight. In some embodiments, one dose of cells can be administered. In some embodiments, the dose of cells can be repeated, e.g., once, twice, or more. In some embodiments, the dose of cells can be administered on, e.g., a daily, weekly, or monthly basis. [00187]
- the dosage ranges for the agent, e.g., a CAL, CAR, cell, or composition described herein depend upon the potency, and encompass amounts large enough to produce the desired effect e.g., prevention of transplant rejection, reduction in inflammation, etc.
- the dosage should not be so large as to cause unacceptable adverse side effects.
- the dosage will vary with the age, condition, and sex of the patient and can be determined by one of skill in the art.
- the dosage can also be adjusted by the individual physician in the event of any complication.
- the dosage ranges from 0.001 mg/kg body weight to 0.5 mg/kg body weight.
- the dose range is from 5 ⁇ g/kg body weight to 100 ⁇ g/kg body weight.
- the dose range can be titrated to maintain serum levels between 1 ⁇ g/mL and 1000 ⁇ g/mL.
- subjects can be administered a therapeutic amount, such as, e.g., 0.1 mg/kg, 0.5 mg/kg, 1.0 mg/kg, 2.0 mg/kg, 2.5 mg/kg, 5 mg/kg, 10 mg/kg, 15 mg/kg, 20 mg/kg, 25 mg/kg, 30 mg/kg, 40 mg/kg, 50 mg/kg, or more.
- Administration of the doses recited above can be repeated.
- the doses are given once a day, or multiple times a day, for example but not limited to three times a day.
- the doses recited above are administered daily for several weeks or months. The duration of treatment depends upon the subject’s clinical progress and responsiveness to therapy.
- the dose can be from about 2 mg/kg to about 15 mg/kg. In some embodiments, the dose can be about 2 mg/kg. In some embodiments, the dose can be about 4 mg/kg. In some embodiments, the dose can be about 5 mg/kg. In some embodiments, the dose can be about 6 mg/kg. In some embodiments, the dose can be about 8 mg/kg. In some embodiments, the dose can be about 10 mg/kg. In some embodiments, the dose can be about 15 mg/kg. In some embodiments, the dose can be from about 100 mg/m 2 to about 700 mg/m 2 . In some embodiments, the dose can be about 250 mg/m 2 . In some embodiments, the dose can be about 375 mg/m 2 .
- the dose can be about 400 mg/m 2 . In some embodiments, the dose can be about 500 mg/m 2 .
- the dose can be administered intravenously. In some embodiments, the intravenous administration can be an infusion occurring over a period of from about 10 minutes to about 3 hours. In some embodiments, the intravenous administration can be an infusion occurring over a period of from about 30 minutes to about 90 minutes. [00191] In some embodiments the dose can be administered about weekly. In some embodiments, the dose can be administered weekly. In some embodiments, the dose can be administered weekly for from about 12 weeks to about 18 weeks. In some embodiments the dose can be administered about every 2 weeks. In some embodiments the dose can be administered about every 3 weeks.
- the dose can be from about 2 mg/kg to about 15 mg/kg administered about every 2 weeks. In some embodiments, the dose can be from about 2 mg/kg to about 15 mg/kg administered about every 3 weeks. In some embodiments, the dose can be from about 2 mg/kg to about 15 mg/kg administered intravenously about every 2 weeks. In some embodiments, the dose can be from about 2 mg/kg to about 15 mg/kg administered intravenously about every 3 weeks. In some embodiments, the dose can be from about 200 mg/m2 to about 400 mg/m2 administered intravenously about every week. In some embodiments, the dose can be from about 200 mg/m2 to about 400 mg/m2 administered intravenously about every 2 weeks.
- the dose can be from about 200 mg/m2 to about 400 mg/m2 administered intravenously about every 3 weeks. In some embodiments, a total of from about 2 to about 10 doses are administered. In some embodiments, a total of 4 doses are administered. In some embodiments, a total of 5 doses are administered. In some embodiments, a total of 6 doses are administered. In some embodiments, a total of 7 doses are administered. In some embodiments, a total of 8 doses are administered. In some embodiments, the administration occurs for a total of from about 4 weeks to about 12 weeks. In some embodiments, the administration occurs for a total of about 6 weeks. In some embodiments, the administration occurs for a total of about 8 weeks.
- the administration occurs for a total of about 12 weeks.
- the initial dose can be from about 1.5 to about 2.5 fold greater than subsequent doses.
- the dose can be from about 1 mg to about 2000 mg.
- the dose can be about 3 mg.
- the dose can be about 10 mg.
- the dose can be about 30 mg.
- the dose can be about 1000 mg.
- the dose can be about 2000 mg.
- the dose can be about 3 mg given by intravenous infusion daily.
- the dose can be about 10 mg given by intravenous infusion daily.
- the dose can be about 30 mg given by intravenous infusion three times per week.
- a therapeutically effective amount is an amount of an agent that is sufficient to produce a statistically significant, measurable change in, or prevent the occurrence of an autoimmune disease or condition; T cell mediated inflammation or immune response; transplant rejection; or GvHD. Such effective amounts can be gauged in clinical trials as well as animal studies.
- An agent can be administered intravenously by injection or by gradual infusion over time.
- agents useful in the methods and compositions described herein can be administered intravenously, intranasally, by inhalation, intraperitoneally, intramuscularly, subcutaneously, intracavity, and can be delivered by peristaltic means, if desired, or by other means known by those skilled in the art. It is preferred that the compounds used herein are administered orally, intravenously or intramuscularly. Local administration, e.g., directly to the site of an organ or tissue transplant is also specifically contemplated. [00195] Therapeutic compositions containing at least one agent can be conventionally administered in a unit dose, for example.
- unit dose when used in reference to a therapeutic composition refers to physically discrete units suitable as unitary dosage for the subject, each unit containing a predetermined quantity of active material calculated to produce the desired therapeutic effect in association with the required physiologically acceptable diluent, i.e., carrier, or vehicle.
- the compositions are administered in a manner compatible with the dosage formulation, and in a therapeutically effective amount.
- the quantity to be administered and timing depends on the subject to be treated, capacity of the subject’s system to utilize the active ingredient, and degree of therapeutic effect desired.
- the partial multi-component CAL and/or CAR cell and a recognition polypeptide can be administered together or separately.
- each of the compositions can be administered, separately, according to any of the dosages and administration routes/routines described herein.
- Precise amounts of active ingredient required to be administered depend on the judgment of the practitioner and are particular to each individual. However, suitable dosage ranges for systemic application are disclosed herein and depend on the route of administration.
- Suitable regimes for administration are also variable but are typified by an initial administration followed by repeated doses at one or more hour intervals by a subsequent injection or other administration.
- continuous intravenous infusion sufficient to maintain concentrations in the blood in the ranges specified for in vivo therapies are contemplated.
- the methods further comprise administering a composition, CAL, or CAR, or cell described herein along with one or more additional autoimmune, GvHD, or transplant rejection agents, biologics, drugs, or treatments as part of a combinatorial therapy.
- Exemplary treatments for transplant rejection or GvHD include but are not limited to, Immunosuppressive drugs, e.g., Cyclosporine (Neoral, Sandimmune, Gengraf, and Restasis), Tacrolimus (Prograf, Protopic, Astagraf XL, and Envarsus XR), Methotrexate (Trexall, Rasuvo, Rheumatrex, and Otrexup (PF)), Sirolimus (Rapamune), Mycophenolic acid (Myfortic and CellCept), Rituximab (Rituxan), etanercept (Enbrel), pentostatin (Nipent), ruxolitinib (Jakafi); Chemotherapies, e.g., Methotrexate (Trexall, Rasuvo, Rheumatrex, and Otrexup (PF)), antithymocyte globulin (Atgam, Thymoglobulin); Steroids, e.g,.,
- Prednisone (Deltasone, Rayos, and Prednisone Intensol), Methylprednisolone (Medrol, Solu- Medrol, and Depo-Medrol), budesonide (Entocort EC, Uceris); Antifungal, e.g., Posaconazole (Noxafil); Antiviral drugs, e.g., Acyclovir (Zovirax and Sitavig), Valacyclovir (Valtrex); and Antibiotics, e.g., Sulfamethoxazole / Trimethoprim (Bactrim, Sulfatrim, and Bactrim DS); Protease inhibitors, e.g.
- alpha1-proteinase inhibitor Zemaira
- extracorporeal photopheresis monoclonal antibodies
- diaclizumab Zainbryta
- basiliximab Simulect
- Brentuximab vedotin Adcetris
- Alemtuzumab Campath, Lemtrada
- Tocilizumab Actemra
- infusion of mesenchymal stromal cells infusion of mesenchymal stromal cells.
- Exemplary treatments for autoimmune disease include but are not limited to, Insulin, e.g., Insulin glulisine (Apidra and Apidra SoloStar), Insulin detemir (Levemir and Levemir FlexTouch), Insulin aspart (NovoLog, Novolog Flexpen, and Novolog PenFill), Insulin lispro (Humalog and Humalog KwikPen), Insulin, Insulin glargine (Lantus, Lantus Solostar, and Toujeo SoloStar); Dietary supplement, e.g., glucose tablets; and Hormones, e.g., Glucagon (GlucaGen and Glucagon Emergency Kit (human)), antidiabetic agents (Metformin (D-Care DM2, Fortamet, Glucophage, Glucophage XR, Glumetza, Riomet), glucagon-like peptide-1 (GLP-1) receptor agonist (liraglutide (Saxenda) Insulin
- a treatment is considered “effective treatment,” as the term is used herein, if any one or all of the signs or symptoms of are altered in a beneficial manner or other clinically accepted symptoms are improved, or even ameliorated, e.g., by at least 10% following treatment with an agent as described herein. Efficacy can also be measured by a failure of an individual to worsen as assessed by hospitalization or need for medical interventions (i.e., progression of the disease is halted). Methods of measuring these indicators are known to those of skill in the art and/or described herein. [00202] An effective amount for the treatment of a disease means that amount which, when administered to a mammal in need thereof, is sufficient to result in effective treatment as that term is defined herein, for that disease.
- Efficacy of an agent can be determined by assessing physical indicators of, for example autoimmune disease (e.g., result of an ANA), T cell mediated inflammation or immune response, malignant T cell condition, transplant rejection (e.g., high fever, tenderness at transplant site, etc.), or GvHD (e.g., redness, pain, or other symptoms at transplant site).
- Effective amounts, toxicity, and therapeutic efficacy can be determined by standard pharmaceutical procedures in cell cultures or experimental animals, e.g., for determining the LD50 (the dose lethal to 50% of the population) and the ED50 (the dose therapeutically effective in 50% of the population).
- the dosage can vary depending upon the dosage form employed and the route of administration utilized.
- the dose ratio between toxic and therapeutic effects is the therapeutic index and can be expressed as the ratio LD50/ED50.
- Compositions and methods that exhibit large therapeutic indices are preferred.
- a therapeutically effective dose can be estimated initially from cell culture assays.
- a dose can be formulated in animal models to achieve a circulating plasma concentration range that includes the IC50 (i.e., the concentration of the active ingredient(s), which achieves a half-maximal inhibition of symptoms) as determined in cell culture, or in an appropriate animal model.
- Levels in plasma can be measured, for example, by immunoassay, various DNA detection technologies, or high performance liquid chromatography.
- any particular dosage can be monitored by a suitable bioassay, e.g., assay to assess reaction following transplant, level of inflammation, ANA measurement, among others.
- the dosage can be determined by a physician and adjusted, as necessary, to suit observed effects of the treatment.
- Efficacy can also be measured by a failure of an individual to worsen as assessed by hospitalization, or need for medical interventions (i.e., progression of the disease is halted). Methods of measuring these indicators are known to those of skill in the art and/or are described herein.
- Treatment includes any treatment of a disease in an individual or an animal (some non-limiting examples include a human or an animal) and includes: (1) inhibiting the disease, e.g., preventing a worsening of symptoms (e.g. pain or inflammation); or (2) relieving the severity of the disease, e.g., causing regression of symptoms.
- An effective amount for the treatment of a disease means that amount which, when administered to a subject in need thereof, is sufficient to result in effective treatment as that term is defined herein, for that disease.
- Efficacy of an agent can be determined by assessing physical indicators of a condition or desired response, (e.g. a reduction of inflammation, etc.).
- Efficacy can be assessed in animal models of a condition described herein, for example treatment of autoimmune disease, transplant rejection or GVHD.
- efficacy of treatment is evidenced when a statistically significant change in a marker is observed, e.g. inflammation.
- the technology described herein relates to a pharmaceutical composition
- a pharmaceutical composition comprising a CAL and/or CAR, or a multi-component CAL and/or CAR (or portion thereof, or cell comprising a CAL and/or CAR, or a multi-component CAL and/or CAR) as described herein, and optionally a pharmaceutically acceptable carrier.
- the active ingredients of the pharmaceutical composition comprise a CAL and/or CAR or a multi-component CAL and/or CAR (or portion thereof, or cell comprising a CAL and/or CAR or a multi-component CAL and/or CAR) as described herein.
- the active ingredients of the pharmaceutical composition consist essentially of a CAL and/or CAR or a multi-component CAL and/or CAR (or portion thereof, or cell comprising a CAL and/or CAR or a multi-component CAL and/or CAR) as described herein. In some embodiments, the active ingredients of the pharmaceutical composition consist of a CAL and/or CAR or a multi-component CAL and/or CAR (or portion thereof, or cell comprising a multi-component CAL and/or CAR) as described herein. [00205] Pharmaceutically acceptable carriers and diluents include saline, aqueous buffer solutions, solvents and/or dispersion media.
- Some non-limiting examples of materials which can serve as pharmaceutically-acceptable carriers include: (1) sugars, such as lactose, glucose and sucrose; (2) starches, such as corn starch and potato starch; (3) cellulose, and its derivatives, such as sodium carboxymethyl cellulose, methylcellulose, ethyl cellulose, microcrystalline cellulose and cellulose acetate; (4) powdered tragacanth; (5) malt; (6) gelatin; (7) lubricating agents, such as magnesium stearate, sodium lauryl sulfate and talc; (8) excipients, such as cocoa butter and suppository waxes; (9) oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; (10) glycols, such as propylene glycol; (11) polyols, such as glycerin, sorbitol, mannito
- the pharmaceutical composition comprising a multi-component CAL and/or CAR (or portion thereof, or cell comprising a multi-component CAL and/or CAR) as described herein can be a parenteral dose form.
- parenteral dosage forms are preferably sterile or capable of being sterilized prior to administration to a patient.
- parenteral dosage forms include, but are not limited to, solutions ready for injection, dry products ready to be dissolved or suspended in a pharmaceutically acceptable vehicle for injection, suspensions ready for injection, and emulsions.
- controlled-release parenteral dosage forms can be prepared for administration of a patient, including, but not limited to, DUROS®-type dosage forms and dose-dumping.
- Suitable vehicles that can be used to provide parenteral dosage forms of a multi- component CAL and/or CAR (or portion thereof, or cell comprising a multi-component CAL and/or CAR) as disclosed within are well known to those skilled in the art.
- Examples include, without limitation: sterile water; water for injection USP; saline solution; glucose solution; aqueous vehicles such as but not limited to, sodium chloride injection, Ringer's injection, dextrose Injection, dextrose and sodium chloride injection, and lactated Ringer's injection; water-miscible vehicles such as, but not limited to, ethyl alcohol, polyethylene glycol, and propylene glycol; and non-aqueous vehicles such as, but not limited to, corn oil, cottonseed oil, peanut oil, sesame oil, ethyl oleate, isopropyl myristate, and benzyl benzoate.
- aqueous vehicles such as but not limited to, sodium chloride injection, Ringer's injection, dextrose Injection, dextrose and sodium chloride injection, and lactated Ringer's injection
- water-miscible vehicles such as, but not limited to, ethyl alcohol, polyethylene glycol, and propylene
- compositions can also be formulated to be suitable for oral administration, for example as discrete dosage forms, such as, but not limited to, tablets (including without limitation scored or coated tablets), pills, caplets, capsules, chewable tablets, powder packets, cachets, troches, wafers, aerosol sprays, or liquids, such as but not limited to, syrups, elixirs, solutions or suspensions in an aqueous liquid, a non-aqueous liquid, an oil-in-water emulsion, or a water-in-oil emulsion.
- compositions contain a predetermined amount of the pharmaceutically acceptable salt of the disclosed compounds, and may be prepared by methods of pharmacy well known to those skilled in the art. See generally, Remington: The Science and Practice of Pharmacy, 21st Ed., Lippincott, Williams, and Wilkins, Philadelphia PA. (2005).
- Conventional dosage forms generally provide rapid or immediate drug release from the formulation. Depending on the pharmacology and pharmacokinetics of the drug, use of conventional dosage forms can lead to wide fluctuations in the concentrations of the drug in a patient's blood and other tissues. These fluctuations can impact a number of parameters, such as dose frequency, onset of action, duration of efficacy, maintenance of therapeutic blood levels, toxicity, side effects, and the like.
- controlled-release formulations can be used to control a drug's onset of action, duration of action, plasma levels within the therapeutic window, and peak blood levels.
- controlled- or extended-release dosage forms or formulations can be used to ensure that the maximum effectiveness of a drug is achieved while minimizing potential adverse effects and safety concerns, which can occur both from under-dosing a drug (i.e., going below the minimum therapeutic levels) as well as exceeding the toxicity level for the drug.
- the composition can be administered in a sustained release formulation.
- Controlled-release pharmaceutical products have a common goal of improving drug therapy over that achieved by their non-controlled release counterparts.
- the use of an optimally designed controlled-release preparation in medical treatment is characterized by a minimum of drug substance being employed to cure or control the condition in a minimum amount of time.
- Advantages of controlled-release formulations include: 1) extended activity of the drug; 2) reduced dosage frequency; 3) increased patient compliance; 4) usage of less total drug; 5) reduction in local or systemic side effects; 6) minimization of drug accumulation; 7) reduction in blood level fluctuations; 8) improvement in efficacy of treatment; 9) reduction of potentiation or loss of drug activity; and 10) improvement in speed of control of diseases or conditions.
- controlled-release formulations are designed to initially release an amount of drug (active ingredient) that promptly produces the desired therapeutic effect, and gradually and continually release other amounts of drug to maintain this level of therapeutic or prophylactic effect over an extended period of time. In order to maintain this constant level of drug in the body, the drug must be released from the dosage form at a rate that will replace the amount of drug being metabolized and excreted from the body.
- Controlled-release of an active ingredient can be stimulated by various conditions including, but not limited to, pH, ionic strength, osmotic pressure, temperature, enzymes, water, and other physiological conditions or compounds.
- a variety of known controlled- or extended-release dosage forms, formulations, and devices can be adapted for use with the salts and compositions of the disclosure.
- dosage forms can be used to provide slow or controlled-release of one or more active ingredients using, for example, hydroxypropylmethyl cellulose, other polymer matrices, gels, permeable membranes, osmotic systems (such as OROS® (Alza Corporation, Mountain View, Calif.
- CAL-cell therapy seeks to help immune killer cells recognize autoreactive and alloreactive T cells. This is accomplished by genetically altering an immune cell so that it expresses a chimeric antigen ligand (CAL).
- CAL is an altered ligand, in which the natural recognition portion is removed and replaced with a synthetic recognition portion, (including all synthetic or natural peptide MHC complexes) that is designed to more effectively recognize the autoreactive and alloreactive T cells by very specifically detecting the presence of a T cell receptor unique to the autoreactive and alloreactive T cells.
- This invention is based, in part, on the finding that an engineered polypeptide presented herein can recognize and bind to the specific T cell receptor on the disease-causing T cells, deleting said T cells, e.g., eliminating or reducing autoimmune diseases or conditions; T cell mediated inflammation or immune response; and cell, tissue, organ transplants and Graft vs. Host Disease (GvHD).
- the engineered polypeptide is composed of a peptide-major histocompatibility complex (pMHC) (e.g. as a monomer, oligomer, or multimer) as the recognition site for the TCR of an allogeneic or an autoreactive T cell.
- pMHC peptide-major histocompatibility complex
- the pMHC is one of the complexes described herein, e.g., see Tables 5 and 6.
- the engineered polypeptide is composed of a pMHC conjugated to a FITC, PE, or other biomolecular interaction domain described herein.
- the engineered polypeptide can also be considered as an adaptor molecule (referred to herein as Chimeric Antigen Ligand (CAL)) composed of a TCR recognition domain (e.g., peptide-HLA monomers, oligomers, or multimers) fused to a biomolecular interaction domain.
- CAL Chimeric Antigen Ligand
- the CAL technology described herein can target T cell clones in an antigen specific manner and is not dependent on any specific CAR construct to exhibit killing effect.
- the engineered polypeptide is a CAR composed of a peptide-HLA (e.g., as a monomer, oligomer, or multimer) as the recognition site fused to signaling domains from T cell receptors.
- a split version of this CAR system in which the CAR is composed of two pieces.
- the first piece is a universal CAR (also referred to herein as a Uni CAL) with T cell signaling domains as the intracellular portion and a biomolecular interaction domain as the extracellular domain, e.g., that is specific to each disease state.
- the second piece is an adaptor molecule (referred to herein as Chimeric Antigen Ligand (CAL)) composed of a TCR recognition domain (e.g., peptide-HLA monomers, oligomers, or multimers) fused to the cognate biomolecular interaction domain.
- CAL Chimeric Antigen Ligand
- the engineered polypeptide is a CAR composed of a peptide-HLA (e.g., as a monomer, oligomer, or multimer) as the recognition site fused to signaling domains from T cell receptors. Further provided herein is a split version of this CAR system, in which the CAR is composed of two pieces.
- the first piece is a universal CAR (also referred to herein as a Uni CAL) with T cell signaling domains as the intracellular portion and a biomolecular interaction domain as the extracellular domain, e.g., that is specific to each disease state.
- the second piece is an adaptor molecule (referred to herein as Chimeric Antigen Ligand (CAL)) composed of a TCR recognition domain (e.g., peptide-HLA monomers, oligomers, or multimers) fused to the cognate biomolecular interaction domain.
- CAL Chimeric Antigen Ligand
- protein interaction domains are a type of biomolecular interaction domains and where one is specified herein, the other may always be substituted.
- one aspect presented herein provides a composition comprising (a) chimeric antigen ligand or a TCR recognition domain; and one or both of (a) an intracellular signaling domain; and (b) a first-type protein interaction domain.
- compositions comprising (a) a first polypeptide comprising a chimeric antigen ligand or TCR recognition domain and a first-type protein interaction domain; and (b) a signaling polypeptide comprising a second-type protein interaction domain and an intracellular signaling domain; wherein the first-type and second-type protein interaction domains bind specifically to each other.
- compositions comprising (a) a first polypeptide comprising a TCR recognition domain and a first-type protein interaction domain; and (b) a recognition polypeptide comprising a second recognition domain and a third-type protein interaction domain; wherein the first-type and third-type protein interaction domains bind specifically to each other.
- compositions comprising (a) a first polypeptide comprising a TCR recognition domain and a first-type protein interaction domain; and (b) a signaling polypeptide comprising a second-type protein interaction domain and an intracellular signaling domain; and (c) a recognition polypeptide comprising a second recognition domain and a third-type protein interaction domain; wherein the second-type and third-type protein interaction domains compete for binding to the first-type protein interaction domain.
- the third-type protein interaction domain and first-type protein interaction domain have a higher affinity for each other than the second-type protein interaction domain and first-type protein interaction domain.
- compositions comprising (a) a first polypeptide comprising a TCR recognition domain and a first-type protein interaction domain; (b) a signaling polypeptide comprising a second-type protein interaction domain, a fourth-type protein interaction domain, and an intracellular signaling domain; and (c) a recognition polypeptide comprising a second recognition domain and a fifth-type protein interaction domain; wherein the first-type protein interaction domain and the second-type protein interaction domain bind specifically to each other; and wherein the fourth-type protein interaction domain and the fifth-type protein interaction domain bind specifically to each other.
- the fourth-type protein interaction domain and fifth- type protein interaction domain have a weaker affinity than the second-type protein interaction domain and first-type protein interaction domain.
- the first polypeptide further comprises a sixth-type protein interaction domain and the recognition polypeptide further comprises a seventh-type protein interaction domain which bind specifically to each other.
- the second recognition domain is specific for a target that is not recognized by the TCR recognition domain.
- the second recognition domain is specific for a target that is found on a healthy and/or non-target cell and not on a diseased and/or target cell.
- TCR recognition domains may not comprise a single polypeptide, but rather comprise two or more polypeptides and may additionally even comprise non-polypeptides.
- a single MHC class II tetramer TCR recognition domain can comprise 4 biotin small molecules and 16 polypeptides (4 peptides, 4 MHC class II alpha chains, 4 MHC class II beta chains, and 4 streptavidin proteins.
- Other types of TCR recognition domains may additionally comprise other non-polypeptide molecules (e.g., MHC dextramers contain a polysaccharide backbone to which the MHCs are anchored to).
- a composition described herein can comprise multiple copies or instances of a TCR recognition domain(s), e.g.
- the TCR recognition domain can be a mulitmer, or oligomer.
- a composition described herein can comprise multiple copies or instances of a first polypeptide as described herein.
- the first polypeptide comprises the entire TCR recognition domain.
- the TCR recognition domain comprises at least two separate polypeptide sequences, the first polypeptide comprises at least one of the separate polypeptide sequences of the TCR recognition domain, and the first polypeptide is bound to or complexed with a second or further polypeptide sequences of the TCR recognition domain to form a TCR recognition domain.
- the TCR recognition domain comprises a MHC (Major Histocompatibility Complex); a MHC-peptide complex; or a MHC-peptide fusion.
- the peptide is a human, non-human, or synthetic/engineered peptide.
- the peptides can further comprise non-proteinaceous motifs, modifications, or domains, e.g., they can comprise glycosylation and/or lipids.
- the peptide is a Minor Histocompatibility Antigen (MiHA).
- the MHC is a monomer, dimer, trimer, tetramer, pentamer, dextramer or other oligomer form.
- the protein interaction domains are found on an extracellular portion of the respective polypeptides.
- the protein interaction domain(s) is a leucine zipper, or any binding pair of protein interaction domains are collectively a pair of leucine zippers;
- the protein interaction domain(s) is a BZip (RR) and/or a AZip (EE), or any binding pair of protein interaction domains are collectively a BZip (RR) and a AZip (EE);
- the protein interaction domain(s) is a PSD95-Dlgl-zo-1 (PDZ) domain;
- the protein interaction domain(s) is a streptavidin and/or a streptavidin binding protein (SBP) or any binding pair of protein interaction domains are collectively a streptavidin and a streptavidin binding protein (SBP);
- the protein interaction domain(s) is a FKBP-binding domain of mTOR (FRB) and/or a FK506 binding protein (FKBP) or any binding pair
- the nucleotide tag is a DNA tag or dsDNA tag.
- the intracellular signaling domain is a signaling domain from a protein selected from the group consisting of: TCR ⁇ , FcR ⁇ , FcR ⁇ , CD3 ⁇ ; CD35; CD3 ⁇ ; CD3C; CD22; CD79a; CD79b; CD66d; CARD11; CD2; CD7; CD27; CD28; CD30; CD40; CD54 (ICAM); CD83; CD134 (OX40); CD137 (4-1BB); CD150 (SLAMF1); CD152 (CTLA4); CD223 (LAG3); CD270 (HVEM); CD273 (PD-L2); CD274 (PD- Ll); CD278 (ICOS); DAP10; LAT; KD2C SLP76; TRIM; and ZAP70.
- a cell comprising and/or expressing a comprising a composition comprising a TCR recognition domain and an intracellular signaling domain further comprises a TCR signaling-responsive promoter operatively linked to a payload transgene.
- a TCR signaling-responsive promoter operatively linked to a payload transgene.
- Such embodiments permit transgene payload expression specifially to and/or in the vicinity of a targeted T cell.
- Suitable promoters and transgene are known in the art, e.g., those promoters and transgenes used in “TRUCK CAR” technology.
- An exemplary promoter is a NFAT-sensitive promoter.
- transgene payloads can include checkpoint inhibitors (e.g., CTLA-4, [Ipilimumab, Tremelimumab] or PD-1 [Nivolumab, Pembrolizumab, Pidilizumab]) or proinflammatory cytokines (e.g., IL-2, IL-12, etc).
- checkpoint inhibitors e.g., CTLA-4, [Ipilimumab, Tremelimumab] or PD-1 [Nivolumab, Pembrolizumab, Pidilizumab]
- proinflammatory cytokines e.g., IL-2, IL-12, etc.
- this will be used for in-situ targeting of a patient’s anticancer T-cells to very specifically and locally deliver activating agents like one or more checkpoint inhibitors (CTLA-4, [Ipilimumab, Tremelimumab] or PD-1 [Nivolumab, Pembrolizumab, Pidilizumab]) and/or proinflammatory cytokines, (e.g. IL-2, IL-12, etc.) that can push them to expansion and effector phenotype.
- CTL-4 checkpoint inhibitors
- PD-1 PD-1
- proinflammatory cytokines e.g. IL-2, IL-12, etc.
- the cell can be allogeneic, e.g., and engineered once and given as pulse therapy.
- the cell can be a T cell or any other cell type described herein, e.g., a NK cell.
- exemplary, non-limiting proinflammatory cytokines include IFNs, IFN- ⁇ , TNF ⁇ , TGF- ⁇ , IL-1 ⁇ , IL-6, IL-4, IL-10, IL-13, IL-2, IL-12, IL-15, and IL-27.
- a promoter can be said to drive expression or drive transcription of the nucleic acid sequence that it regulates.
- a composition comprising a TCR recognition domain and an biomolecular interaction domain is a soluble molecule and/or soluble complex.
- Another aspect provided herein is a cell comprising and/or expressing the composition of any of the compositions described herein.
- the TCR recognition domain comprises a MHC allogeneic to the cell comprising and/or expressing the composition.
- the TCR recognition domain comprises a MHC allogeneic to the cell that the TCR originated from.
- the TCR recognition domain comprises a peptide allogeneic to the cell comprising and/or expressing the composition.
- the TCR recognition domain comprises a peptide allogeneic to the cell that the TCR originated from.
- the cell is a dendritic cell (CAL DC), a T cell (e.g., effector, regulatory, etc.,) (CAL-T); regulatory T cell, effector T cell, natural killer cell (CAL NK), or any other myeloid cell.
- the cell is engineered to express the polypeptide(s) of the composition. In one embodiment of any aspect, the cell is engineered to express the signaling polypeptide of the composition. In one embodiment of any aspect, the cell is further engineered to knockout the native MHCI/II. In one embodiment of any aspect, the cell is further engineered to lack cell surface expression of native MHCI/II.
- CAR chimeric antigen receptor
- Another aspect provided herein is a chimeric antigen receptor (CAR) comprising (a) an anti-CD127 and/or anti-CD45RO recognition domain; and (b) an intracellular signaling domain.
- composition comprising a first polypeptide comprising: (a) an anti-CD127 and/or anti-CD45RO recognition domain; (b) a first-type protein interaction domain; and a second polypeptide comprising (a) a second-type protein interaction domain; and (b) an intracellular signaling domain; wherein the first-type protein interaction domain and the second-type protein interaction domain bind specifically to each other.
- composition comprising a first polypeptide comprising (a) an anti-CD127 recognition domain; (b) a first-type protein interaction domain; a second polypeptide comprising (a) an anti-CD45RO recognition domain; (b) a fifth-type protein interaction domain; and a third polypeptide comprising (a) a second-type and a fourth-type protein interaction domain; and (b) an intracellular signaling domain; wherein the first-type protein interaction domain and the second-type protein interaction domain bind specifically to each other; and wherein the fourth-type protein interaction domain and the fifth-type protein interaction domain bind specifically to each other.
- Another aspect provided herein is a cell comprising any of the CARs described herein, or any of the compositions described herein.
- Another aspect provided herein is a method of preventing and/or treating an autoimmune diseases or conditions or T cell mediated inflammation or immune response; or treating or preventing transplant rejection or GvHD in a subject in need thereof, the method comprising administering to the subject any of the compositions and/or cells described herein.
- Another aspect provided herein is a method of preventing and/or treating a malignant T cell condition in a subject in need thereof, the method comprising administering to the subject any of the compositions and/or cells described herein.
- the TCR recognition domain comprises a MHC allogeneic to the subject, a MHC autologous to the transplant cells, a peptide allogeneic to the subject, or a peptide autologous to the transplant cells.
- the transplant is vascularized composite allotransplantation (VCA).
- VCA vascularized composite allotransplantation
- the autoimmune disease is type 1 diabetes, multiple sclerosis, rheumatoid arthritis, or scleroderma.
- One aspect of the embodiments provided herein is a CAR T cell that targets a CD127+/CD45RO+ T cell.
- the CD127+/CD45RO+ T cell is a CD127+/CD45RO+ memory T cell. In one embodiment, the CD127+/CD45RO+ T cell is an alloreactive CD127+/CD45RO+ T cell. [00252] Accordingly, one aspect herein provides a CAR comprising (a) an anti-CD127 and/or anti-CD45RO recognition domain; and (b) intracellular signaling domain. Further provided herein is a composition comprising (a) an anti-CD127 and/or anti-CD45RO recognition domain; and (b) intracellular signaling domain.
- composition comprising a first polypeptide comprising: (a) an anti-CD127 and/or anti-CD45RO recognition domain; (b) a first-type protein interaction domain; and a second polypeptide comprising: (c) a second-type protein interaction domain; and (d) an intracellular signaling domain; wherein the first-type protein interaction domain and the second- type protein interaction domain bind specifically to each other.
- composition comprising a first polypeptide comprising: (a) an anti-CD127 recognition domain; (b) a first-type protein interaction domain; a second polypeptide comprising: (c) an anti-CD45RO recognition domain; (d) a fifth-type protein interaction domain; and a third polypeptide comprising: (e) a second-type and a fourth-type protein interaction domain; and (f) an intracellular signaling domain; wherein the first-type protein interaction domain and the second-type protein interaction domain bind specifically to each other; and wherein the fourth-type protein interaction domain and the fifth-type protein interaction domain bind specifically to each other.
- the anti-CD127 recognition domain recognizes and binds to the sequence of CD127 on a CD127+ cell, e.g., a CD127+/CD45RO+ T cell.
- CD127 also referred to as Interleukin 7 receptor a (IL7Ra), ILRA, IL7RA, CDW127, or IL-7R-alpha
- IL7Ra Interleukin 7 receptor a
- ILRA Interleukin 7 receptor a
- CDW127 CDW127
- IL-7R-alpha is a cell surface receptor that has been shown to have a role in V(D)J recombination during lymphocyte development. Defects in CD127 have been associated with severe combined immunodeficiency (SCID).
- SCID severe combined immunodeficiency
- CD127 refers to all naturally occurring variants or isoforms of CD127.
- the CD127 polypeptide sequence is presented in SEQ ID NO: 1.
- the CD127 polypeptide can be an ortholog, variant, and/or allele of SEQ ID NO: 1.
- CD45RO also referred to as PTPRC, LCA, LY5, B220, CD45, L-CA, T200, CD45R, and GP180, is to a cell surface signaling molecule that been shown to be an essential regulator of T- and B-cell antigen receptor signaling. Sequences for CD45RO are known for a number of species, e.g., human CD45RO (NCBI Gene ID: 5788), mRNA (NCBI Ref Seq: NM_001267798.2), and polypeptide (NCBI Ref Seq: NP_001254727.1). CD45RO refers to all naturally occurring variants or isoforms of CD45RO.
- the CD45RO polypeptide sequence is presented in SEQ ID NO: 2.
- the CD45RO polypeptide can be an ortholog, variant, and/or allele of SEQ ID NO: 2.
- “reduce,” “reduction” or “decrease” or “inhibit” typically means a decrease by at least 10% as compared to a reference level (e.g. the absence of a given treatment) and can include, for example, a decrease by at least about 10%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about 99% , or more.
- “reduction” or “inhibition” does not encompass a complete inhibition or reduction as compared to a reference level. “Complete inhibition” is a 100% inhibition as compared to a reference level. A decrease can be preferably down to a level accepted as within the range of normal for an individual without a given disorder. [00260] The terms “increased”, “increase”, “enhance”, or “activate” are all used herein to mean an increase by a statically significant amount.
- the terms “increased”, “increase”, “enhance”, or “activate” can mean an increase of at least 10% as compared to a reference level, for example an increase of at least about 20%, or at least about 30%, or at least about 40%, or at least about 50%, or at least about 60%, or at least about 70%, or at least about 80%, or at least about 90% or up to and including a 100% increase or any increase between 10-100% as compared to a reference level, or at least about a 2-fold, or at least about a 3-fold, or at least about a 4-fold, or at least about a 5-fold or at least about a 10-fold increase, or any increase between 2-fold and 10-fold or greater as compared to a reference level.
- a “increase” is a statistically significant increase in such level.
- a "subject” means a human or animal. Usually the animal is a vertebrate such as a swine, primate, rodent, domestic animal or game animal. Primates include chimpanzees, cynomolgus monkeys, spider monkeys, and macaques, e.g., Rhesus. Rodents include mice, rats, woodchucks, ferrets, rabbits and hamsters.
- domestic and game animals include cows, horses, pigs, deer, bison, buffalo, feline species, e.g., domestic cat, canine species, e.g., dog, fox, wolf, avian species, e.g., chicken, emu, ostrich, and fish, e.g., trout, catfish and salmon.
- the subject is a mammal, e.g., a primate, e.g., a human.
- the terms, “individual,” “patient” and “subject” are used interchangeably herein. [00262]
- the subject is a mammal.
- the mammal can be a human, non-human swine, primate, mouse, rat, dog, cat, horse, or cow, but is not limited to these examples. Mammals other than humans can be advantageously used as subjects that represent animal models of autoimmune diseases or conditions; T cell mediated inflammation or immune response; malignant T cell condition; transplant rejection; or GvHD.
- a subject can be male or female.
- a subject can be one who has been previously diagnosed with or identified as suffering from or having a condition in need of treatment (e.g.
- an autoimmune disease or condition T cell mediated inflammation or immune response; malignant T cell condition; transplant rejection; or GvHD
- T cell mediated inflammation or immune response T cell mediated inflammation or immune response
- malignant T cell condition transplant rejection
- GvHD GvHD
- a subject can also be one who has not been previously diagnosed as having an autoimmune disease or condition; T cell mediated inflammation or immune response; malignant T cell condition; transplant rejection; or GvHD or one or more complications related to an autoimmune disease or condition; T cell mediated inflammation or immune response; malignant T cell condition; transplant rejection; or GvHD.
- a subject can be one who exhibits one or more risk factors for an autoimmune disease or condition; T cell mediated inflammation or immune response; malignant T cell condition; transplant rejection; or GvHD or one or more complications related to an autoimmune disease or condition; T cell mediated inflammation or immune response; malignant T cell condition; transplant rejection; or GvHD or a subject who does not exhibit risk factors.
- a “subject in need” of treatment for a particular condition can be a subject having that condition, diagnosed as having that condition, or at risk of developing that condition, e.g., an autoimmune disease or condition; T cell mediated inflammation or immune response; malignant T cell condition; transplant rejection; or GvHD.
- a nucleic acid encoding a CAL, CAR, a multi-component CAL and/or CAR or portion thereof as described herein is comprised by a vector.
- a nucleic acid sequence encoding a multi-component CAL and/or CAR, or portion thereof as described herein, or any module thereof is operably linked to a vector.
- vector refers to a nucleic acid construct designed for delivery to a host cell or for transfer between different host cells.
- a vector can be viral or non-viral.
- vector encompasses any genetic element that is capable of replication when associated with the proper control elements and that can transfer gene sequences to cells.
- a vector can include, but is not limited to, a cloning vector, an expression vector, a plasmid, phage, transposon, cosmid, chromosome, virus, virion, etc.
- expression vector refers to a vector that directs expression of an RNA or polypeptide from sequences linked to transcriptional regulatory sequences on the vector. The sequences expressed will often, but not necessarily, be heterologous to the cell.
- An expression vector may comprise additional elements, for example, the expression vector may have two replication systems, thus allowing it to be maintained in two organisms, for example in human cells for expression and in a prokaryotic host for cloning and amplification.
- expression refers to the cellular processes involved in producing RNA and proteins and as appropriate, secreting proteins, including where applicable, but not limited to, for example, transcription, transcript processing, translation and protein folding, modification and processing.
- “Expression products” include RNA transcribed from a gene, and polypeptides obtained by translation of mRNA transcribed from a gene.
- gene means the nucleic acid sequence that is transcribed (DNA) to RNA in vitro or in vivo when operably linked to appropriate regulatory sequences.
- the gene may or may not include regions preceding and following the coding region, e.g.5′ untranslated (5′UTR) or “leader” sequences and 3′ UTR or “trailer” sequences, as well as intervening sequences (introns) between individual coding segments (exons).
- the term “viral vector” refers to a nucleic acid vector construct that includes at least one element of viral origin and has the capacity to be packaged into a viral vector particle.
- the viral vector can contain the nucleic acid encoding a CAL and/or CAR described herein, e.g., a multi-component CAL and/or CAR, or portion thereof as described herein in place of non- essential viral genes.
- the vector and/or particle may be utilized for the purpose of transferring any nucleic acids into cells either in vitro or in vivo.
- Numerous forms of viral vectors are known in the art.
- recombinant vector is meant a vector that includes a heterologous nucleic acid sequence, or “transgene” that is capable of expression in vivo or in the transduced cells. It should be understood that the vectors described herein can, in some embodiments, be combined with other suitable compositions and therapies.
- the vector is episomal. The use of a suitable episomal vector provides a means of maintaining the nucleotide of interest in the subject in high copy number extra chromosomal DNA thereby eliminating potential effects of chromosomal integration.
- nucleic acid or “nucleic acid sequence” refers to any molecule, preferably a polymeric molecule, incorporating units of ribonucleic acid, deoxyribonucleic acid or an analog thereof.
- the nucleic acid can be either single-stranded or double-stranded.
- a single-stranded nucleic acid can be one nucleic acid strand of a denatured double- stranded DNA. Alternatively, it can be a single-stranded nucleic acid not derived from any double-stranded DNA.
- the nucleic acid can be DNA.
- the nucleic acid can be RNA, e.g., single-stranded or double-stranded RNA.
- Suitable nucleic acid molecules are DNA, including genomic DNA or cDNA.
- Other suitable nucleic acid molecules are RNA, including mRNA.
- protein and “polypeptide” are used interchangeably herein to designate a series of amino acid residues, connected to each other by peptide bonds between the alpha-amino and carboxy groups of adjacent residues.
- protein refers to a polymer of amino acids, including modified amino acids (e.g., phosphorylated, glycated, glycosylated, etc.) and amino acid analogs, regardless of its size or function.
- modified amino acids e.g., phosphorylated, glycated, glycosylated, etc.
- amino acid analogs regardless of its size or function.
- Protein and polypeptide are often used in reference to relatively large polypeptides, whereas the term “peptide” is often used in reference to small polypeptides, but usage of these terms in the art overlaps.
- protein and “polypeptide” are used interchangeably herein when referring to a gene product and fragments thereof.
- exemplary polypeptides or proteins include gene products, naturally occurring proteins, homologs, orthologs, paralogs, fragments and other equivalents, variants, fragments, and analogs of the foregoing.
- an “antibody” refers to IgG, IgM, IgA, IgD or IgE molecules or antigen- specific antibody fragments thereof (including, but not limited to, a Fab, F(ab')2, Fv, disulphide linked Fv, scFv, single domain antibody, closed conformation multispecific antibody, disulphide-linked scfv, diabody), whether derived from any species that naturally produces an antibody, or created by recombinant DNA technology; whether isolated from serum, B-cells, hybridomas, transfectomas, yeast or bacteria.
- an "antigen” is a molecule that is bound by a binding site on an antibody agent.
- antigens are bound by antibody ligands and are capable of raising an antibody response in vivo.
- An antigen can be a polypeptide, protein, nucleic acid or other molecule or portion thereof.
- the term “antigenic determinant” refers to an epitope on the antigen recognized by an antigen-binding molecule, and more particularly, by the antigen-binding site of said molecule.
- antibody reagent refers to a polypeptide that includes at least one immunoglobulin variable domain or immunoglobulin variable domain sequence and which specifically binds a given antigen.
- An antibody reagent can comprise an antibody or a polypeptide comprising an antigen-binding domain of an antibody.
- an antibody reagent can comprise a monoclonal antibody or a polypeptide comprising an antigen-binding domain of a monoclonal antibody.
- an antibody can include a heavy (H) chain variable region (abbreviated herein as VH), and a light (L) chain variable region (abbreviated herein as VL).
- an antibody includes two heavy (H) chain variable regions and two light (L) chain variable regions.
- antibody reagent encompasses antigen-binding fragments of antibodies (e.g., single chain antibodies, Fab and sFab fragments, F(ab')2, Fd fragments, Fv fragments, scFv, and domain antibodies (dAb) fragments (see, e.g. de Wildt et al., Eur J. Immunol.1996; 26(3):629-39; which is incorporated by reference herein in its entirety)) as well as complete antibodies.
- An antibody can have the structural features of IgA, IgG, IgE, IgD, IgM (as well as subtypes and combinations thereof).
- Antibodies can be from any source, including mouse, rabbit, pig, rat, and primate (human and non-human primate) and primatized antibodies. Antibodies also include midibodies, humanized antibodies, chimeric antibodies, and the like. [00274]
- the VH and VL regions can be further subdivided into regions of hypervariability, termed “complementarity determining regions” ("CDR"), interspersed with regions that are more conserved, termed “framework regions” ("FR").
- CDR complementarity determining regions
- FR framework regions
- Each VH and VL is typically composed of three CDRs and four FRs, arranged from amino-terminus to carboxy-terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4.
- FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4 The terms "antigen-binding fragment” or “antigen-binding domain”, which are used interchangeably herein are used to refer to one or more fragments of a full length antibody that retain the ability to specifically bind to a target of interest.
- binding fragments encompassed within the term "antigen-binding fragment" of a full length antibody include (i) a Fab fragment, a monovalent fragment consisting of the VL, VH, CL and CH1 domains; (ii) a F(ab')2 fragment, a bivalent fragment including two Fab fragments linked by a disulfide bridge at the hinge region; (iii) an Fd fragment consisting of the VH and CH1 domains; (iv) an Fv fragment consisting of the VL and VH domains of a single arm of an antibody, (v) a dAb fragment (Ward et al., (1989) Nature 341:544- 546; which is incorporated by reference herein in its entirety), which consists of a VH or VL domain; and (vi) an isolated complementarity determining region (CDR) that retains specific antigen-binding functionality.
- CDR complementarity determining region
- specific binding refers to a chemical interaction between two molecules, compounds, cells and/or particles wherein the first entity binds to the second, target entity with greater specificity and affinity than it binds to a third entity which is a non-target.
- specific binding can refer to an affinity of the first entity for the second target entity which is at least 10 times, at least 50 times, at least 100 times, at least 500 times, at least 1000 times or greater than the affinity for the third nontarget entity.
- a reagent specific for a given target is one that exhibits specific binding for that target under the conditions of the assay being utilized.
- binding described herein can be preferential binding, e.g., binding between two molecules, compounds, cells and/or particles wherein the first entity binds to the second, target entity with at least 2 times greater specificity and affinity than it binds to a third entity which is a non-target.
- a recombinant humanized antibody can be further optimized to decrease potential immunogenicity, while maintaining functional activity, for therapy in humans.
- functional activity means a polypeptide capable of displaying one or more known functional activities associated with a recombinant antibody or antibody reagent thereof as described herein. Such functional activities include, e.g. the ability to bind to a target.
- the terms “treat,” “treatment,” “treating,” or “amelioration” refer to therapeutic treatments, wherein the object is to reverse, alleviate, ameliorate, inhibit, slow down or stop the progression or severity of a condition associated with a disease or disorder, e.g. autoimmune disease or condition; T cell mediated inflammation or immune response; malignant T cell condition; transplant rejection; or GvHD.
- the term “treating” includes reducing or alleviating at least one adverse effect or symptom of a condition, disease or disorder associated with a, e.g. autoimmune disease or condition; T cell mediated inflammation or immune response; malignant T cell condition; transplant rejection; or GvHD. Treatment is generally “effective” if one or more symptoms or clinical markers are reduced.
- treatment is “effective” if the progression of a disease is reduced or halted. That is, “treatment” includes not just the improvement of symptoms or markers, but also a cessation of, or at least slowing of, progress or worsening of symptoms compared to what would be expected in the absence of treatment.
- Beneficial or desired clinical results include, but are not limited to, alleviation of one or more symptom(s), diminishment of extent of disease, stabilized (i.e., not worsening) state of disease, delay or slowing of disease progression, amelioration or palliation of the disease state, remission (whether partial or total), and/or decreased mortality, whether detectable or undetectable.
- treatment also includes providing relief from the symptoms or side-effects of the disease (including palliative treatment).
- pharmaceutical composition refers to the active agent in combination with a pharmaceutically acceptable carrier e.g. a carrier commonly used in the pharmaceutical industry.
- pharmaceutically acceptable is employed herein to refer to those compounds, materials, compositions, and/or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit/risk ratio.
- administering refers to the placement of an agent, e.g., a CAL, CAR, composition, or cell as disclosed herein into a subject by a method or route which results in at least partial delivery of the agent at a desired site.
- Pharmaceutical compositions comprising the compounds disclosed herein can be administered by any appropriate route which results in an effective treatment in the subject.
- Immune checkpoint inhibitors inhibit one or more immune checkpoint proteins. The immune system has multiple inhibitory pathways that are critical for maintaining self-tolerance and modulating immune responses.
- immune checkpoint protein refers to a protein which, when active, exhibits an inhibitory effect on immune activity, e.g., T cell activity.
- Exemplary immune checkpoint proteins can include PD-1 (e.g., NCBI Gene ID: 5133); PD-L1 (e.g., NCBI Gene ID: 29126); PD-L2 (e.g., NCBI Gene ID: 80380); TIM-3 (e.g., NCBI Gene ID: 84868); CTLA4 (e.g., NCBI Gene ID: 1493); TIGIT (e.g., NCBI Gene ID: 201633); KIR (e.g., NCBI Gene ID: 3811); LAG3 (e.g., NCBI Gene ID: 3902); DD1- ⁇ (e.g., NCBI Gene ID: 64115); A2AR (e.g., NCBI Gene ID: 135); B7-H3 (e.g., NCBI Gene ID: 80381); B7-H4 (e.g., NCBI Gene ID: 79679); BTLA (e.g., NCBI Gene ID: 151888); IDO (e.g., NCBI Gene ID
- B7 family ligands include, but are not limited to, B7- 1, B7-2, B7-DC, B7-H1, B7-H2, B7-H3, B7-H4, B7-H5, B7-H6 and B7-H7.
- Non-limiting examples of immune checkpoint inhibitors can include:MGA271 (B7-H3: MacroGenics); ipilimumab (CTLA-4; Bristol Meyers Squibb); pembrolizumab (PD-1; Merck); nivolumab (PD-1; Bristol Meyers Squibb) ; atezolizumab (PD-L1; Genentech); galiximab (B7.1; Biogen); IMP321 (LAG3: Immuntep); BMS-986016 (LAG3; Bristol Meyers Squibb); SMB-663513 (CD137; Bristol-Meyers Squibb); PF- 05082566 (CD137; Pfizer); IPH2101 (KIR; Innate Pharma); KW-0761 (CCR4; Kyowa Kirin); CDX- 1127 (CD27; CellDex); MEDI-6769 (Ox40;
- compositions, methods, and respective component(s) thereof are essential to the method or composition, yet open to the inclusion of unspecified elements, whether essential or not.
- consisting of refers to compositions, methods, and respective components thereof as described herein, which are exclusive of any element not recited in that description of the embodiment.
- consisting essentially of refers to those elements required for a given embodiment. The term permits the presence of elements that do not materially affect the basic and novel or functional characteristic(s) of that embodiment.
- a composition comprising: a. a TCR recognition domain; and one or both of: b. an intracellular signaling domain; and c. a first-type protein interaction domain.
- a composition comprising: a. a first polypeptide comprising a TCR recognition domain and a first-type protein interaction domain; and b.
- a composition comprising: a. a first polypeptide comprising a TCR recognition domain and a first-type protein interaction domain; and b. a recognition polypeptide comprising a second recognition domain and a third-type protein interaction domain; wherein the first-type and third-type protein interaction domains bind specifically to each other.
- a composition comprising: a. a first polypeptide comprising a TCR recognition domain and a first-type protein interaction domain; and b.
- a composition comprising: a. a first polypeptide comprising a TCR recognition domain and a first-type protein interaction domain; b.
- a signaling polypeptide comprising a second-type protein interaction domain, a fourth-type protein interaction domain, and an intracellular signaling domain; and c. a recognition polypeptide comprising a second recognition domain and a fifth-type protein interaction domain; wherein the first-type protein interaction domain and the second-type protein interaction domain bind specifically to each other; and wherein the fourth-type protein interaction domain and the fifth-type protein interaction domain bind specifically to each other. 7. The composition of paragraph 6, wherein the fourth-type protein interaction domain and fifth- type protein interaction domain have a weaker affinity than the second-type protein interaction domain and first-type protein interaction domain. 8.
- the second recognition domain is specific for a target that is not recognized by the TCR recognition domain.
- the second recognition domain is specific for a target that is found on a healthy and/or non-target cell and not on a diseased and/or target cell.
- the TCR recognition domain comprises a MHC (Major Histocompatibility Complex); a MHC-peptide complex; or a MHC- peptide fusion. 12.
- composition of paragraph 11 wherein the peptide is a human or non-human peptide.
- MHC Minor Histocompatibility Antigen
- the protein interaction domains are found on an extracellular portion of the respective polypeptides. 16. The composition of any of the preceding paragraphs, a.
- protein interaction domain(s) is a leucine zipper, or any binding pair of protein interaction domains are collectively a pair of leucine zippers; b. wherein the protein interaction domain(s) is a BZip (RR) and/or a AZip (EE), or any binding pair of protein interaction domains are collectively a BZip (RR) and a AZip (EE); c. wherein the protein interaction domain(s) is a PSD95-Dlgl-zo-1 (PDZ) domain; d.
- PDZ PSD95-Dlgl-zo-1
- the protein interaction domain(s) is a streptavidin and/or a streptavidin binding protein (SBP) or any binding pair of protein interaction domains are collectively a streptavidin and a streptavidin binding protein (SBP); e. wherein the protein interaction domain(s) is a FKBP-binding domain of mTOR (FRB) and/or a FK506 binding protein (FKBP) or any binding pair of protein interaction domains are collectively a FKBP-binding domain of mTOR (FRB) and a FK506 binding protein (FKBP); f.
- SBP streptavidin binding protein
- FKBP FK506 binding protein
- the protein interaction domain(s) is a cyclophilin-Fas fusion protein (CyP- Fas) and/or a FK506 binding protein (FKBP) or any binding pair of protein interaction domains are collectively a cyclophilin-Fas fusion protein (CyP-Fas) and a FK506 binding protein (FKBP);
- the protein interaction domain(s) is a calcineurinA (CNA) and/or a FK506 binding protein (FKBP) or any binding pair of protein interaction domains are collectively a calcineurinA (CNA) and a FK506 binding protein (FKBP); h.
- the protein interaction domain(s) is a gibberellin insensitive (GIA) and/or a gibberellin insensitive dwarf1 (GID1) or any binding pair of protein interaction domains are collectively a gibberellin insensitive (GIA) and a gibberellin insensitive dwarf1 (GID1); i. wherein the protein interaction domain(s) is a Snap-tag and/or a Halo tag, or any binding pair of protein interaction domains are collectively a Snap-tag and a Halo tag; j.
- protein interaction domain(s) is a T14-3-3-cdeltaC and/or a C-Terminal peptides of PMA2 (CT52), or any binding pair of protein interaction domains are collectively a T14-3-3-cdeltaC and a C-Terminal peptides of PMA2 (CT52); k. wherein the protein interaction domain(s) is a PYL and/or a ABI, or any binding pair of protein interaction domains are collectively a PYL and a ABI; and/or l.
- protein interaction domain(s) is a nucleotide tag and/or a zinc finger domain, or any binding pair of protein interaction domains are collectively a nucleotide tag and a zinc finger domain. 17.
- the nucleotide tag is a DNA tag or dsDNA tag. 18.
- the intracellular signaling domain is a signaling domain from a protein selected from the group consisting of: TCRC; FcRy; FcRp; CD3y; CD35; CD3s; CD3C; CD22; CD79a; CD79b; CD66d; CARD11; CD2; CD7; CD27; CD28; CD30; CD40; CD54 (ICAM); CD83; CD134 (OX40); CD137 (4-1BB); CD150 (SLAMF1); CD152 (CTLA4); CD223 (LAG3); CD270 (HVEM); CD273 (PD-L2); CD274 (PD- Ll); CD278 (ICOS); DAP10; LAT; KD2C SLP76; TRIM; ZAP70; and 41BB.
- TCRC TCRC
- FcRy FcRp
- CD3y CD35
- CD3s CD3C
- CD22 CD79a; CD79b
- CD66d CARD11; CD2; CD7; CD
- a cell comprising and/or expressing the composition of any of the preceding paragraphs.
- 20 The cell of paragraph 19, wherein the TCR recognition domain comprises a MHC allogenic to the cell.
- 21 The cell of paragraph 19, wherein the TCR recognition domain comprises a peptide allogenic to the cell.
- 22 The cell of any of paragraphs 19-21, wherein the cell is a dendritic cell, regulatory T cell, or effector T cell.
- 23 The cell of any of paragraphs 19-22, wherein the cell is engineered to express the polypeptide(s) of the composition.
- 24 The cell of any of paragraphs 19-22, wherein the cell is engineered to express the signaling polypeptide of the composition. 25.
- a chimeric antigen receptor (CAR) comprising: a. an anti-CD127 and/or anti-CD45RO recognition domain; b. an intracellular signaling domain.
- a composition comprising: a first polypeptide comprising: a. an anti-CD127 and/or anti-CD45RO recognition domain; b. a first-type protein interaction domain; and a second polypeptide comprising: c. a second-type protein interaction domain; and d. an intracellular signaling domain; wherein the first-type protein interaction domain and the second-type protein interaction domain bind specifically to each other.
- a composition comprising: a first polypeptide comprising: a. an anti-CD127 recognition domain; b. a first-type protein interaction domain; a second polypeptide comprising: c. an anti-CD45RO recognition domain; d. a fifth-type protein interaction domain; and a third polypeptide comprising: e. a second-type and a fourth-type protein interaction domain; and f. an intracellular signaling domain; wherein the first-type protein interaction domain and the second-type protein interaction domain bind specifically to each other; and wherein the fourth-type protein interaction domain and the fifth-type protein interaction domain bind specifically to each other. 29. A cell comprising the CAR or composition of any of paragraphs 26-28. 30.
- the TCR recognition domain comprises a MHC allogenic to the subject.
- the TCR recognition domain comprises a MHC autologous to the transplant cells.
- the TCR recognition domain comprises a peptide allogenic to the subject.
- the TCR recognition domain comprises a peptide autologous to the transplant cells.
- the transplant is vascularized composite allotransplantation (VCA). 36.
- a composition comprising: a. a TCR recognition domain; and one or both of: b. an intracellular signaling domain; and c. a first-type biomolecular interaction domain.
- a composition comprising: a. a first polypeptide comprising at least a portion of a TCR recognition domain and a first-type biomolecular interaction domain; and b.
- a composition comprising: a. a first polypeptide comprising at least a portion of a TCR recognition domain and a first-type biomolecular interaction domain; and b. a recognition polypeptide comprising a second recognition domain and a third-type biomolecular interaction domain; wherein the first-type and third-type biomolecular interaction domains bind specifically to each other.
- a composition comprising: a.
- a first polypeptide comprising at least a portion of a TCR recognition domain and a first-type biomolecular interaction domain
- a signaling polypeptide comprising a second-type biomolecular interaction domain and an intracellular signaling domain
- a recognition polypeptide comprising a second recognition domain and a third-type biomolecular interaction domain; wherein the second-type and third-type biomolecular interaction domains compete for binding to the first-type biomolecular interaction domain.
- a composition comprising: a. a first polypeptide comprising at least a portion of a TCR recognition domain and a first-type biomolecular interaction domain; b. a signaling polypeptide comprising a second-type biomolecular interaction domain, a fourth-type biomolecular interaction domain, and an intracellular signaling domain; and c. a recognition polypeptide comprising a second recognition domain and a fifth-type biomolecular interaction domain; wherein the first-type biomolecular interaction domain and the second-type biomolecular interaction domain bind specifically to each other; and wherein the fourth-type biomolecular interaction domain and the fifth-type biomolecular interaction domain bind specifically to each other. 7.
- composition of paragraph 6 wherein the fourth-type biomolecular interaction domain and fifth-type biomolecular interaction domain have a weaker affinity than the second-type biomolecular interaction domain and first-type protein interaction domain.
- the first polypeptide further comprises a sixth-type biomolecular interaction domain and the recognition polypeptide further comprises a seventh-type biomolecular interaction domain which bind specifically to each other.
- the first polypeptide comprises the entire TCR recognition domain.
- composition of any of paragraphs 2-8 wherein the TCR recognition domain comprises at least two separate polypeptide sequences, the first polypeptide comprises at least one of the separate polypeptide sequences of the TCR recognition domain, and the first polypeptide is bound to or complexed with a second or further polypeptide sequences of the TCR recognition domain to form a TCR recognition domain.
- the TCR recognition domain comprises a non-polypeptide component.
- the second recognition domain is specific for a target that is not recognized by the TCR recognition domain.
- TCR recognition domain comprises a MHC (Major Histocompatibility Complex); a MHC-peptide complex; featureless peptide MHC; or a MHC-peptide fusion.
- MHC Major Histocompatibility Complex
- MHC-peptide complex featureless peptide MHC
- MHC-peptide fusion or a MHC-peptide fusion.
- the peptide is a Minor Histocompatibility Antigen (MiHA). 17.
- MHC-peptide complex is a monomer, dimer, trimer, tetramer, pentamer, dextramer or other oligomer form.
- MHC-peptide fusion is a monomer, dimer, trimer, tetramer, pentamer, dextramer or other oligomer form.
- composition of any of paragraphs 14-17, wherein the MHC is a dimer, trimer, tetramer, pentamer, dextramer or other oligomer form.
- MHC-peptide complex is a dimer, trimer, tetramer, pentamer, dextramer or other oligomer form.
- MHC-peptide fusion is a dimer, trimer, tetramer, pentamer, dextramer or other oligomer form.
- the composition of any of paragraphs 14-22, wherein the MHC is a MHC class I or a MHC class II. 24.
- biomolecular interaction domain(s) is a BZip (RR) and/or a AZip (EE), or any binding pair of biomolecular interaction domains are collectively a BZip (RR) and a AZip (EE); c. wherein the biomolecular interaction domain(s) is a PSD95-Dlgl-zo-1 (PDZ) domain; d. wherein the biomolecular interaction domain(s) is a streptavidin and/or a streptavidin binding biomolecular (SBP) or any binding pair of biomolecular interaction domains are collectively a streptavidin and a streptavidin binding biomolecular (SBP); e.
- PDZ PSD95-Dlgl-zo-1
- biomolecular interaction domain(s) is a FKBP-binding domain of mTOR (FRB) and/or a FK506 binding biomolecular (FKBP) or any binding pair of biomolecular interaction domains are collectively a FKBP-binding domain of mTOR (FRB) and a FK506 binding biomolecular (FKBP); f.
- biomolecular interaction domain(s) is a cyclophilin-Fas fusion biomolecular (CyP-Fas) and/or a FK506 binding biomolecular (FKBP) or any binding pair of biomolecular interaction domains are collectively a cyclophilin-Fas fusion biomolecular (CyP-Fas) and a FK506 binding biomolecular (FKBP); g.
- biomolecular interaction domain(s) is a calcineurin A (CNA) and/or a FK506 binding biomolecular (FKBP) or any binding pair of biomolecular interaction domains are collectively a calcineurin A (CNA) and a FK506 binding biomolecular (FKBP); h. wherein the biomolecular interaction domain(s) is a gibberellin insensitive (GIA) and/or a gibberellin insensitive dwarf1 (GID1) or any binding pair of biomolecular interaction domains are collectively a gibberellin insensitive (GIA) and a gibberellin insensitive dwarf1 (GID1); i.
- biomolecular interaction domain(s) is a Snap-tag and/or a Halo tag, or any binding pair of biomolecular interaction domains are collectively a Snap-tag and a Halo tag; j. wherein the biomolecular interaction domain(s) is a T14-3-3-cdeltaC and/or a C- Terminal peptides of PMA2 (CT52), or any binding pair of biomolecular interaction domains are collectively a T14-3-3-cdeltaC and a C-Terminal peptides of PMA2 (CT52); k.
- biomolecular interaction domain(s) is a PYL and/or a ABI, or any binding pair of biomolecular interaction domains are collectively a PYL and a ABI; l. wherein the biomolecular interaction domain(s) is a nucleotide tag and/or a zinc finger domain, or any binding pair of biomolecular interaction domains are collectively a nucleotide tag and a zinc finger domain; m. wherein the biomolecular interaction domain(s) is a nucleotide tag, or any binding pair of biomolecular interaction domains are collectively a pair of nucleotide tags; n.
- biomolecular interaction domain(s) is a Fluorescein isothiocyanate (FITC) and/or a FITC binding biomolecular or any binding pair of protein interaction domains are collectively a FITC and a FITC binding protein; and/or o.
- the protein interaction domain(s) is a (R)-Phycoerythrin (R-PE/ PE) and/or a R-PE/PE binding protein or any binding pair of protein interaction domains are collectively a (R)-Phycoerythrin (R-PE/ PE) and a R-PE/PE binding protein.
- the intracellular signaling domain comprises or is a signaling domain from one or more proteins selected from the group consisting of: TCR ⁇ , FcR ⁇ , FcR ⁇ , CD3 ⁇ ; CD35; CD3 ⁇ ; CD3C; CD22; CD79a; CD79b; CD66d; CARD11; CD2; CD7; CD27; CD28; CD30; CD40; CD54 (ICAM); CD83; CD134 (OX40); CD137 (4-1BB); CD150 (SLAMF1); CD152 (CTLA4); CD223 (LAG3); CD270 (HVEM); CD273 (PD-L2); CD274 (PD- Ll); CD278 (ICOS); DAP10; LAT; KD2C SLP76; TRIM; and ZAP70.
- proteins selected from the group consisting of: TCR ⁇ , FcR ⁇ , FcR ⁇ , CD3 ⁇ ; CD35; CD3 ⁇ ; CD3C; CD22; CD79a; CD79b; CD66d; CARD11; CD
- a cell comprising and/or expressing the composition of any of the preceding paragraphs.
- 31. A composition comprising a first polypeptide of any of the preceding paragraphs and a cell expressing or comprising the signaling polypeptide of any of the preceding paragraphs.
- 32. The cell or composition of any of paragraphs 30-31, wherein the TCR recognition domain comprises a MHC allogeneic, autologous, or xenogeneic to the cell.
- 33. The cell or composition of any of paragraphs 30-32, wherein the TCR recognition domain comprises a synthetic MHC. 34.
- a chimeric antigen receptor comprising: a. an anti-CD127 and/or anti-CD45RO recognition domain; b. an intracellular signaling domain.
- a composition comprising: a first polypeptide comprising: a. an anti-CD127 and/or anti-CD45RO recognition domain; b.
- a composition comprising: a first polypeptide comprising: a. an anti-CD127 recognition domain; b. a first-type protein interaction domain; a second polypeptide comprising: c. an anti-CD45RO recognition domain; d. a fifth-type protein interaction domain; and a third polypeptide comprising: e. a second-type and a fourth-type protein interaction domain; and f.
- a cell comprising the CAR or composition of any of paragraphs 41-43.
- the TCR recognition domain comprises a MHC autologous to the transplant cells.
- the TCR recognition domain comprises a MHC xenogeneic to the transplant cells.
- the TCR recognition domain comprises a MHC and a peptide, wherein the peptide is allogeneic to the subject.
- the TCR recognition domain comprises a MHC and a peptide, wherein the peptide is autologous to the transplant cells.
- the TCR recognition domain comprises a MHC and a peptide, wherein the peptide is autologous to the transplant cells. 52.
- autoimmune disease is type 1 diabetes, vitiligo, multiple sclerosis, alopecia, celiac disease, pemphigus, rheumatoid arthritis, or scleroderma. 57.
- autoimmune disease is thyroiditis, type 1 diabetes mellitus, Hashimoto's thyroiditis, Graves' disease, celiac disease, multiple sclerosis, Guillain-Barre syndrome, Addison's disease, and Raynaud's phenomenon, Goodpasture's disease, arthritis (rheumatoid arthritis such as acute arthritis, chronic rheumatoid arthritis, gout or gouty arthritis, acute gouty arthritis, acute immunological arthritis, chronic inflammatory arthritis, degenerative arthritis, type II collagen-induced arthritis, infectious arthritis, Lyme arthritis (e.g., post treatment Lyme disease syndrome), proliferative arthritis, psoriatic arthritis, Still's disease, vertebral arthritis, and juvenile-onset rheumatoid arthritis, arthritis chronica progrediente, arthritis deformans, polyarthritis chronica primaria, reactive arthritis, and ankylosing spondylitis), palindromic arthritis, inflammatory hyperosis, rheumatoid arthritis
- T cell mediated immune response is an anti-drug specific response to a biologic, cell therapy, and/or gene therapy.
- the biologic, cell-therapy, or gene therapy is an adeno- associated virus (AAV) gene therapy, a genome editing agent, or enzyme replacement therapy. 60.
- AAV adeno- associated virus
- the disease is type 1 diabetes and the TCR recognition domain comprises sequences with at least 80% or at least 95% sequence identity to: one or more of SEQ ID NOs: 8-17; HLA-A*0201 and at least one of SEQ ID NOs: 2013-2016 and 2031- 2033; or HLA-A*02:01 and at least one of SEQ ID NOs: 20128-2129. 61.
- the disease is vitiligo and the TCR recognition domain comprises sequences with at least 80% or at least 95% sequence identity to: SEQ ID NO: 18, 19, and one of 20-22; or comprises HLA-A*0201 and SEQ ID NO: 2018; or HLA-A*0301 and SEQ ID NO: 2019; or comprises HLA-A*2402 and SEQ ID NO: 2020; or HLA-A*0101 and SEQ ID NO: 2021. 62.
- the method is a method of treating and/or preventing GvHD and the TCR recognition domain comprises sequences with at least 80% or at least 95% sequence identity to: HLA-A*0101 and at least one of SEQ ID NOs: 2034-2037; or HLA-B*0702 and SEQ ID NO: 2038; or HLA-B*0801 and SEQ ID NO: 2039. 63.
- the disease is type 1 diabetes and the TCR recognition domain comprises one or more of SEQ ID NOs: 8-17; comprises HLA-A*0201 and at least one of SEQ ID NOs: 2013-2016 and 2031-2033; or comprises HLA-A*02:01 and at least one of SEQ ID NOs: 20128-2129. 64.
- the disease is vitiligo and the TCR recognition domain comprises SEQ ID NO: 18, 19, and one of 20-22; or comprises HLA-A*0201 and SEQ ID NO: 2018; or comprises HLA-A*0301 and SEQ ID NO: 2019; or comprises HLA-A*2402 and SEQ ID NO: 2020; or comprises HLA-A*0101 and SEQ ID NO: 2021. 65.
- a composition comprising: a. a TCR recognition domain; and one or both of: b. an intracellular signaling domain; and c. a biomolecular interaction domain. 2.
- composition of paragraph 1 comprising a TCR recognition domain and a biomolecular interaction domain. 3.
- the biomolecular interaction domain of c) is a first- type biomolecular interaction domain and the composition further comprises a signaling polypeptide comprising a second-type biomolecular interaction domain and an intracellular signaling domain; wherein the first-type and second-type biomolecular interaction domains bind specifically to each other.
- the TCR recognition domain comprises a MHC (Major Histocompatibility Complex); a MHC-peptide complex; a featureless peptide MHC; or a MHC-peptide fusion.
- the TCR recognition domain comprises a CD1 domain or a CD1 domain-ligand complex or fusion. 7. The composition of paragraph 6, wherein the CD1 is CD1d. 8. The composition of paragraph 1, wherein the TCR recognition domain is a monomer, dimer, trimer, tetramer, pentamer, dextramer or other oligomer form. 9. The composition of paragraph 1, a. wherein the biomolecular interaction domain(s) is a leucine zipper, or any binding pair of biomolecular interaction domains are collectively a pair of leucine zippers; b.
- biomolecular interaction domain(s) is a BZip (RR) and/or a AZip (EE), or any binding pair of biomolecular interaction domains are collectively a BZip (RR) and a AZip (EE); c. wherein the biomolecular interaction domain(s) is a PSD95-Dlgl-zo-1 (PDZ) domain; d. wherein the biomolecular interaction domain(s) is a streptavidin and/or a streptavidin binding biomolecular (SBP) or any binding pair of biomolecular interaction domains are collectively a streptavidin and a streptavidin binding biomolecular (SBP); e.
- PDZ PSD95-Dlgl-zo-1
- biomolecular interaction domain(s) is a FKBP-binding domain of mTOR (FRB) and/or a FK506 binding biomolecular (FKBP) or any binding pair of biomolecular interaction domains are collectively a FKBP-binding domain of mTOR (FRB) and a FK506 binding biomolecular (FKBP); f.
- biomolecular interaction domain(s) is a cyclophilin-Fas fusion biomolecular (CyP-Fas) and/or a FK506 binding biomolecular (FKBP) or any binding pair of biomolecular interaction domains are collectively a cyclophilin-Fas fusion biomolecular (CyP-Fas) and a FK506 binding biomolecular (FKBP); g.
- biomolecular interaction domain(s) is a calcineurin A (CNA) and/or a FK506 binding biomolecular (FKBP) or any binding pair of biomolecular interaction domains are collectively a calcineurin A (CNA) and a FK506 binding biomolecular (FKBP); h. wherein the biomolecular interaction domain(s) is a gibberellin insensitive (GIA) and/or a gibberellin insensitive dwarf1 (GID1) or any binding pair of biomolecular interaction domains are collectively a gibberellin insensitive (GIA) and a gibberellin insensitive dwarf1 (GID1); i.
- biomolecular interaction domain(s) is a Snap-tag and/or a Halo tag, or any binding pair of biomolecular interaction domains are collectively a Snap-tag and a Halo tag; j. wherein the biomolecular interaction domain(s) is a T14-3-3-cdeltaC and/or a C- Terminal peptides of PMA2 (CT52), or any binding pair of biomolecular interaction domains are collectively a T14-3-3-cdeltaC and a C-Terminal peptides of PMA2 (CT52); k.
- biomolecular interaction domain(s) is a PYL and/or a ABI, or any binding pair of biomolecular interaction domains are collectively a PYL and a ABI; l. wherein the biomolecular interaction domain(s) is a nucleotide tag and/or a zinc finger domain, or any binding pair of biomolecular interaction domains are collectively a nucleotide tag and a zinc finger domain; m. wherein the biomolecular interaction domain(s) is a nucleotide tag, or any binding pair of biomolecular interaction domains are collectively a pair of nucleotide tags; n.
- biomolecular interaction domain(s) is a Fluorescein isothiocyanate (FITC) and/or a FITC binding biomolecular or any binding pair of protein interaction domains are collectively a FITC and a FITC binding protein; and/or o.
- the protein interaction domain(s) is a (R)-Phycoerythrin (R-PE/ PE) and/or a R-PE/PE binding protein or any binding pair of protein interaction domains are collectively a (R)-Phycoerythrin (R-PE/ PE) and a R-PE/PE binding protein.
- the nucleotide tag is a DNA tag or dsDNA tag.
- composition of paragraph 2 further comprising a cell expressing or comprising the signaling polypeptide.
- TCR recognition domain is allogeneic, autologous, or xenogenic to the cell.
- TCR recognition domain is synthetic.
- the TCR recognition domain comprises a MHC and a peptide, wherein the peptide is allogeneic, autologous, or xenogenic to the cell.
- the TCR recognition domain comprises a MHC and a peptide, wherein the peptide is synthetic.
- the cell is a NK cell, dendritic cell, regulatory T cell, or effector T cell. 17.
- composition of paragraph 11 wherein the cell is further engineered to knockdown the native MHCI/II expressed on the cell surface.
- the transplant is an allogeneic hematopoietic stem cell or solid organ transplantation.
- the malignant T cell condition is T cell acute lymphoblastic leukemia or T cell lymphoblastic lymphoma. 21.
- the autoimmune disease is type 1 diabetes, vitiligo, multiple sclerosis, alopecia, celiac disease, pemphigus, rheumatoid arthritis, or scleroderma.
- the T cell mediated immune response is an anti-drug specific response to a biologic, cell therapy, and/or gene therapy.
- the biologic, cell-therapy, or gene therapy is an adeno- associated virus (AAV) gene therapy, a genome editing agent, or enzyme replacement therapy.
- the disease is type 1 diabetes and the TCR recognition domain comprises sequences with at least 95% sequence identity to: one or more of SEQ ID NOs: 8-17; HLA-A*0201 and at least one of SEQ ID NOs: 2013-2016 and 2031-2033; or HLA-A*02:01 and at least one of SEQ ID NOs: 20128-2129. 25.
- the disease is vitiligo and the TCR recognition domain comprises sequences with at least 95% sequence identity to: SEQ ID NO: 18, 19, and one of 20-22; or HLA-A*0201 and SEQ ID NO: 2018; or HLA-A*0301 and SEQ ID NO: 2019; or comprises HLA-A*2402 and SEQ ID NO: 2020; or HLA-A*0101 and SEQ ID NO: 2021. 26.
- the method is a method of treating and/or preventing GvHD and the TCR recognition domain comprises sequences with at least 95% sequence identity to: HLA-A*0101 and at least one of SEQ ID NOs: 2034-2037; or HLA-B*0702 and SEQ ID NO: 2038; or HLA-B*0801 and SEQ ID NO: 2039.
- EXAMPLES Example 1 Auto- and allo-reactive T cells attacking patient's or donor's cells or organs is the major cause of autoimmunity and transplant rejection. Current treatments involve stringent immunosuppressant therapy, which can lead to severe side effects.
- T cells engineered with a Chimeric antigen ligand can redirect their specificity toward the pathologic T cells.
- T cells engineered with a Chimeric antigen receptor can redirect their specificity and have already been approved to treat some types of B cell malignancies.
- engineered regulatory T cell which can inhibit immune reactions in an antigen-dependent manner, is under investigation to expand the application of CAR T cells therapies such as autoimmune disease and transplant rejection.
- This CAL and/or CAR is a composed of a peptide-HLA (e.g., a monomer or multimer or oligomer thereof) as the recognition domain fused to signaling domains from T cell receptors.
- a split version of the system can also be generated where the sytem is composed of two pieces.
- One piece is an universal CAR with T cell signaling domains as the intracellular portion and a biomolecular interaction domain as the extracellular domain.
- the second piece is an adaptor molecule, e.g, a CAL composed of a peptide-HLA oligomers (or monomer or multimer) fused to the cognate biomolecular interaction domain.
- DCs Dendritic cells
- pDCs plasmacytoid DCs
- DCs are essential mediators of pro- inflammatory or anti-inflammatory (tolerogenic) responses.
- the subsets of DCs that suppress immune responses are generally known as tolerogenic DCs because of their functions in inducing T cell apoptosis, anergy and regulatory T cells (Tregs).
- Tregs regulatory T cells
- a tolerogenic state in DCs can be induced using several pharmacological agents, such as cyclosporine A, rapamycin, dexamethasone, vitamin A, vitamin D or other cytokines and growth factors. Recently, the insertion of exogenous DNA to enhance tol-DC function has been investigated as a therapeutic possibility for treating autoimmune diseases.
- the effect of these DCs are not specific.
- Alloreactive immune cells can be suppressed in a specific manner using the methods described herein, e.g., by using a universal UniCAR DC system that presents the donors' pMHC tetra / dextramers (e.g., the CAL) and binds to a genetically engineered DC (K/O MHC I / II but presents donor peptide on the tetra /dextramer), .
- the pMHC can be provided as a monomer, oligomer, or multimer.
- Any form of the recipient pMHC (e.g., monomer / oligomer / tetramer / dextramer)+ targeting module (e.g, the CAL) can be used with CAR-DC for the suppression of allo-reactive T- cells in the recipients.
- a library of identified different alloantigens + correlated or associated with MHC monomers/ oligomers / tetramers / dextramers (e.g. HLA-A2+insulin peptide) and attached to the targeting molecules of the CAL and/or CAR system can be produced.
- Such a library can be commercially available to be combined with DCs for targeting and destroying the auto reactive T- cells.
- Cells can be produced that are genetically modified with the specific gene deletion of some genes (IL-12 and NF- ⁇ B, MHC I and MHC II [The targeting module will provide these cells with the donor's MHCs]) and insertion of some other genes (IL-10, TGF- ⁇ , CTLA-4 and SOCS1). These cells can be commercially ready to use and could be combined with the specific donor, pMHC and targeting module chosen from the aforementioned library.
- Alloreactive memory T cells have been shown to have the key role in the activation of the recipient immune system and rejection of the allograft. They are known to be the main barrier for tolerance induction through mixed chimerism and other strategies.
- compositions of the invention can be used to decrease or eliminate the need for improved immunosuppressant therapy in the context of transplantation.
- the engraftment of hematopoietic stem cells can achieve durable mixed chimerism with minimal or no need for toxic preconditioning protocols.
- donor pMHC monomer/oligomer/tetramer/dextramer; e.g., a CAL
- CAR T effector for the abrogation of the allo/xeno reactive T-cells in the recipient
- a library of different Donor MHC monomers/oligomers/tetramers attached to the targeting molecules (e.g., the CALs) of the SUPRA / UNI / universal CAL and/or CAR system can be produced to be commercially available.
- T cells are critical contributors to autoimmune diseases.
- Conventional T (Teff or T helper) cell subsets that play a role in B cell activation and differentiation produce various inflammatory cytokines and destroy target cells with direct cytotoxicity.
- CAR-T cells have been used to destroy autoimmune B and T cells in a fashion similar to the way in which CD19CAR T cells target and destroy leukemia cells.
- autoreactive memory T and B-cells Targeting autoreactive memory T and B-cells has shown some results. As these methods are nonspecific, they induce some extent of generalized immune suppression and they are not completely effective. The correlation of particular peptide+MHC molecules with certain autoimmune conditions have been expansively studied. By using the available human/animal model pMHC monomer/oligomer/tetra/dextramers and combining them with the SUPRA / UNI / universal CAL and/or CAR technology, we are able to target the autoreactive immune cells in a highly specific manner.
- any form of auto-antigen on pMHC (monomer/oligomer/tetramer/dextramer) (e.g., the CAL) is used in combination with CAR T effector / Treg to abrogate autoreactive T-cells in the recipient, the need for immunomodulatory drugs would be decreased or eliminated.
- a library of identified autoantigens and correlated MHC monomer/oligomer/tetra/dextramers e.g.
- HLA- A2+insulin peptide) attached to the targeting molecules of the SUPRA / UNI / universal CAL and/or CAR system can be produced to be commercially available to be combined with ideal SUPRA or universal CAR T-reg / T effector for targeting and destroying the autoreactive T-cells.
- Example 5 Although new advances have increased survival after allogeneic hematopoietic stem cell transplantation (HCT), chronic graft-versus-host disease (GvHD) is still the leading cause of late morbidity and mortality after transplant. Current treatment choices are limited in efficacy specifically in steroid-refractory disease, and there is no robust data to help with management decisions.
- Adoptive T cell therapy refers to the therapeutic use of T cells.
- T cells genetically engineered to express chimeric antigen receptors (CAR) constitute the most clinically advanced form of ACT approved to date for the treatment of CD19-positive leukemias and lymphomas and have produced remarkable results in the clinic.
- CAR chimeric antigen receptors
- the technology described herein permits the opportunity to target diseased cells with specific antigens or receptors very accurately.
- GvHD universal UNICAR T cells can be designed to find the recipient reactive T cell in the donor T cell populations.
- the pMHC of the recipients is fused to a Target Module that binds to the UniCAR (e.g, to form a CAL)
- this system can recognize the TCR repertoires in the donor T/B cell population that can bind to those MHCs.
- the recipients’ pMHCs are fused to a target module that binds to the universal CAR, this system can recognize and bind to the T cell repertoires in the donor T cell population that can bind to those MHCs.
- the recipient pMHC (monomer / oligomer / tetramer / dextramer) (e.g., a CAL) is used in combination with CAR T effector / Treg for the abrogation of the recipient reactive T-cells in the donor HSC, these reactive T-cells will be depleted and GvHD would not happen.
- a library of identified different antigens + correlated human / animal models MHC tetramers / dextramers e.g.
- HLA-A2+ peptide) attached to the targeting molecules of the SUPRA / UNI CAR system can be produced to be commercially being available to be combined with ideal SUPRA CAR T-reg / T effector for targeting and destroying the reactive T-cells against the recipients.
- a library of wild type and/or synthetic MHC monomers/oligomers (e.g. HLA-A2+ peptide) attached to the targeting molecules of the universal CAL system can be generated to be mixed with universal CAL T-reg / T effector for targeting and killing the reactive T-cells against the recipients within the donor T cell population.
- Example 6 Although new advances have increased survival after allogeneic hematopoietic stem cell transplantation (HCT), chronic graft-versus-host disease (GvHD) is still the leading cause of late morbidity and mortality after transplant. Current treatment choices are limited in efficacy specifically in steroid-refractory disease, and there is no robust data to help with management decisions.
- Adoptive T cell therapy refers to the therapeutic use of T cells. T cells genetically engineered to express chimeric antigen receptors (CAR) constitute the most clinically advanced form of ACT approved to date for the treatment of CD19-positive leukemias and lymphomas and produced remarkable results in clinical. This technology provides the opportunity to target the cells with specific antigens or receptors very accurately.
- UNICAR T cells can be designed to find the recipient reactive T cell in the donor T cell populations. If the pMHC of the recipients be fused to a Target Module that bind to the UniCAR, this system can recognize the TCR repertoires in the donor T/B cell population that can bind to those MHCs. [00317] If the recipient pMHC (tetramer / dextramer) is used in combination with CAR T effector / Treg for the abrogation of the recipient reactive T-cells in the donor HSC, these cells will be depleted and GvHD would not happen. A library of identified different antigens + correlated human / animal models MHC tetramers / dextramers (e.g.
- HLA-A2+ peptide) attached to the targeting molecules of the SUPRA / UNI CAR system can be produced to be commercially being available to be combined with ideal SUPRA CAR T-reg / T effector for targeting and destroying the reactive T- cells against the recipients.
- Example 7 - Engineered Lymphocytes for Prevention of Pediatrics Vascularized Composite Allograft Rejection Engineered [00318] Described herein is the development of a clinically applicable tolerance-inducing regimen for VCA transplantation through the establishment of stable mixed chimerism, augmented by the state-of-the-art CAL and/or CAR T cell adoptive immunotherapy and synthetic biology.
- Aim 1 It has been shown that CD127+/CD45RO+ memory T cells have a significant role as central, effector and stem cell memory T cells and are known as the most potent constituents of the alloreactive T cell repertoire. These cells have been shown to be the major contributor in chronic rejection of the allograft.
- a CAL and/or CAR T-effector cell can be generated with scfv against two general markers of memory T cells (CD127+ and CD45RO+). This will be followed by the induction of mixed chimerism protocol in a double humanized mouse model.
- Aim 2 Due to the diversity of antigen-specific T cells in the context of transplantation, a CAL and/or CAR system is provided that has the flexibility to locate and attack different alloreactive T cells simultaneously. CAL and/or CAR T-eff cells with donor pMHC can target alloreactive T-cells that have TCR against the donor MHCs. We will start with single and double specific antigen-MHC systems.
- Example 8 Engineered Lymphocytes for Prevention of Pediatrics Vascularized Composite Allograft Rejection
- VCA vascularized composite allotransplantation
- VCA represents a unique new treatment option for severe soft tissue defects following burn injury to achieve both psychosocial and functional rehabilitation 3 .
- the shortcoming is that despite the use of potent immunosuppressive drugs, acute rejection of “foreign” VCA occurs in up to 90% of patients 5 .
- our immune system has gained the ability to recognize self from nonself cells and attack the “foreigners” by cells such as T Lymphocytes.
- HLA Human Leukocyte Antigen
- MHC Major Histocompatibility Complex
- MiHA Minor Histocompatibility Antigens
- T-cells are trained to recognize self from non-self MHC / peptides. Each person can present a combination of 12 different MHC class I / II on the surface of his/her cells. (HLA) typing is used to match recipients and donors MHCs for transplants.
- a close match between a donor’s and a recipient's HLA markers is essential for a successful transplant outcome.
- a potential donor must match a minimum of 6-8 HLA markers which makes finding an ideal donor very difficult. Even after finding the semi ideal donor these patients will be on immunosuppressants for the rest of their life 8 . While these drugs are generally effective, the sequelae of such chronic immunosuppression are well known, and most recipients continue to develop myriad side effects and complications, including opportunistic infections, multiple organ dysfunction, and malignancies.
- Transplantation tolerance that allows for the elimination of immunosuppressive drugs, has been the “holy grail” for transplantation medicine since its beginnings 9 .
- the present approach to achieving stable mixed chimerism is to utilize advanced engineering techniques to generate T cell therapeutics to specifically delete the recipient’s alloreactive memory immune cells that are reactive against the donor bone marrow and tissue cells.
- CAR chimeric antigen receptor
- First-generation CARs contained only an extracellular antigen-binding domain, a transmembrane domain, and the signaling domain of CD3z.
- intracellular costimulatory domains derived from either CD28 / 4-1BB were added to enhance proliferation, persistence, and activity 19 .
- SUPRA split, universal, and “programmable”
- mice can be conditioned with intraperitoneal mAb injections including, anti-CD8 and anti-CD40L, anti-CD154, costim. blockade (on day 0 with respect to the BMT). This will be combined with, TBI to be given 6 h before injection of 2.510 5 bone marrow cells (BMC).
- BMC bone marrow cells
- a CAL and/or CAR designed to target and destroy alloreactive memory T cells thereby improving the engraftment of hematopoietic stem cells and achieving durable mixed chimerism with minimal or no need for toxic preconditioning protocols.
- a second approach is to design CAL and/or CARs that specifically bind to alloreactive TCRs on the recipient’s T cells.
- scFvs single-chain variable fragments
- we will use peptide-MHC multimers as the ligand recognition domain on the CAL and/or CAR.
- scFvs single-chain variable fragments
- a two- component split CAL and/or CAR design composed of universal CAL and/or CAR and an adaptor molecule that bridges the CAL and/or CAR T cells to alloreactive T cells.
- the universal CAL and/or CAR comprises a scFv targeting FITC fused to TCR signaling domains.
- the adaptor molecules consist of pMHC tetramer fused to a FITC. The addition of pMHC tetramer will bind to the alloreactive TCR and recruit the universal CAL and/or CAR T cell to kill alloreactive T cells. This design allows us to be able to target many variants of TCR with only one CAL and/or CAR.
- the donor MHCs will be loaded with donor tissue specific peptides that will be generated by utilizing previously published protocols.
- Data- Targeting Alloreactive T-cells with peptide-MHC tetramers The following have been addressed by in vitro data: 1. Whether targeting a memory T cell by using CAR T + pMHC multimer is obtainable.
- the anti-FITC CAR expressed by the Jurkat cells is a CAR and the pMHC multimer is a CAL.
- A. would the Tetramer efficiently bind to the CAL and/or CAR and target T cells? B.
- Fig.1 shows tetramers attaching to and staining OTi target T cells in a dose dependent fashion.
- Nucleotides 1339-1677 CD3 ⁇ . Nucleotides 1678-2388 mCHERRY. gccgccaccATGGCCTTACCAGTGACCGCCTTGCTCCTGCCGCTGGCCTTGCTGCTCCACGCC GCCAGGCCGGCTGCAcaagttcagctcgttgaatccggcggaaaccttgttcaaccggggggttccctccgccttagttgtgccgcat ctggttttacgtttggatcatttttccatgtcatgggtgcggcaggctcccggggggggactcgaatgggttgcgggtcttagcgcccgatcaagcctc actcactatgcagatagcgtaaaggcaggtttacaatctcacgagacaacgcgaagaactcagtctatcttcagatgaactct
- Residues 1-21 CD8a leading peptide.
- Residues 22-274 anti-FITC ScFv.
- Residues 275-284 myc.
- Residues 289-333 CD8 hinge.
- Residues 334-401 CD28.
- Residues 402-443 4-1BB.
- Residues 444-556 CD3 ⁇ . Residues 557-792 mCHERRY.
- Primary CD8 anti-FITC CAR T cells can specifically target and kill alloreactive OTi cells.
- the previous set of experiments with Jurkat +adaptor +OTi target cells confirmed the feasibility of targeting alloreactive T cells by adaptor molecules and showed activation of CAR T cells against the target cells.
- primary CD8 CAR T killer cells + adaptor molecules can be used not only to identify but also to kill the OTi T cells.
- the binding between pMHC and target cell was not causing activation of target T cells in contrary to the adaptor (FITC+ pMHC, e.g., the CAL) - CAR T cells binding.
- pMHC multimers for specifically targeting and destroying allo / autoreactive T cells responsible for graft rejection and autoimmune diseases.
- Our approach is the first demonstration of utilizing pMHC multimer (e.g, CAL) in combination with CAR T cell technology for specific targeting of alloreactive T cell population.
- pMHC multimer e.g, CAL
- CAR T cell technology for specific targeting of alloreactive T cell population.
- This approach is the most specific strategy for eliminating the destructive role of alloreactive T cells in transplantation.
- the potential application of this method extends far beyond elimination of immunosuppressants in transplantation rejection and GVHD treatment and could be applicable to many autoimmune conditions including type 1 diabetes, multiple sclerosis, rheumatoid arthritis, scleroderma, and myriad disorders that originate from auto reactive T cells dysfunction.
- [00348] Provided herein is a novel approach for targeting the memory repertoire with anti CD45RO / CD127 with CAL and/or CAR T cell therapy. Although this strategy might not be as specific as using pMHC multimers, it still conveys a significant novelty. By depleting the memory T cells population there is much less of a need for induction protocols. [00349] Also contemplated herein are individualized peptide libraries based on the differences between the recipient and the donor proteome and immune-peptidome. Hundreds to thousands of MHC-associated peptides can now be identified in a single measurement using optimal biological model systems. As mentioned earlier, studies involving MHC identical grafts indicate that minor histocompatibility antigens may also mediate rejection.
- HIPP Human Immuno-Peptidome Project
- the donor multimers MHCs can be generated using commercially available services, unloaded with UV and then loaded with donor tissue specific peptides that will be generated by utilizing previously published protocols. By utilizing these peptides in combination with exchangeable MHC tetramer systems, we can employ these peptides to identify and target donor reactive T cells. [00351] As described herein, selective depletion of memory T cells with CAR-T cell therapy can help in achieving durable mixed chimerism and tolerance which lead to the elimination or decrease in immunosuppressive regimens. Currently this challenge is addressed by administering intense whole- body irradiation with several immunosuppressant combinations for targeting all the immune cell populations.
- This (SUPRA) CAR is a two-component receptor system comprises a universal receptor (zipCAR) expressed on T cells and a targeting scFv adaptor (zipFv).
- the zipCAR universal receptor is generated from the fusion of intracellular signaling domains ( Figure 6) and a leucine zipper as the extracellular domain.
- the zipFv adaptor molecule is generated from the fusion of a cognate leucine zipper and an scFv.
- the scFv on the zipFv binds to the antigen, while the leucine zipper binds to and activates the zipCAL and/or zipCAR on the T cells.
- these features can mitigate over-activation and enhance specificity.
- the SUPRA CAL and/or CAR system can also be designed to perform AND combinatorial logic of antigen recognition.
- Orthogonal leucine zipper pairs can be used to generate CAL and/or CARs with split signaling domains (e.g., CD3z, CD28, 4-1BB), thus enabling independent and simultaneous control of these pathways (Fig 6B).
- T cells need both CD3z and costimulatory signaling simultaneously to be fully activated therefore, each CAL and/or CAR can be readily paired with scFvs that target different antigens, thereby enabling two antigen combinatorial and logical antigen sensing.
- SUPRA CAR T cells can also logically respond to multiple antigens for improved target specificity.
- orthogonal SUPRA CARs can be used to inducibly regulate multiple signaling pathways or T cell subtypes to increase the breadth of immune responses that can be achieved.
- zipCAR Receptor Construct Design As described herein, AND logic CAL and/or CAR T are generated by fusing different leucine zippers to the hinge region of the human CD8a chain and transmembrane and cytoplasmic regions of the human CD28, or CD3z signaling endo-domains. All CAL and/or CARs can contain a myc tag to verify surface expression. Besides, these primary T cells will also be fused to mCherry after CD3z chain to visualize expression.
- zipFv Construct Design The zipFv molecules contain an scFv against CD127 fused to an SYN2 (to stimulate the CD3z SYN1 zipCAR), and an scFv against CD45RO fused to a JUN zipper (to stimulate the costimulatory FOS zipCAR).
- the scfvs sequences are available and can be constructed rapidly through commercial DNA synthesis 28 .
- zipCAR transduction Human PBMC are purified using separation kits. are introduced into primary human T cells via retroviral transduction. Expression of zipCARs is quantified via myc and V5-tag immunostaining and flow cytometry. As controls, a zipCAR that contains both CD3z and CD28 (i.e.
- SYN1-CD28-CD3z can be used.
- This zipCAR cannot perform logic computation.
- a CD45RO-SYN2 or CD127-SYN2 zipFv can be added to ensure that zipCARs can kill CD45RO+ or CD127+ cells.
- These controls will also serve as reference for specificity.
- [00359] Evaluate the activity and efficacy of anti-CD127+/CD45RO+CAR T cells in vitro [00360] Co-culturing PBMCs with CD127+ /CD45RO+ cells with SUPRA CAR T cells to investigate the specificity of targeting [00361] This experiment examines whether this system specifically kills CD127+ and CD45RO+ double positive T cells amongst a large population of PBMCs.
- PBMCs are obtained either from known VCA recipients or Mass General Blood Bank Center after being approved by IRB.
- human CD45RO+ and CD127+ are stained to identify the initial percentage of memory cells in the whole population (CD45RO /CD127 double positive vs double neg).
- PBMCs are co-cultured with zipCAR expressing CD8+ T cells with zipFvs (anti CD127 and anti CD45RO) with the same condition.
- zipCAR expressing CD8+ T cells with zipFvs (anti CD127 and anti CD45RO) with the same condition.
- engineered T cells and zipFvs will be cocultured with the memory T cells at 3 different zipFvs concentrations (5, 25, 50ng/well) to determine the correlation between cell killing and zipFv concentration.
- T cell activation CD69 expression, IL-2 and IFN-g in the media
- cytotoxicity against memory T cells no. of remaining live cells
- the proliferation of T cells will be measured by cell counting. All conditions will be tested at least in triplicate.
- PBMCs hold an alloreactive population of memory T cells (CD45RO+/ CD127+) and makes this a reliable model for studying their depletion effects on the rejection of the graft.
- the number of mice used per group in this study is chosen based on the recommendations from previous studies 29 .
- irradiated NSG (MHC I/II K/O) (3 Gy) mice will be injected with 5 ⁇ 105PBMC cells and human peripheral reconstitution is measured by flow cytometry every other week 30 .
- NSG MHC I/II K/O mouse model that was generated by using the VCA-recipient PBMCs are used as the foundation for induction of mixed chimerism.
- NSG Hu-PBMCs are treated with / without (ctrl) anti CD45RO/CD127 CAR T to deplete the memory compartment of their T cells population (Figs.8A, 8B).
- Fetal CD34+ HSC can be obtained from Advanced Bioscience Resources. Fetal HSCs are utilized to evaluate the generation and establishment of multi-lineage engraftment by using previously published Mixed Chimerism Induction Protocols (MCIP) in mice 23. In short, male mice are treated with anti CD154, anti-CD40, CTLA-4 and receive 3 Gy TBI. Six hours later they receive 2 ⁇ 105CD34+ HSC bone marrow cells. Engraftment is determined by measuring the presence of donor myeloid and lymphoid cells at different time points. Mixed chimerism is defined as at least 5% of leukocytes in each of the lineages being donor derived.
- MCIP Mixed Chimerism Induction Protocols
- the aforementioned humanized mouse model (VCA-Recipient PBMCs-CD34 HSC) is utilized as a skin graft model to test the effect of depleting CD45RO / CD127 cells on allograft survival.
- Fetal skin graft is obtained from Advanced Bioscience Resources from the same CD34+ HSC donor.
- skin grafts from the HSC donor and another allogeneic source are used to test whether nonspecific depletion of memory T cell repertoire affects the skin grafts survival (Fig.8, 9). Mice are followed until graft rejection or POD 100.
- CD45RO is the marker that is mainly expressed on the T cells but CD127 is a more common surface marker and can be found on B and dendritic cells.
- CD45RO+ population can be targeted, which consists of not only central memory T cells but also targets effector T cells as well. This will cause more T cells to be targeted nonspecifically but simultaneously might decrease the chance of immune reaction against the graft.
- Some of the CAR T effector cells will express memory phenotype such as CD45RO /CD127 after remaining activated for a period of time.
- Monoclonal anti-donor HLA antibody is used for monitoring of chimerism in the various leukocyte lineages, including CD3+/CD8+, CD3+/CD4+, CD3+/CD8-/CD4- (naive T cells), T-Regs, naive and mature B cells (CD19+/CD78- and CD19- /CD78+).
- Animals are considered chimeric when at least 5% of WBCs in all of these lineages are donor derived.
- the systemic immune status of humanized mice is assessed pre-HSC transplantation and at week 4 and week 8 post-transplant by carboxyfluorescein diacetate succinimidyl ester (CFSE) mixed lymphocyte reaction (MLR) proliferation assays.
- CFSE carboxyfluorescein diacetate succinimidyl ester
- MLR mixed lymphocyte reaction
- MLR results are compared before PMBC-HSC transplantation and after study end point.
- [00374] Determine the efficacy of pMHC-CAR T-effector cells on specific depletion of recipient alloreactive T cells and establishment of Mixed Chimerism.
- Described herein is a flexible CAL and/or CAR design that can target any alloantigen specific T cells. Due to the diversity of alloantigen-specific T cells in the context of transplantation, the CAL and/or CAR system that has the flexibility to locate and attack different alloreactive T cells simultaneously. CAL and/or CAR T-eff. cells with (donor) pMHC can target alloreactive T-cells.
- CAL and/or CAR T cell therapy on targeting a double antigen-MHC system (OTi and OTii)
- pMHC multimer attached to adaptor molecule e.g., the CAL
- CAR T effector cell is used in the single and double specific antigen-MHC systems.
- the antigen-specific CAL and/or CAR system can be tested in vitro and in the OTi & ii transgenic mouse model.
- This model is genetically engineered such that all T cells express receptors that are specific for recognizing chicken ovalbumin peptides (257-264 (OTi) and 329-337 (OTii) in the context of H2Kb (OTi) and I-A b (OTii)).
- the T cells of these mice models are engineered to express TCRs that specifically recognize pMHC class I / II + Ovalbumin peptides. More importantly, this highly controlled model can be leveraged establish correlation between T cell activities and CAL and/or CAR T cell experimental parameters, such coculture conditions, pMHC affinity, pMHC multimer concentration.
- MHC multimers are commercially available and can be readily conjugated with FITC labeling to be recognized by the anti FITC CAR T cells.
- FITC labeling to be recognized by the anti FITC CAR T cells.
- Minor Histocompatibility Antigens are short immunogenic peptides originating from digested intra / extracellular proteins presented by Major Histocompatibility Complex (human leukocyte antigen). Disparities in minor histocompatibility antigens between individuals who are even MHC matched can induce an immune response after transplantation.
- sequences of the peptides from the donor-recipient disparity peptide library are generated in practical scale by commercially available services 39 and loaded on peptide-exchangeable MHC multimers (e.g., a CAL) (QuickSwitch MBL international or Biolegend FlexTetramer) to ultimately be bound to CAR T cells and used for targeting alloreactive T cells in the aforementioned in vitro and in vivo models (Fig.8, 9).
- donor and recipient MHCs associated peptides are isolated independently by immunoaffinity purification using the anti-HLA monoclonal antibodies 36 .
- Eluted peptides are identified by different LC-MS/MS systems in DIA (Data Independent A) mode.
- Mass Spectrometry output files are converted, searched, and statistically validated using software tools (NETMHC, SysteMHC).
- the identified peptides are then clustered (by GibbsCluster v.1) and annotated by length and predicted MHC binding affinity (NetMHC v.4).
- the final list of high- confidence donor / recipient MHC-associated peptides is compared between the recipient and the donor and the disparate sequences used for building high-quality donor-recipient disparity peptide library, which is employed as a source for generating peptides to be combined with MBL QuickSwitch (or Bio-legend FlexT) MHC tetramer system.
- PBMC / HSCT mouse model Using the same double humanized PBMC / HSCT mouse model the efficacy of this donor/recipient disparity peptide library + MHCis evaluated for specific targeting of alloreactive memory T cells and skin allograft survival.
- Hu-PBMC treated with exchanged peptide MHC (e.g., CAL) + CAR T Hu-PBMC treated with exchanged peptide MHC (e.g., CAL) + CAR T.
- HSCT with fetal HSC is performed and 6 weeks later skin from the same HSC donor is grafted on the back of the mice. Presence of mixed chimerism and graft survival is evaluated using the criteria that was mentioned earlier herein.
- the immunopeptidome library is novel and has not previously been adapted for the context of transplantation.
- Host alloreactive memory T cells influence tolerance to kidney allografts in nonhuman primates. Sci. Transl. Med.3, (2011). 18. Lo, D. J. et al. Selective targeting of human alloresponsive CD8+ effector memory T cells based on CD2 expression. Am. J. Transplant.11, 22–33 (2011). 19. Ng, Z. Y., Read, C., Kurtz, J. M. & Cetrulo, C. L. Memory T Cells in Vascularized Composite Allotransplantation. Vasc. Compos. Allotransplantation 2, 75–79 (2015). 20. Tonsho, M. et al.
- Human memory T cells Generation, compartmentalization and homeostasis. Nature Reviews Immunology vol.1424–35 (2014). 30. Thome, J. J. C. et al. Spatial map of human t cell compartmentalization and maintenance over decades of life. Cell 159, 814–828 (2014). 31. Antibodies directed against cd127. 32. Yong, K. S. M., Her, Z. & Chen, Q. Humanized Mice as Unique Tools for Human-Specific Studies. Archivum Immunologiae et Therapiae Experimentalis (2016) doi:10.1007/s00005-018-0506- x. 33. Xia, J. et al. Modeling Human Leukemia Immunotherapy in Humanized Mice.
- FITC-conjugated tetramer mediated activation was verified (Fig.12) and followed in a time course (Fig.13).
- Anti-FITC Jurkat vs OT-I experiments were conducted using 100k cells anti- FITC CAR Jurkat, 100k cells splenocytes from OT-I mouse, and 5 ug/mL to 78 ng/mL FITC- conjugated tetramer (positive, H2Kb-SIINFEKL (SEQ ID NO: 2750); negative, I-Ab-AAHAEINEA (SEQ ID NO: 2751)).
- Total Jurkat cell counts at 24 hours (Fig.14) and tetramer staining levels (Fig. 15) were also determined.
- tetramer and CD69 were also determined (Fig.16).
- This experiment demonstrated that Jurkat cells get activated in a dose dependent fashion up to 1 ug/ml of tetramer and OTi binding and staining is dose dependent. No cytotoxicity effects were seen on the killer cells and no prominent activation was seen on the target cells.
- Primary T cells – OTi [00402] Experimental design is shown in Fig.17. Human CD8 pMHC-CAR T killing is highly specific and MHC-CAR T cell does not kill bystander CD4 T cells (Fig.18). No cytotoxicity effect was seen on human CD8 pMHC- CAR T (Fig.19).
- the 1E6 clone mediates ⁇ -cell-specific killing via recognition of a highly distinctive HLA A*0201-presented signal peptide epitope (PPI15-24) that exhibits glucose-dependent presentation on the surface of human ⁇ -cells (9).
- PPI15-24 highly distinctive HLA A*0201-presented signal peptide epitope
- the inventors generated and tested a split pMHC- CAR that targets 1E6 T-cells through A*0201+peptides. Effective killing of 1E6 T-cells was observed (Fig.29)
- Example 11 [00405] CALs comprising ovalbumin MiHA (against OTi or OTii) were constructed (Fig.26).
- pMHC tetramer + FITC adaptor binds to the target cells (OTi) in a dose dependent fashion, while OTii specific tetramer does not (Fig.27). Binding of Jurkat+ pMHC to OTi cells does not change Jurkat live count (Fig.28B and 30). However, human CD8 CAR T were cytotoxic with high specificity (Fig.31 and 32). Cytotoxicity was not seen with ctrl tetramer. No cytotoxicity was seen on killer CAR T cells or bystander CD4 T Cells after co-culturing with pMHC and splenocytes.
Abstract
Description
Claims
Priority Applications (6)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CA3151961A CA3151961A1 (en) | 2019-10-18 | 2020-10-16 | Cal-t constructs and uses thereof |
CN202080087925.8A CN115175935A (en) | 2019-10-18 | 2020-10-16 | CAL-T constructs and uses thereof |
EP20876353.2A EP4093768A4 (en) | 2019-10-18 | 2020-10-16 | Cal-t constructs and uses thereof |
JP2022522834A JP2023500799A (en) | 2019-10-18 | 2020-10-16 | CAL-T constructs and their uses |
IL291976A IL291976A (en) | 2019-10-18 | 2020-10-16 | Cal-t constructs and uses thereof |
AU2020366434A AU2020366434A1 (en) | 2019-10-18 | 2020-10-16 | CAL-T constructs and uses thereof |
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US201962916924P | 2019-10-18 | 2019-10-18 | |
US62/916,924 | 2019-10-18 |
Publications (1)
Publication Number | Publication Date |
---|---|
WO2021076887A1 true WO2021076887A1 (en) | 2021-04-22 |
Family
ID=75492397
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/US2020/055980 WO2021076887A1 (en) | 2019-10-18 | 2020-10-16 | Cal-t constructs and uses thereof |
Country Status (8)
Country | Link |
---|---|
US (2) | US11723950B2 (en) |
EP (1) | EP4093768A4 (en) |
JP (1) | JP2023500799A (en) |
CN (1) | CN115175935A (en) |
AU (1) | AU2020366434A1 (en) |
CA (1) | CA3151961A1 (en) |
IL (1) | IL291976A (en) |
WO (1) | WO2021076887A1 (en) |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2021222227A1 (en) * | 2020-04-27 | 2021-11-04 | Memorial Sloan-Kettering Cancer Center | Chimeric antigen receptors targeting cd127 and use thereof |
Families Citing this family (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US11539518B2 (en) * | 2017-05-17 | 2022-12-27 | Apple Inc. | Time-based encryption key derivation |
US11475787B2 (en) * | 2017-10-20 | 2022-10-18 | Utah Valley University | Nanotechnology fabrication in a virtual reality environment |
US11763503B2 (en) * | 2019-02-25 | 2023-09-19 | Life Impact Solutions | Media alteration based on variable geolocation metadata |
GB202113858D0 (en) * | 2021-09-28 | 2021-11-10 | Nextera As | Peptides |
CN115960253A (en) * | 2022-08-30 | 2023-04-14 | 暨南大学 | Tumor T cell epitope peptide, pMHC, preparation and application thereof |
Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2018204717A1 (en) * | 2017-05-03 | 2018-11-08 | Harpoon Therapeutics, Inc. | Compositions and methods for adoptive cell therapies |
US20180346541A1 (en) * | 2015-11-23 | 2018-12-06 | Trustees Of Boston University | Methods and compositions relating to chimeric antigen receptors |
WO2019099440A1 (en) * | 2017-11-14 | 2019-05-23 | Arcellx, Inc. | Multifunctional immune cell therapies |
Family Cites Families (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US9233125B2 (en) * | 2010-12-14 | 2016-01-12 | University Of Maryland, Baltimore | Universal anti-tag chimeric antigen receptor-expressing T cells and methods of treating cancer |
BR112014029417B1 (en) | 2012-05-25 | 2023-03-07 | Cellectis | EX VIVO METHOD FOR THE PREPARATION OF T CELLS FOR IMMUNOTHERAPY |
CA2883502C (en) | 2012-09-04 | 2021-05-04 | Cellectis | Multi-chain chimeric antigen receptor and uses thereof |
EP3151854B1 (en) | 2014-06-06 | 2020-02-26 | Memorial Sloan Kettering Cancer Center | Mesothelin-targeted chimeric antigen receptors and uses thereof |
SG11201706774WA (en) | 2015-02-27 | 2017-09-28 | Icell Gene Therapeutics Llc | Chimeric antigen receptors (cars) targeting hematologic malignancies, compositions and methods of use thereof |
-
2020
- 2020-10-16 IL IL291976A patent/IL291976A/en unknown
- 2020-10-16 EP EP20876353.2A patent/EP4093768A4/en active Pending
- 2020-10-16 CA CA3151961A patent/CA3151961A1/en active Pending
- 2020-10-16 AU AU2020366434A patent/AU2020366434A1/en active Pending
- 2020-10-16 JP JP2022522834A patent/JP2023500799A/en active Pending
- 2020-10-16 WO PCT/US2020/055980 patent/WO2021076887A1/en active Application Filing
- 2020-10-16 CN CN202080087925.8A patent/CN115175935A/en active Pending
- 2020-10-16 US US17/072,490 patent/US11723950B2/en active Active
-
2023
- 2023-06-16 US US18/336,158 patent/US20230321183A1/en active Pending
Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20180346541A1 (en) * | 2015-11-23 | 2018-12-06 | Trustees Of Boston University | Methods and compositions relating to chimeric antigen receptors |
WO2018204717A1 (en) * | 2017-05-03 | 2018-11-08 | Harpoon Therapeutics, Inc. | Compositions and methods for adoptive cell therapies |
WO2019099440A1 (en) * | 2017-11-14 | 2019-05-23 | Arcellx, Inc. | Multifunctional immune cell therapies |
Non-Patent Citations (1)
Title |
---|
See also references of EP4093768A4 * |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2021222227A1 (en) * | 2020-04-27 | 2021-11-04 | Memorial Sloan-Kettering Cancer Center | Chimeric antigen receptors targeting cd127 and use thereof |
Also Published As
Publication number | Publication date |
---|---|
CN115175935A (en) | 2022-10-11 |
EP4093768A1 (en) | 2022-11-30 |
IL291976A (en) | 2022-06-01 |
US20210113653A1 (en) | 2021-04-22 |
US11723950B2 (en) | 2023-08-15 |
US20230321183A1 (en) | 2023-10-12 |
CA3151961A1 (en) | 2021-04-22 |
AU2020366434A1 (en) | 2022-03-17 |
EP4093768A4 (en) | 2023-11-15 |
JP2023500799A (en) | 2023-01-11 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
US20230321183A1 (en) | Cal-t constructs and uses thereof | |
US11596654B2 (en) | Human leukocyte antigen restricted gamma delta T cell receptors and methods of use thereof | |
JP7281774B2 (en) | ANTI-HLA-A2 ANTIBODY AND METHOD OF USE THEREOF | |
US10751373B2 (en) | PD-L1 expressing hematopoietic stem cells and uses | |
AU2016386923B2 (en) | Therapeutic use of inhibitors of T cell activation or stimulation | |
KR20170037626A (en) | Treatment of cancer using humanized anti-bcma chimeric antigen receptor | |
JP5980508B2 (en) | Treatment methods for autoimmune disorders | |
Chen et al. | Mechanistic basis of immunotherapies for type 1 diabetes mellitus | |
JP2020532304A (en) | T cell receptors that bind to hybrid leukemia (MLL) -specific phosphopeptides and how to use them | |
US20240139242A1 (en) | Cal-t constructs and uses thereof | |
WO2020018708A1 (en) | Compositions and methods for treatment of t cell malignancies | |
US20240052006A1 (en) | Anti-inflammatory cytokines and methods of use | |
WO2018236986A1 (en) | Engineered t-cell receptors and methods of their use | |
US20220389092A1 (en) | Methods and compositions involving chimeric binding polypeptides | |
RU2782276C2 (en) | Anti-hla-a2 antibodies and their application methods |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
121 | Ep: the epo has been informed by wipo that ep was designated in this application |
Ref document number: 20876353 Country of ref document: EP Kind code of ref document: A1 |
|
ENP | Entry into the national phase |
Ref document number: 2020366434 Country of ref document: AU Date of ref document: 20201016 Kind code of ref document: A |
|
ENP | Entry into the national phase |
Ref document number: 3151961 Country of ref document: CA |
|
ENP | Entry into the national phase |
Ref document number: 2022522834 Country of ref document: JP Kind code of ref document: A |
|
WWE | Wipo information: entry into national phase |
Ref document number: 17769495 Country of ref document: US |
|
NENP | Non-entry into the national phase |
Ref country code: DE |
|
ENP | Entry into the national phase |
Ref document number: 2020876353 Country of ref document: EP Effective date: 20220518 |