CN111991551A - Traf3半胱氨酸突变体在制备抗病毒药物中的应用 - Google Patents
Traf3半胱氨酸突变体在制备抗病毒药物中的应用 Download PDFInfo
- Publication number
- CN111991551A CN111991551A CN202010759780.3A CN202010759780A CN111991551A CN 111991551 A CN111991551 A CN 111991551A CN 202010759780 A CN202010759780 A CN 202010759780A CN 111991551 A CN111991551 A CN 111991551A
- Authority
- CN
- China
- Prior art keywords
- traf3
- cysteine
- mutant
- antiviral
- cysteine mutant
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 108090000922 TNF receptor-associated factor 3 Proteins 0.000 title claims abstract description 103
- 102000004399 TNF receptor-associated factor 3 Human genes 0.000 title claims abstract description 103
- 235000018417 cysteine Nutrition 0.000 title claims abstract description 67
- XUJNEKJLAYXESH-UHFFFAOYSA-N cysteine Natural products SCC(N)C(O)=O XUJNEKJLAYXESH-UHFFFAOYSA-N 0.000 title claims abstract description 62
- 239000003443 antiviral agent Substances 0.000 title claims abstract description 11
- 230000000840 anti-viral effect Effects 0.000 claims abstract description 17
- 230000001404 mediated effect Effects 0.000 claims abstract description 11
- 230000019491 signal transduction Effects 0.000 claims abstract description 9
- 238000002360 preparation method Methods 0.000 claims abstract description 8
- 239000004475 Arginine Substances 0.000 claims abstract description 3
- ODKSFYDXXFIFQN-UHFFFAOYSA-N arginine Natural products OC(=O)C(N)CCCNC(N)=N ODKSFYDXXFIFQN-UHFFFAOYSA-N 0.000 claims abstract description 3
- 108090000623 proteins and genes Proteins 0.000 claims description 22
- 239000003814 drug Substances 0.000 claims description 14
- 102000002227 Interferon Type I Human genes 0.000 claims description 10
- 108010014726 Interferon Type I Proteins 0.000 claims description 10
- 102000004127 Cytokines Human genes 0.000 claims description 7
- 108090000695 Cytokines Proteins 0.000 claims description 7
- 230000015788 innate immune response Effects 0.000 claims description 7
- 238000004519 manufacturing process Methods 0.000 claims description 7
- 239000000203 mixture Substances 0.000 claims description 7
- 230000000770 proinflammatory effect Effects 0.000 claims description 6
- 102000008300 Mutant Proteins Human genes 0.000 claims description 5
- 108010021466 Mutant Proteins Proteins 0.000 claims description 5
- 150000001945 cysteines Chemical class 0.000 claims description 5
- 230000001737 promoting effect Effects 0.000 claims description 5
- 125000000151 cysteine group Chemical group N[C@@H](CS)C(=O)* 0.000 claims description 4
- 239000000463 material Substances 0.000 claims description 2
- 239000004480 active ingredient Substances 0.000 claims 1
- 239000008194 pharmaceutical composition Substances 0.000 claims 1
- 238000002474 experimental method Methods 0.000 abstract description 13
- 230000035772 mutation Effects 0.000 abstract description 13
- 241000700605 Viruses Species 0.000 abstract description 10
- 238000001890 transfection Methods 0.000 abstract description 8
- 238000001514 detection method Methods 0.000 abstract description 7
- 230000002757 inflammatory effect Effects 0.000 abstract description 6
- 230000029812 viral genome replication Effects 0.000 abstract description 6
- 208000035473 Communicable disease Diseases 0.000 abstract description 4
- 102000014150 Interferons Human genes 0.000 abstract description 4
- 108010050904 Interferons Proteins 0.000 abstract description 4
- 229940079322 interferon Drugs 0.000 abstract description 4
- 238000011161 development Methods 0.000 abstract description 2
- 210000004027 cell Anatomy 0.000 description 25
- 239000013612 plasmid Substances 0.000 description 19
- 108020004414 DNA Proteins 0.000 description 17
- 241000711975 Vesicular stomatitis virus Species 0.000 description 17
- 102000004169 proteins and genes Human genes 0.000 description 15
- 235000018102 proteins Nutrition 0.000 description 11
- 230000034512 ubiquitination Effects 0.000 description 9
- 238000010798 ubiquitination Methods 0.000 description 9
- 238000011144 upstream manufacturing Methods 0.000 description 9
- 208000015181 infectious disease Diseases 0.000 description 8
- 108020004999 messenger RNA Proteins 0.000 description 8
- 238000000034 method Methods 0.000 description 8
- 239000000047 product Substances 0.000 description 7
- HCHKCACWOHOZIP-UHFFFAOYSA-N Zinc Chemical compound [Zn] HCHKCACWOHOZIP-UHFFFAOYSA-N 0.000 description 6
- 229910052725 zinc Inorganic materials 0.000 description 6
- 101000665442 Homo sapiens Serine/threonine-protein kinase TBK1 Proteins 0.000 description 5
- 239000005089 Luciferase Substances 0.000 description 5
- 108091005685 RIG-I-like receptors Proteins 0.000 description 5
- 102100038192 Serine/threonine-protein kinase TBK1 Human genes 0.000 description 5
- 102000002689 Toll-like receptor Human genes 0.000 description 5
- 108020000411 Toll-like receptor Proteins 0.000 description 5
- 102000000160 Tumor Necrosis Factor Receptor-Associated Peptides and Proteins Human genes 0.000 description 5
- 108010080432 Tumor Necrosis Factor Receptor-Associated Peptides and Proteins Proteins 0.000 description 5
- 238000001262 western blot Methods 0.000 description 5
- PAYRUJLWNCNPSJ-UHFFFAOYSA-N Aniline Chemical compound NC1=CC=CC=C1 PAYRUJLWNCNPSJ-UHFFFAOYSA-N 0.000 description 4
- 102000008949 Histocompatibility Antigens Class I Human genes 0.000 description 4
- 108010088652 Histocompatibility Antigens Class I Proteins 0.000 description 4
- 101001011382 Homo sapiens Interferon regulatory factor 3 Proteins 0.000 description 4
- 102100029843 Interferon regulatory factor 3 Human genes 0.000 description 4
- 108090001005 Interleukin-6 Proteins 0.000 description 4
- 108060001084 Luciferase Proteins 0.000 description 4
- 108700008625 Reporter Genes Proteins 0.000 description 4
- 230000033228 biological regulation Effects 0.000 description 4
- 238000009835 boiling Methods 0.000 description 4
- HVYWMOMLDIMFJA-DPAQBDIFSA-N cholesterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CCCC(C)C)[C@@]1(C)CC2 HVYWMOMLDIMFJA-DPAQBDIFSA-N 0.000 description 4
- 238000000605 extraction Methods 0.000 description 4
- 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 4
- 238000003753 real-time PCR Methods 0.000 description 4
- 230000001105 regulatory effect Effects 0.000 description 4
- 239000013598 vector Substances 0.000 description 4
- 239000011701 zinc Substances 0.000 description 4
- 229920000936 Agarose Polymers 0.000 description 3
- 102000003945 NF-kappa B Human genes 0.000 description 3
- 108010057466 NF-kappa B Proteins 0.000 description 3
- 102100036673 Sterol O-acyltransferase 2 Human genes 0.000 description 3
- 230000004913 activation Effects 0.000 description 3
- 238000000749 co-immunoprecipitation Methods 0.000 description 3
- 229940079593 drug Drugs 0.000 description 3
- 230000006698 induction Effects 0.000 description 3
- 239000007788 liquid Substances 0.000 description 3
- 239000006166 lysate Substances 0.000 description 3
- 230000004048 modification Effects 0.000 description 3
- 238000012986 modification Methods 0.000 description 3
- 230000010076 replication Effects 0.000 description 3
- 238000012163 sequencing technique Methods 0.000 description 3
- 230000003612 virological effect Effects 0.000 description 3
- MZOFCQQQCNRIBI-VMXHOPILSA-N (3s)-4-[[(2s)-1-[[(2s)-1-[[(1s)-1-carboxy-2-hydroxyethyl]amino]-4-methyl-1-oxopentan-2-yl]amino]-5-(diaminomethylideneamino)-1-oxopentan-2-yl]amino]-3-[[2-[[(2s)-2,6-diaminohexanoyl]amino]acetyl]amino]-4-oxobutanoic acid Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](CC(O)=O)NC(=O)CNC(=O)[C@@H](N)CCCCN MZOFCQQQCNRIBI-VMXHOPILSA-N 0.000 description 2
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 2
- 101000837584 Homo sapiens Acetyl-CoA acetyltransferase, cytosolic Proteins 0.000 description 2
- 101000642613 Homo sapiens Sterol O-acyltransferase 2 Proteins 0.000 description 2
- 102100026720 Interferon beta Human genes 0.000 description 2
- 108090000467 Interferon-beta Proteins 0.000 description 2
- 101710120037 Toxin CcdB Proteins 0.000 description 2
- 108060008682 Tumor Necrosis Factor Proteins 0.000 description 2
- 102000000852 Tumor Necrosis Factor-alpha Human genes 0.000 description 2
- 150000001413 amino acids Chemical group 0.000 description 2
- 239000003242 anti bacterial agent Substances 0.000 description 2
- 229940088710 antibiotic agent Drugs 0.000 description 2
- 235000009697 arginine Nutrition 0.000 description 2
- 239000011324 bead Substances 0.000 description 2
- 230000005754 cellular signaling Effects 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 235000012000 cholesterol Nutrition 0.000 description 2
- 239000002299 complementary DNA Substances 0.000 description 2
- 238000010276 construction Methods 0.000 description 2
- 230000002950 deficient Effects 0.000 description 2
- 238000001962 electrophoresis Methods 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 239000013604 expression vector Substances 0.000 description 2
- 230000006870 function Effects 0.000 description 2
- 239000001963 growth medium Substances 0.000 description 2
- 230000001965 increasing effect Effects 0.000 description 2
- 230000002401 inhibitory effect Effects 0.000 description 2
- 239000012160 loading buffer Substances 0.000 description 2
- 108010089193 pattern recognition receptors Proteins 0.000 description 2
- 102000007863 pattern recognition receptors Human genes 0.000 description 2
- 238000013326 plasmid cotransfection Methods 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 239000000523 sample Substances 0.000 description 2
- 230000011664 signaling Effects 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- 239000012130 whole-cell lysate Substances 0.000 description 2
- GOZMBJCYMQQACI-UHFFFAOYSA-N 6,7-dimethyl-3-[[methyl-[2-[methyl-[[1-[3-(trifluoromethyl)phenyl]indol-3-yl]methyl]amino]ethyl]amino]methyl]chromen-4-one;dihydrochloride Chemical compound Cl.Cl.C=1OC2=CC(C)=C(C)C=C2C(=O)C=1CN(C)CCN(C)CC(C1=CC=CC=C11)=CN1C1=CC=CC(C(F)(F)F)=C1 GOZMBJCYMQQACI-UHFFFAOYSA-N 0.000 description 1
- 102000007469 Actins Human genes 0.000 description 1
- 108010085238 Actins Proteins 0.000 description 1
- 101150013553 CD40 gene Proteins 0.000 description 1
- 102000000844 Cell Surface Receptors Human genes 0.000 description 1
- 108010001857 Cell Surface Receptors Proteins 0.000 description 1
- 241000282693 Cercopithecidae Species 0.000 description 1
- 102000053602 DNA Human genes 0.000 description 1
- 229920001353 Dextrin Polymers 0.000 description 1
- 239000004375 Dextrin Substances 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 102100038546 Fibronectin type III and SPRY domain-containing protein 1 Human genes 0.000 description 1
- 102000011787 Histone Methyltransferases Human genes 0.000 description 1
- 108010036115 Histone Methyltransferases Proteins 0.000 description 1
- 101001030521 Homo sapiens Fibronectin type III and SPRY domain-containing protein 1 Proteins 0.000 description 1
- 101000616738 Homo sapiens NAD-dependent protein deacetylase sirtuin-6 Proteins 0.000 description 1
- 101000850762 Homo sapiens TNF receptor-associated factor 3 Proteins 0.000 description 1
- 101000831496 Homo sapiens Toll-like receptor 3 Proteins 0.000 description 1
- 101000669447 Homo sapiens Toll-like receptor 4 Proteins 0.000 description 1
- 241001502974 Human gammaherpesvirus 8 Species 0.000 description 1
- 102000004889 Interleukin-6 Human genes 0.000 description 1
- 102000003960 Ligases Human genes 0.000 description 1
- 108090000364 Ligases Proteins 0.000 description 1
- 102100021840 NAD-dependent protein deacetylase sirtuin-6 Human genes 0.000 description 1
- 108700020796 Oncogene Proteins 0.000 description 1
- 241000283973 Oryctolagus cuniculus Species 0.000 description 1
- 238000012408 PCR amplification Methods 0.000 description 1
- 239000012807 PCR reagent Substances 0.000 description 1
- 208000037273 Pathologic Processes Diseases 0.000 description 1
- 108091000080 Phosphotransferase Proteins 0.000 description 1
- 229940124158 Protease/peptidase inhibitor Drugs 0.000 description 1
- 238000010803 Rapid Extraction Total RNA Kit Methods 0.000 description 1
- 229920002472 Starch Polymers 0.000 description 1
- 229930006000 Sucrose Natural products 0.000 description 1
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 1
- 102100033082 TNF receptor-associated factor 3 Human genes 0.000 description 1
- 102000003714 TNF receptor-associated factor 6 Human genes 0.000 description 1
- 108090000009 TNF receptor-associated factor 6 Proteins 0.000 description 1
- 102100024324 Toll-like receptor 3 Human genes 0.000 description 1
- 102100039360 Toll-like receptor 4 Human genes 0.000 description 1
- 102100040245 Tumor necrosis factor receptor superfamily member 5 Human genes 0.000 description 1
- 102000044159 Ubiquitin Human genes 0.000 description 1
- 108090000848 Ubiquitin Proteins 0.000 description 1
- 102000006275 Ubiquitin-Protein Ligases Human genes 0.000 description 1
- 108010083111 Ubiquitin-Protein Ligases Proteins 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 150000007513 acids Chemical class 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 230000003213 activating effect Effects 0.000 description 1
- 239000013543 active substance Substances 0.000 description 1
- 239000002671 adjuvant Substances 0.000 description 1
- 235000001014 amino acid Nutrition 0.000 description 1
- 230000030741 antigen processing and presentation Effects 0.000 description 1
- 230000006907 apoptotic process Effects 0.000 description 1
- 150000001484 arginines Chemical class 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 230000022131 cell cycle Effects 0.000 description 1
- 230000030833 cell death Effects 0.000 description 1
- 239000013592 cell lysate Substances 0.000 description 1
- 230000004663 cell proliferation Effects 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- 230000007960 cellular response to stress Effects 0.000 description 1
- 230000006364 cellular survival Effects 0.000 description 1
- 239000013522 chelant Substances 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 239000005482 chemotactic factor Substances 0.000 description 1
- 150000001840 cholesterol esters Chemical class 0.000 description 1
- 238000012761 co-transfection Methods 0.000 description 1
- 230000009918 complex formation Effects 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 238000005520 cutting process Methods 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 235000019425 dextrin Nutrition 0.000 description 1
- 235000014113 dietary fatty acids Nutrition 0.000 description 1
- 230000004069 differentiation Effects 0.000 description 1
- 231100000673 dose–response relationship Toxicity 0.000 description 1
- 230000002222 downregulating effect Effects 0.000 description 1
- 230000009977 dual effect Effects 0.000 description 1
- 238000010864 dual luciferase reporter gene assay Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 210000003527 eukaryotic cell Anatomy 0.000 description 1
- 239000013613 expression plasmid Substances 0.000 description 1
- 229930195729 fatty acid Natural products 0.000 description 1
- 239000000194 fatty acid Substances 0.000 description 1
- 150000004665 fatty acids Chemical class 0.000 description 1
- 238000007710 freezing Methods 0.000 description 1
- 230000008014 freezing Effects 0.000 description 1
- 239000012737 fresh medium Substances 0.000 description 1
- 239000003292 glue Substances 0.000 description 1
- HNDVDQJCIGZPNO-UHFFFAOYSA-N histidine Natural products OC(=O)C(N)CC1=CN=CN1 HNDVDQJCIGZPNO-UHFFFAOYSA-N 0.000 description 1
- 125000000487 histidyl group Chemical group [H]N([H])C(C(=O)O*)C([H])([H])C1=C([H])N([H])C([H])=N1 0.000 description 1
- 210000002865 immune cell Anatomy 0.000 description 1
- 230000028993 immune response Effects 0.000 description 1
- 238000003119 immunoblot Methods 0.000 description 1
- 210000005007 innate immune system Anatomy 0.000 description 1
- 230000003834 intracellular effect Effects 0.000 description 1
- 230000009545 invasion Effects 0.000 description 1
- 239000003446 ligand Substances 0.000 description 1
- 230000004322 lipid homeostasis Effects 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 238000010369 molecular cloning Methods 0.000 description 1
- 230000009871 nonspecific binding Effects 0.000 description 1
- 108020004707 nucleic acids Proteins 0.000 description 1
- 102000039446 nucleic acids Human genes 0.000 description 1
- 150000007523 nucleic acids Chemical class 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 244000052769 pathogen Species 0.000 description 1
- 230000009054 pathological process Effects 0.000 description 1
- 230000037361 pathway Effects 0.000 description 1
- 239000000137 peptide hydrolase inhibitor Substances 0.000 description 1
- 230000026731 phosphorylation Effects 0.000 description 1
- 238000006366 phosphorylation reaction Methods 0.000 description 1
- 102000020233 phosphotransferase Human genes 0.000 description 1
- 230000035790 physiological processes and functions Effects 0.000 description 1
- 230000016833 positive regulation of signal transduction Effects 0.000 description 1
- 230000004481 post-translational protein modification Effects 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 230000009711 regulatory function Effects 0.000 description 1
- 238000003030 reporter gene method Methods 0.000 description 1
- 238000010839 reverse transcription Methods 0.000 description 1
- 238000003757 reverse transcription PCR Methods 0.000 description 1
- 102200043451 rs121965069 Human genes 0.000 description 1
- 102220021092 rs28897672 Human genes 0.000 description 1
- 239000012723 sample buffer Substances 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 239000008107 starch Substances 0.000 description 1
- 235000019698 starch Nutrition 0.000 description 1
- 239000007858 starting material Substances 0.000 description 1
- 108010016092 sterol O-acyltransferase 2 Proteins 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
- 230000002195 synergetic effect Effects 0.000 description 1
- 230000008685 targeting Effects 0.000 description 1
- 230000002103 transcriptional effect Effects 0.000 description 1
- 239000012096 transfection reagent Substances 0.000 description 1
- 230000009385 viral infection Effects 0.000 description 1
- 210000000605 viral structure Anatomy 0.000 description 1
- 238000001086 yeast two-hybrid system Methods 0.000 description 1
Images
Classifications
-
- 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
- A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- A61K38/177—Receptors; Cell surface antigens; Cell surface determinants
- A61K38/1793—Receptors; Cell surface antigens; Cell surface determinants for cytokines; for lymphokines; for interferons
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/12—Antivirals
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Medicinal Chemistry (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- General Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Pharmacology & Pharmacy (AREA)
- Epidemiology (AREA)
- Cell Biology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Engineering & Computer Science (AREA)
- Gastroenterology & Hepatology (AREA)
- Zoology (AREA)
- Immunology (AREA)
- Virology (AREA)
- Communicable Diseases (AREA)
- Oncology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Organic Chemistry (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
本发明涉及医学生物检测技术领域，提供了TRAF3半胱氨酸突变体在制备抗病毒药物中的应用，所述应用为TRAF3第56位及第124位半胱氨酸中的至少一个突变为精氨酸。通过细胞转染实验结果可知，TRAF3半胱氨酸突变体分子可有效促进TRAF3介导的信号转导、促进I型干扰素和炎症因子的表达，以此抑制病毒复制，从而发挥抗病毒作用。因此，本发明为TRAF3半胱氨酸突变体分子的抗病毒应用提供了新的依据，揭示了TRAF3半胱氨酸突变在病毒感染性疾病发展判断和抗病毒药物制备中的新用途，为病毒感染性疾病的治疗提供了一个潜在的新靶点，具有一定的临床应用前景。
Description
技术领域
本发明涉及生物医药领域，涉及TRAF3半胱氨酸突变体在制备抗病毒药物中的应用。
背景技术
固有免疫系统是机体抵抗病毒等外来病原体入侵的重要部分，在机体识别和清除病毒 的过程中发挥着关键作用。其对病毒成分的识别主要通过模式识别受体(Patternrecognition receptors，PRRs)，包括Toll样受体(TLRs)、RIG-I样受体(RLRs)和病毒双链DNA 识别分子等，介导免疫细胞中的信号转导分子活化，产生I型干扰素、炎症因子和趋化因 子等免疫活性介质，阻断病毒复制，保护机体免受病毒的侵害。
TRAF3是肿瘤坏死因子受体相关因子(Tumor necrosis factor receptor-associated factors, TRAFs)家族中的一员。TRAF家族具有保守的模块化结构域，包括氨基末端的RING(really interesting gene)结构域、羧基末端的TRAF结构域和中间的不同数量的锌指结构，其中 TRAF结构域使得TRAFs能与细胞表面受体或其它信号分子相互作用，在调节细胞应激 反应、生存和死亡中发挥重要作用(Bradley J R,Pober J S.Tumornecrosis factor receptor-associated factors(TRAFs).Oncogene.2001,20(44):6482-6491)。
TRAF3最早是在应用酵母双杂交系统中发现的一个可结合CD40的蛋白(Hu H M,O'Rourke K,Boguski M S,et al.A novel RING finger protein interacts with thecytoplasmic domain ofCD40.J Biol Chem.1994,269(48):30069-30072)，后续研究发现TRAF3可促进 TLR3和TLR4活化诱导的I型干扰素的产生，连接着上游的接头分子和下游相关激酶 (Hacker H,Redecke V,Blagoev B,et al.Specificity in Toll-like receptorsignalling through distinct effector functions of TRAF3 and TRAF6[J].Nature.2006,439(7073):204-207； Oganesyan G,Saha S K,Guo B,et al.Criticalrole ofTRAF3 in the Toll-like receptor-dependent and-independent antiviralresponse.Nature.2006,439(7073):208-211)；此外，TRAF3也能负 向调控TLR介导的促炎性细胞因子的表达，在TLR介导的固有免疫反应中起重要调节作 用(Yang X D,Sun SC.Targeting signaling factors for degradation,an emerging mechanism for TRAFfunctions.Immunol Rev.2015,266(1):56-71)。
TRAF3包含六个锌指结构，其中两个(RZ1和RZ2)位于RING结构域，其余四个(Z1、Z2、Z3和Z4)位于RING结构域和TRAF结构域之间。TRAF3第53位、56位、73位和 76位的半胱氨酸共同鳌合一个锌原子形成锌指结构RZ1；与此同时，第117位、124位、 141位的半胱氨酸和第136位的组氨酸共同鳌合一个锌原子形成另一个锌指结构Z1。
泛素化修饰是真核细胞生物中普遍存在的一种蛋白质翻译后修饰，广泛参与细胞的各 种生理病理过程，如细胞增殖、凋亡、细胞周期、抗原提呈、免疫应答等。TRAF3不同的泛素化方式能选择地激活I型干扰素和促炎细胞因子的表达，但目前的研究都集中于TRAF3赖氨酸残基的泛素化，关于非赖氨酸残基作为泛素化位点的研究较少，而关于半 胱氨酸残基泛素化的研究则更少。
半胱氨酸上的泛素化修饰最早是在MHC-I分子中发现的。MHC-I分子的胞内结构域缺乏赖氨酸残基，MHC-I分子被由卡波西肉瘤相关的疱疹病毒编码的E3泛素连接酶MIR1 催化半胱氨酸残基泛素化，并下调MHC-I分子的表达(Cadwell K,Coscoy L.Ubiquitinationon nonlysine residues by a viral E3 ubiquitin ligase.Science.2005,309(5731):127-130)。酰基 辅酶A：胆固醇酰基转移酶2(ACAT2)是细胞内一种将胆固醇和脂肪酸转化为胆固醇酯 的酶，当脂质水平较低时，其277位的半胱氨酸(Cys277)上发生K48位连接的多聚泛素 化而被降解，从而调节脂质稳态(Wang Y J,Bian Y,Luo J,et al.Cholesteroland fatty acids regulate cysteine ubiquitylation of ACAT2 through competitiveoxidation.Nat Cell Biol. 2017,19(7):808-819)。Suv39h1是一种H3K9me3特异性组蛋白甲基转移酶，当激活NF-κB 信号通路时，SirT6能与Suv39h1结合并诱导Suv39h1发生半胱氨酸泛素化，Suv39h1得以 从IκBα中被释放，从而抑制NF-κB信号通路，起到负向调控功能(Santos-Barriopedro I, Bosch-Presegue L,Marazuela-Duque A,et al.SIRT6-dependent cysteine monoubiquitination in the PRE-SET domain of Suv39h1regulates the NF-kappaB pathway.Nat Commun. 2018,9(1):101)。但目前尚未见TRAF3半胱氨酸泛素化或突变调控抗病毒固有免疫反应的 有关报道。
发明内容
本发明是为解决上述问题进行的，目的在于对TRAF3半胱氨酸突变体在抗病毒调控 中的作用进行探索，提供了TRAF3半胱氨酸突变在制备抗病毒药物中的应用。
本发明发现了TRAF3第56位或第124位的半胱氨酸突变后能更有效地招募TBK1 形成TRAF3-TBK1复合物，且显著增加NF-κB和IRF3荧光素酶报告基因的表达，进而 促进固有免疫中RLR信号介导的I型干扰素和促炎细胞因子的产生，从而抑制病毒的复 制，保护机体，由此进行本发明，并进一步提供了TRAF3半胱氨酸突变体在用于抗病毒 治疗的新用途。
本发明的第一方面，提供了TRAF3半胱氨酸突变体在制备抗病毒药物中的应用。
优选的，TRAF3第56位及第124位的至少一个位点的半胱氨酸突变成精氨酸。
本发明中所述的TRAF3半胱氨酸突变体包括但不限于：TRAF3半胱氨酸突变体分子、TRAF3半胱氨酸突变体分子作为活性物质的组合物、含有TRAF3半胱氨酸突变体分 子的载体(如质粒)等。
所述的TRAF3半胱氨酸突变体分子，包括TRAF3半胱氨酸突变体基因、TRAF3半 胱氨酸突变体蛋白。所述的TRAF3半胱氨酸突变体基因在对象体内经转录翻译为TRAF3 半胱氨酸突变体蛋白产物。
含有TRAF3半胱氨酸突变体分子的载体的构建方法为：由于TRAF3分子的序列在本领域中是已知的，本领域普通技术人员可基于常规手段制备或通过市售获得扩增 TRAF3cDNA的引物或TRAF3质粒。以市售或构建的TRAF3质粒为模板，通过获得扩 增TRAF3半胱氨酸突变cDNA的引物，经PCR扩增获得TRAF3半胱氨酸突变线性化载 体，在连接酶的作用下，将线性化的载体相连接，筛选连接成功的质粒并进行测序鉴定。 鉴定正确的质粒即可用于后续的应用。
本发明经细胞实验后结果显示：利用质粒点突变方法将TRAF3(human)第56位、 第124位和第61位的半胱氨酸(Cysteine，C)突变为精氨酸(Arginine，R)(C61R为对 照质粒)，再利用293T细胞共转染系统和免疫共沉淀实验，发现TRAF3 C56和C124残 基的突变不仅能促进TBK1/TRAF3复合物的形成，还能促进NF-κB和IRF3荧光素酶报 告基因的表达(图1)。
在293T细胞中共转染野生型TRAF3及其突变体质粒后，用滤泡性口炎病毒(VSV)感染细胞，与野生型TRAF3相比，TRAF3半胱氨酸突变体更能促进VSV感染诱导的I 型干扰素IFN-β和促炎细胞因子IL-6和TNF-α的mRNA表达，细胞中VSV的复制水平更 低(图2)。
为进一步验证TRAF3半胱氨酸残基对I型干扰素的调控作用，我们又将野生型TRAF3、TRAF3-C56R和TRAF3-C124R的表达质粒转染于TRAF3缺陷的293T细胞中 (TRAF3-/-)，经Western Blot实验检测内源性TRAF3的蛋白表达情况，经VSV感染诱导 后检测I型干扰素的表达。结果发现，与野生型TRAF3相比，TRAF3-C56R和 TRAF3-C124R都可促进VSV感染诱导的IFN-β的mRNA表达(图2)。
因此，本发明中抗病毒药物为为促进TRAF3介导的信号转导的药物，进而为促进固有免疫中RLR信号介导的I型干扰素和促炎细胞因子的产生的药物。
本发明所述的TRAF3半胱氨酸突变体来自：人、大鼠、小鼠、犬、马、牛、兔或猴 等。
本发明的第二方面提供了一种抗病毒药物组合物，由活性组分以及药学上可接受的 辅料组成。该活性组分包括TRAF3半胱氨酸突变体分子、承载该突变体分子的载体或包含该突变体分子的组合物；辅料包括淀粉、蔗糖、糊精等。其中，该TRAF3半胱氨酸突 变体分子包括TRAF3半胱氨酸突变体基因或TRAF3半胱氨酸突变体蛋白。
本发明的有益保障及效果如下：
通过细胞转染实验结果可知，TRAF3半胱氨酸突变体分子可有效促进TRAF3介导的信号转导、促进I型干扰素和炎症因子的表达，以此抑制病毒复制，从而发挥抗病毒作用。因此，本发明为TRAF3半胱氨酸突变体分子的抗病毒应用提供了新的依据，揭示了TRAF3 半胱氨酸突变在病毒感染性疾病发展判断和抗病毒药物制备中的新用途，为病毒感染性疾病的治疗提供了一个潜在的新靶点，具有一定的临床应用前景。
附图说明
图1：TRAF3半胱氨酸C56和C124突变体促进TRAF3相关信号通路的活化，其中： 图1a为TRAF3(human)的氨基酸序列及结构域图示；图1b为在HEK293T细胞中通过 免疫共沉淀和Western Blotting方法检测TRAF3/TBK1复合物形成；图1c、d为在HEK293 细胞中通过双荧光素酶报告基因方法检测NF-κB和IRF3的活化；结果显示平均值±标准 差(n＝3)；*，P<0.05；**，P<0.01；***，P<0.001。
图2：TRAF3半胱氨酸突变体促进I型干扰素和炎性细胞因子的mRNA表达并抑制VSV病毒复制，其中：图2a、b为在HEK293细胞中通过质粒共转染方法，经VSV感染 诱导后实时荧光定量PCR检测IFN-β、IL-6、TNF-α(a)和VSV(b)mRNA表达水平； 图2c、d为在TRAF3缺陷的293T细胞中(TRAF3-/-)通过质粒共转染，经Western Blot 实验检测内源性TRAF3的蛋白表达情况(c)，经VSV感染诱导后实时荧光定量PCR检 测IFN-βmRNA表达水平(d)。结果显示平均值±标准差(n＝3)；*，P<0.05；**，P<0.01； ***，P<0.001。
具体实施方式
现结合实施例和附图，对本发明作详细描述，但本发明的实施不仅限于此。
本发明所用试剂和原料均市售可得或可按文献方法制备。下列实施例中未注明具体条 件的实验方法，通常按照常规条件如Sambrook等人《分子克隆：实验室指南》(NewYork： Cold Spring Harbor Laboratory Press，1989)中所述的条件，或按照常规条件，或按照制造 厂商所建议的条件。除非另外说明，否则百分比和份数按体积计算。
实施例1：TRAF3-C56和C124突变体能促进TRAF3介导的信号转导
突变质粒构建与抽提：野生型TRAF3表达载体购自Origene公司，以此为模板使用Takara公司的MutanBEST kit试剂盒(货号：R401)构建TRAF3分子不同氨基酸位点突 变的表达载体。突变引物序列如下：
TRAF3-C56R：
上游：CGCCACCTGGTGCTGTGCAGCCCGAAG(SEQ ID NO.1)
下游：CTTCTCACACTTGTACTTGTCCTCCACGGT(SEQ ID NO.2)
TRAF3-C124R：
上游：CGTGCAGAGCAGTTAATGCTGGGACATCTG(SEQ ID NO.3)
下游：ACCTCTGCTTTCATTCCGACAATAGATCTG(SEQ ID NO.4)
TRAF3-C61R：
上游：CGCAGCCCGAAGCAGACCGAGTGT(SEQ ID NO.5)
下游：CAGCACCAGGTGGCACTTCTCACACTTGTA(SEQ ID NO.6)
进行PCR反应后，获得所需突变体的线性化载体。将PCR产物进行琼脂糖电泳，切下目的条带做胶回收，经磷酸化处理后进行连接产物转化。在LB平板上挑取单个克隆菌落，放入2ml含氨苄抗生素的液体LB培养基中，37℃摇床摇菌后送上海生工生物工程 有限公司测序。将测序鉴定正确的500μl菌液加入50ml含氨苄抗生素的液体LB培养基 中，37℃摇床200转/分摇约14小时，以备中量提取。使用Invitrogen PureLinkTM HipurePlasmidMidiprep Kit试剂盒(货号：K210005)进行质粒抽提，按照产品说明书操作进行。利用紫外分光光度计测定抽提质粒的浓度，保存于负20℃冰箱备用。
HEK293T细胞质粒转染：HEK293T细胞购自上海中乔新舟生物技术有限公司(细 胞最初购于美国ATCC公司)。使用的转染试剂为jetPEI(PolyPlus，101-40N)。转染前一 天将HEK293T细胞铺6cm平皿，细胞密度为1×106/孔。转染前更换新鲜培养基，按产品 说明书进行实验操作。转染24或48小时后可用于检测mRNA或蛋白表达水平。
免疫共沉淀实验：用NP-40裂解液和蛋白酶抑制剂(按1：100的比例)配成细胞裂解液，提取细胞总蛋白后，用BCA法测定蛋白浓度。每个样品取相同量的蛋白，用NP-40 裂解液配平一致体积，加入上样缓冲液沸水煮6分钟，作为全细胞裂解液(WCL)冻于 负20℃备用。剩下的样品(以1mg总蛋白量为例)加入1μg对照IgG抗体和10μl protein-G 琼脂糖珠，4℃轻摇2小时进行预沉淀，以去除非特异结合。3000g/分4℃离心5分钟， 取上清至新EP管中，加入1μg目的蛋白抗体或抗标签蛋白抗体(同时用对应种属IgG抗 体作为对照)，再加入10μl protein-G琼脂糖珠，4℃轻摇过夜。3000g/分4℃离心5分钟， 弃上清，加入1ml NP-40裂解液，4℃轻摇十分钟，重复3次。加入40μl 2×蛋白上样缓 冲液后沸水煮6分钟，冻于负20℃备用。
Western Blotting检测：提取细胞总蛋白后，使用BCA法测定蛋白浓度。取相同量的 总蛋白，配平体积一致。加入6×蛋白上样缓冲液，煮沸10分钟，通过SDS PAGE电泳， 通过免疫印迹实验使用相应抗体检测各蛋白分子的表达情况。
实验中所使用抗体为：Myc(71D10)购自Cell Signaling Technology公司。normalrabbit IgG(sc-2027)购自Santa Cruz Biotechnology公司。Flag Tag(F1804)购自Sigma公司。 Anti-Gapdh(6004)购自Proteintech公司。
双荧光素酶报告基因检测：HEK293细胞购自上海中乔新舟生物技术有限公司(细胞 最初购于美国ATCC公司)。将HEK293细胞接种于96孔培养板，细胞密度为3×104/孔， 24小时后转染质粒。质粒转染24小时后，使用双荧光素酶报告基因检测试剂盒(Promega) 进行检测，步骤按照产品说明书进行。
如图1b，TRAF3-C56和C124的突变质粒比野生型TRAF3能更有效地招募TBK1形 成TRAF3/TBK1复合物。与野生型TRAF3相比，突变型TRAF3(C56R和C124R)都能 以剂量依赖的方式显著增加NF-κB和IRF3荧光素酶报告基因的表达(图1c、d)。以上 结果说明C56和C124的突变能促进TRAF3介导的信号转导，增强下游分子的转录表达。
实施例2：TRAF3半胱氨酸突变促进I型干扰素和炎性细胞因子的产生并抑制VSV病毒复制
HEK293细胞质粒转染实验方法及步骤同前叙述。
实时荧光定量PCR检测I型干扰素和炎症因子的表达：使用RNAfast200总RNA急 速抽提试剂盒(220010，上海飞捷生物技术有限公司)提取总RNA，按照产品说明书步 骤进行。
反转录PCR使用Takara反转录试剂盒(大连宝生物工程有限公司)，程序如下：42℃，59min；72℃，15min；4℃，5min。
实时荧光定量PCR：使用Promega定量PCR试剂。
所用引物均由Sangon Biotech Co.(上海生工生物工程有限公司)合成。具体序列如 下：
β-actin：
上游：AGTGTGACGTTGACATCCGT(SEQ ID NO.7)
下游：GCAGCTCAGTAACAGTCCGC(SEQ ID NO.8)
IFN-β：
上游：ATGAGTGGTGGTTGCAGGC(SEQ ID NO.9)
下游：TGACCTTTCAAATGCAGTAGATTCA(SEQ ID NO.10)
IL-6：
上游：TAGTCCTTCCTACCCCAATTTCC(SEQ ID NO：11)
下游：TTGGTCCTTAGCCACTCCTTC(SEQ ID NO：12)
TNF-α：
上游：AAGCCTGTAGCCCACGTCGTA(SEQ ID NO.13)
下游：GGCACCACTAGTTGGTTGTCTTTG(SEQ ID NO.14)
VSV：
上游：ACGGCGTACTTCCAGATGG(SEQ ID NO.15)
下游：CTCGGTTCAAGATCCAGGT(SEQ ID NO.16)
Western Blotting检测所使用抗体TRAF3(4729)购自Cell SignalingTechnology公司。
结果如图2所示，TRAF3半胱氨酸突变能促进VSV感染诱导的IFN-β、IL-6和TNF-αmRNA的表达(图2a)，进而抑制VSV病毒的复制水平(图2b)。在野生型TRAF3、 TRAF3-C56和C124的突变质粒与内源性TRAF3表达水平相近的情况下，与野生型 TRAF3相比，TRAF3-C56和C124的突变质粒更能增加VSV感染诱导的IFN-βmRNA的 表达。以上结果说明，TRAF3-C56和C124的突变在VSV病毒诱导的固有免疫应答中发 挥了重要调控作用。
由上述实验可知，病毒VSV的核酸RNA可被机体的TLRs和RLRs等所识别，进而 促使生物机体产生固有免疫应答，据此可推知，在机体受到其他病毒感染时，病毒核酸组 分被机体识别后，通过提高机体的TRAF3半胱氨酸突变体表达量能够有效抵抗病毒感染， 抑制病毒复制，从而保护机体。
以上已对本发明创造的较佳实施例进行了具体说明，但本发明创造并不限于所述实施 例，熟悉本领域的技术人员在不违背本发明创造精神的前提下还可作出种种的等同的变型 或替换，这些等同的变型或替换均包含在本申请权利要求所限定的范围内。
序列表
<110> 中国人民解放军海军军医大学
<120> TRAF3半胱氨酸突变体在制备抗病毒药物中的应用
<130> 说明书
<160> 16
<170> SIPOSequenceListing 1.0
<210> 1
<211> 27
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 1
cgccacctgg tgctgtgcag cccgaag 27
<210> 2
<211> 30
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 2
cttctcacac ttgtacttgt cctccacggt 30
<210> 3
<211> 30
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 3
cgtgcagagc agttaatgct gggacatctg 30
<210> 4
<211> 30
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 4
acctctgctt tcattccgac aatagatctg 30
<210> 5
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 5
cgcagcccga agcagaccga gtgt 24
<210> 6
<211> 30
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 6
cagcaccagg tggcacttct cacacttgta 30
<210> 7
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 7
agtgtgacgt tgacatccgt 20
<210> 8
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 8
gcagctcagt aacagtccgc 20
<210> 9
<211> 19
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 9
atgagtggtg gttgcaggc 19
<210> 10
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 10
tgacctttca aatgcagtag attca 25
<210> 11
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 11
tagtccttcc taccccaatt tcc 23
<210> 12
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 12
ttggtcctta gccactcctt c 21
<210> 13
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 13
aagcctgtag cccacgtcgt a 21
<210> 14
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 14
ggcaccacta gttggttgtc tttg 24
<210> 15
<211> 19
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 15
acggcgtact tccagatgg 19
<210> 16
<211> 19
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 16
ctcggttcaa gatccaggt 19
Claims (8)
1.TRAF3半胱氨酸突变体在制备抗病毒药物中的应用，其特征在于，TRAF3第56位及第124位半胱氨酸中的至少一个发生突变。
2.根据权利要求1所述的TRAF3半胱氨酸突变体在制备抗病毒药物中的应用，其特征在于，TRAF3第56位或第124位的半胱氨酸突变成精氨酸。
3.根据权利要求1所述的TRAF3半胱氨酸突变体在制备抗病毒药物中的应用，其特征在于，所述药物以TRAF3半胱氨酸突变体分子、承载该突变体分子的载体或包含该突变体分子的组合物为活性组分。
4.根据权利要求2所述的TRAF3半胱氨酸突变体在制备抗病毒药物中的应用，其特征在于，所述TRAF3半胱氨酸突变体分子包括TRAF3半胱氨酸突变体基因或TRAF3半胱氨酸突变体蛋白。
5.根据权利要求1所述的TRAF3半胱氨酸突变体在制备抗病毒药物中的应用，其特征在于，所述抗病毒药物为促进固有免疫中RLR信号介导的I型干扰素和促炎细胞因子的产生的药物。
6.根据权利要求1所述的TRAF3半胱氨酸突变体在制备抗病毒药物中的应用，其特征在于，所述抗病毒药物为促进TRAF3介导的信号转导的药物。
7.一种抗病毒药物组合物，其特征在于，由活性组分以及药学上可接受的辅料组成，所述活性组分为TRAF3半胱氨酸突变体分子、承载该突变体分子的载体或包含该突变体分子的组合物。
8.根据权利要求7所述的抗病毒药物组合物，其特征在于，TRAF3半胱氨酸突变体分子包括TRAF3半胱氨酸突变体基因或TRAF3半胱氨酸突变体蛋白。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010759780.3A CN111991551A (zh) | 2020-07-31 | 2020-07-31 | Traf3半胱氨酸突变体在制备抗病毒药物中的应用 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010759780.3A CN111991551A (zh) | 2020-07-31 | 2020-07-31 | Traf3半胱氨酸突变体在制备抗病毒药物中的应用 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN111991551A true CN111991551A (zh) | 2020-11-27 |
Family
ID=73462636
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202010759780.3A Pending CN111991551A (zh) | 2020-07-31 | 2020-07-31 | Traf3半胱氨酸突变体在制备抗病毒药物中的应用 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN111991551A (zh) |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2000023590A2 (en) * | 1998-10-20 | 2000-04-27 | Georgetown University Medical Center | INHIBITION OF CD40-MEDIATED NFλB ACTIVATION |
| EP1274840A2 (en) * | 2000-04-12 | 2003-01-15 | La Jolla Institute For Allergy And Immunology | Ligand for herpes simplex virus entry mediator and methods of use |
| WO2010123365A1 (en) * | 2009-04-24 | 2010-10-28 | Academisch Ziekenhuis Leiden H.O.D.N. Lumc | Increasing the immunogenicity of epithelial cells infected with human papilloma virus (hpv) |
-
2020
- 2020-07-31 CN CN202010759780.3A patent/CN111991551A/zh active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2000023590A2 (en) * | 1998-10-20 | 2000-04-27 | Georgetown University Medical Center | INHIBITION OF CD40-MEDIATED NFλB ACTIVATION |
| EP1274840A2 (en) * | 2000-04-12 | 2003-01-15 | La Jolla Institute For Allergy And Immunology | Ligand for herpes simplex virus entry mediator and methods of use |
| WO2010123365A1 (en) * | 2009-04-24 | 2010-10-28 | Academisch Ziekenhuis Leiden H.O.D.N. Lumc | Increasing the immunogenicity of epithelial cells infected with human papilloma virus (hpv) |
Non-Patent Citations (3)
| Title |
|---|
| PENG GAO ET AL.: "E3 ligase Nedd4l promotes antiviral innate immunity by catalyzing K29-linked cysteine ubiquitination of TRAF3" * |
| 刘国科: "泛素连接酶Nedd4l对巨噬细胞Toll样受体信号转导的影响" * |
| 袁铭: "E3泛素连接酶NEDD4L在抗病毒固有免疫应答中的调控及机制研究" * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Mohanty et al. | Mechanisms of oncogenesis by HTLV-1 Tax | |
| Cao et al. | The protooncogene c-Maf is an essential transcription factor for IL-10 gene expression in macrophages | |
| Bentz et al. | Epstein–Barr virus BRLF1 inhibits transcription of IRF3 and IRF7 and suppresses induction of interferon-β | |
| EP2591797A1 (en) | Regulatory factor of foxp3 and regulatory t cells and use thereof | |
| US20170189496A1 (en) | Use of phosphorylation pathway-related factor in regulating function of regulatory t cell | |
| CN105327350A (zh) | 泛素化途径相关因子在调控辅助性t细胞功能中的应用 | |
| CN110420331B (zh) | Alkbh5抑制物在治疗病毒感染性疾病中的应用 | |
| Urin et al. | CRISPR/Cas9-based knockout strategy elucidates components essential for type 1 interferon signaling in human HeLa cells | |
| Sampson et al. | The roles of E3 ubiquitin ligases in cancer progression and targeted therapy | |
| Sun et al. | An E3 ubiquitin ligase TRIM9 is involved in WSSV infection via interaction with β-TrCP | |
| Perčulija et al. | Diverse roles of DEAD/DEAH-box helicases in innate immunity and diseases | |
| Hsu et al. | The anti-proliferative effects of type I IFN involve STAT6-mediated regulation of SP1 and BCL6 | |
| Wu et al. | Grouper TRAF4, a novel, CP-interacting protein that promotes red-spotted grouper nervous necrosis virus replication | |
| Schroeder et al. | ZNF395 is an activator of a subset of IFN‐stimulated genes | |
| Matsuda | The physiological and pathophysiological role of IL-6/STAT3-mediated signal transduction and STAT3 binding partners in therapeutic applications | |
| CN111588840A (zh) | 组蛋白去泛素化酶在制备治疗系统性红斑狼疮药物的应用 | |
| Siebenlist et al. | NF-κB: A mediator of pathogen and stress responses | |
| Ahsan et al. | Multipronged regulation of autophagy and apoptosis: emerging role of TRIM proteins | |
| CN111991551A (zh) | Traf3半胱氨酸突变体在制备抗病毒药物中的应用 | |
| SEN | Transcriptional regulation of interferon-inducible genes | |
| Yang et al. | Interferon induces miR-21 through a STAT3-dependent pathway as a suppressive negative feedback on interferon-induced apoptosis | |
| Li et al. | Essential contribution of the JAK/STAT pathway to carcinogenesis, lytic infection of herpesviruses and pathogenesis of COVID‑19 | |
| Choi et al. | Novel Functions and Virus–Host Interactions Implicated in Pathogenesis and Replication of Human Herpesvirus 8 | |
| CN111139299B (zh) | Josd2蛋白在制备治疗恶性肿瘤药物中的应用 | |
| CN109420171A (zh) | Rnf2在治疗病毒感染或病毒感染相关的炎症性疾病中的用途 |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20201127 |
|
| WD01 | Invention patent application deemed withdrawn after publication |