CN115044567B - 一种肺癌的代谢免疫靶点il4i1及其应用 - Google Patents
一种肺癌的代谢免疫靶点il4i1及其应用 Download PDFInfo
- Publication number
- CN115044567B CN115044567B CN202210360734.5A CN202210360734A CN115044567B CN 115044567 B CN115044567 B CN 115044567B CN 202210360734 A CN202210360734 A CN 202210360734A CN 115044567 B CN115044567 B CN 115044567B
- Authority
- CN
- China
- Prior art keywords
- il4i1
- tumor
- lung adenocarcinoma
- seq
- expression
- 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.)
- Active
Links
- 101001121408 Homo sapiens L-amino-acid oxidase Proteins 0.000 title claims abstract description 18
- 102100026388 L-amino-acid oxidase Human genes 0.000 title claims abstract 17
- 230000002503 metabolic effect Effects 0.000 title abstract description 10
- 206010058467 Lung neoplasm malignant Diseases 0.000 title abstract description 8
- 201000005202 lung cancer Diseases 0.000 title abstract description 8
- 208000020816 lung neoplasm Diseases 0.000 title abstract description 8
- 206010028980 Neoplasm Diseases 0.000 claims abstract description 62
- 208000010507 Adenocarcinoma of Lung Diseases 0.000 claims abstract description 61
- 201000005249 lung adenocarcinoma Diseases 0.000 claims abstract description 61
- 230000014509 gene expression Effects 0.000 claims abstract description 49
- 239000003814 drug Substances 0.000 claims abstract description 20
- 230000002401 inhibitory effect Effects 0.000 claims abstract description 9
- 230000006870 function Effects 0.000 claims description 17
- 239000000523 sample Substances 0.000 claims description 13
- 210000001744 T-lymphocyte Anatomy 0.000 claims description 10
- 239000003153 chemical reaction reagent Substances 0.000 claims description 10
- 238000009169 immunotherapy Methods 0.000 claims description 10
- 239000003550 marker Substances 0.000 claims description 10
- 229940079593 drug Drugs 0.000 claims description 9
- 230000000694 effects Effects 0.000 claims description 8
- 239000003795 chemical substances by application Substances 0.000 claims description 7
- 230000001737 promoting effect Effects 0.000 claims description 7
- 230000001575 pathological effect Effects 0.000 claims description 5
- 238000012216 screening Methods 0.000 claims description 5
- 230000009471 action Effects 0.000 claims description 4
- 230000005975 antitumor immune response Effects 0.000 claims description 4
- 210000005259 peripheral blood Anatomy 0.000 claims description 4
- 239000011886 peripheral blood Substances 0.000 claims description 4
- 108010017384 Blood Proteins Proteins 0.000 claims description 3
- 102000004506 Blood Proteins Human genes 0.000 claims description 3
- 206010027476 Metastases Diseases 0.000 claims description 3
- 230000006044 T cell activation Effects 0.000 claims description 3
- 230000033540 T cell apoptotic process Effects 0.000 claims description 3
- 230000006052 T cell proliferation Effects 0.000 claims description 3
- 206010064390 Tumour invasion Diseases 0.000 claims description 3
- 230000002776 aggregation Effects 0.000 claims description 3
- 238000004220 aggregation Methods 0.000 claims description 3
- 230000009400 cancer invasion Effects 0.000 claims description 3
- 238000002648 combination therapy Methods 0.000 claims description 3
- 239000012636 effector Substances 0.000 claims description 3
- 230000002708 enhancing effect Effects 0.000 claims description 3
- 230000001744 histochemical effect Effects 0.000 claims description 3
- 230000009401 metastasis Effects 0.000 claims description 3
- 210000004985 myeloid-derived suppressor cell Anatomy 0.000 claims description 3
- 108091033319 polynucleotide Proteins 0.000 claims description 3
- 239000002157 polynucleotide Substances 0.000 claims description 3
- 102000040430 polynucleotide Human genes 0.000 claims description 3
- 239000012474 protein marker Substances 0.000 claims description 3
- 230000004614 tumor growth Effects 0.000 claims description 3
- 230000005909 tumor killing Effects 0.000 claims description 3
- 125000003275 alpha amino acid group Chemical group 0.000 claims 4
- 230000002452 interceptive effect Effects 0.000 claims 1
- 210000004027 cell Anatomy 0.000 abstract description 60
- 108090000623 proteins and genes Proteins 0.000 abstract description 30
- 102000004169 proteins and genes Human genes 0.000 abstract description 23
- 201000011510 cancer Diseases 0.000 abstract description 18
- 238000013508 migration Methods 0.000 abstract description 11
- 108020004999 messenger RNA Proteins 0.000 abstract description 10
- 210000002540 macrophage Anatomy 0.000 abstract description 9
- 230000005012 migration Effects 0.000 abstract description 9
- 230000001105 regulatory effect Effects 0.000 abstract description 8
- QIVBCDIJIAJPQS-VIFPVBQESA-N L-tryptophane Chemical compound C1=CC=C2C(C[C@H](N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-VIFPVBQESA-N 0.000 abstract description 7
- QIVBCDIJIAJPQS-UHFFFAOYSA-N Tryptophan Natural products C1=CC=C2C(CC(N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-UHFFFAOYSA-N 0.000 abstract description 7
- 238000011282 treatment Methods 0.000 abstract description 7
- 210000004443 dendritic cell Anatomy 0.000 abstract description 6
- 230000001506 immunosuppresive effect Effects 0.000 abstract description 4
- 230000015572 biosynthetic process Effects 0.000 abstract description 3
- 210000001185 bone marrow Anatomy 0.000 abstract description 3
- 230000009545 invasion Effects 0.000 abstract description 3
- 239000003596 drug target Substances 0.000 abstract description 2
- 238000010837 poor prognosis Methods 0.000 abstract description 2
- 210000001519 tissue Anatomy 0.000 description 37
- 150000001413 amino acids Chemical class 0.000 description 35
- 238000000034 method Methods 0.000 description 14
- 230000002018 overexpression Effects 0.000 description 14
- 239000013612 plasmid Substances 0.000 description 13
- 108091027967 Small hairpin RNA Proteins 0.000 description 11
- 239000004055 small Interfering RNA Substances 0.000 description 11
- 239000007787 solid Substances 0.000 description 11
- 238000012163 sequencing technique Methods 0.000 description 10
- 210000004881 tumor cell Anatomy 0.000 description 10
- 238000003197 gene knockdown Methods 0.000 description 9
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 8
- BDAGIHXWWSANSR-UHFFFAOYSA-N methanoic acid Natural products OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 description 8
- 230000019491 signal transduction Effects 0.000 description 8
- 238000011529 RT qPCR Methods 0.000 description 7
- 239000002773 nucleotide Substances 0.000 description 7
- 125000003729 nucleotide group Chemical group 0.000 description 7
- 239000012188 paraffin wax Substances 0.000 description 7
- 108090000765 processed proteins & peptides Proteins 0.000 description 7
- 230000002829 reductive effect Effects 0.000 description 7
- 230000012292 cell migration Effects 0.000 description 6
- 239000002299 complementary DNA Substances 0.000 description 6
- 238000001514 detection method Methods 0.000 description 6
- 230000002441 reversible effect Effects 0.000 description 6
- 238000010276 construction Methods 0.000 description 5
- 239000013598 vector Substances 0.000 description 5
- OSWFIVFLDKOXQC-UHFFFAOYSA-N 4-(3-methoxyphenyl)aniline Chemical compound COC1=CC=CC(C=2C=CC(N)=CC=2)=C1 OSWFIVFLDKOXQC-UHFFFAOYSA-N 0.000 description 4
- 102000004190 Enzymes Human genes 0.000 description 4
- 108090000790 Enzymes Proteins 0.000 description 4
- 241000283973 Oryctolagus cuniculus Species 0.000 description 4
- 102000007079 Peptide Fragments Human genes 0.000 description 4
- 108010033276 Peptide Fragments Proteins 0.000 description 4
- 229940088598 enzyme Drugs 0.000 description 4
- 239000003797 essential amino acid Substances 0.000 description 4
- 238000002474 experimental method Methods 0.000 description 4
- 235000019253 formic acid Nutrition 0.000 description 4
- 230000002055 immunohistochemical effect Effects 0.000 description 4
- 230000004060 metabolic process Effects 0.000 description 4
- 230000004048 modification Effects 0.000 description 4
- 238000012986 modification Methods 0.000 description 4
- 238000010827 pathological analysis Methods 0.000 description 4
- 239000006228 supernatant Substances 0.000 description 4
- 238000004885 tandem mass spectrometry Methods 0.000 description 4
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 3
- 102000004388 Interleukin-4 Human genes 0.000 description 3
- 108090000978 Interleukin-4 Proteins 0.000 description 3
- 108010008292 L-Amino Acid Oxidase Proteins 0.000 description 3
- 102000007070 L-amino-acid oxidase Human genes 0.000 description 3
- 108091030071 RNAI Proteins 0.000 description 3
- 230000004913 activation Effects 0.000 description 3
- 238000006243 chemical reaction Methods 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- VWWQXMAJTJZDQX-UYBVJOGSSA-N flavin adenine dinucleotide Chemical compound C1=NC2=C(N)N=CN=C2N1[C@@H]([C@H](O)[C@@H]1O)O[C@@H]1CO[P@](O)(=O)O[P@@](O)(=O)OC[C@@H](O)[C@@H](O)[C@@H](O)CN1C2=NC(=O)NC(=O)C2=NC2=C1C=C(C)C(C)=C2 VWWQXMAJTJZDQX-UYBVJOGSSA-N 0.000 description 3
- 235000019162 flavin adenine dinucleotide Nutrition 0.000 description 3
- 239000011714 flavin adenine dinucleotide Substances 0.000 description 3
- 229940093632 flavin-adenine dinucleotide Drugs 0.000 description 3
- 230000009368 gene silencing by RNA Effects 0.000 description 3
- 238000007490 hematoxylin and eosin (H&E) staining Methods 0.000 description 3
- 210000002865 immune cell Anatomy 0.000 description 3
- 150000002500 ions Chemical class 0.000 description 3
- 238000004811 liquid chromatography Methods 0.000 description 3
- 238000004949 mass spectrometry Methods 0.000 description 3
- 210000001616 monocyte Anatomy 0.000 description 3
- 238000012545 processing Methods 0.000 description 3
- 238000010186 staining Methods 0.000 description 3
- 238000012795 verification Methods 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- 239000004475 Arginine Substances 0.000 description 2
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 208000010839 B-cell chronic lymphocytic leukemia Diseases 0.000 description 2
- IUZGUFAJDBHQQV-YUMQZZPRSA-N Gly-Leu-Asn Chemical compound NCC(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(N)=O)C(O)=O IUZGUFAJDBHQQV-YUMQZZPRSA-N 0.000 description 2
- 101000934372 Homo sapiens Macrosialin Proteins 0.000 description 2
- YOKVEHGYYQEQOP-QWRGUYRKSA-N Leu-Leu-Gly Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)NCC(O)=O YOKVEHGYYQEQOP-QWRGUYRKSA-N 0.000 description 2
- 208000031422 Lymphocytic Chronic B-Cell Leukemia Diseases 0.000 description 2
- 102100025136 Macrosialin Human genes 0.000 description 2
- 102000011779 Nitric Oxide Synthase Type II Human genes 0.000 description 2
- 108010076864 Nitric Oxide Synthase Type II Proteins 0.000 description 2
- 238000012408 PCR amplification Methods 0.000 description 2
- DWYAUVCQDTZIJI-VZFHVOOUSA-N Thr-Ala-Ser Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@@H](CO)C(O)=O DWYAUVCQDTZIJI-VZFHVOOUSA-N 0.000 description 2
- 102000004142 Trypsin Human genes 0.000 description 2
- 108090000631 Trypsin Proteins 0.000 description 2
- 101710136122 Tryptophan 2,3-dioxygenase Proteins 0.000 description 2
- 102000057288 Tryptophan 2,3-dioxygenases Human genes 0.000 description 2
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 description 2
- 208000009956 adenocarcinoma Diseases 0.000 description 2
- 229940024606 amino acid Drugs 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- ODKSFYDXXFIFQN-UHFFFAOYSA-N arginine Natural products OC(=O)C(N)CCCNC(N)=N ODKSFYDXXFIFQN-UHFFFAOYSA-N 0.000 description 2
- 108010060035 arginylproline Proteins 0.000 description 2
- 210000003719 b-lymphocyte Anatomy 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 230000033228 biological regulation Effects 0.000 description 2
- 210000004369 blood Anatomy 0.000 description 2
- 239000008280 blood Substances 0.000 description 2
- 230000004709 cell invasion Effects 0.000 description 2
- 239000006285 cell suspension Substances 0.000 description 2
- 238000005119 centrifugation Methods 0.000 description 2
- 210000000349 chromosome Anatomy 0.000 description 2
- 208000032852 chronic lymphocytic leukemia Diseases 0.000 description 2
- 238000012258 culturing Methods 0.000 description 2
- 239000008380 degradant Substances 0.000 description 2
- 230000036425 denaturation Effects 0.000 description 2
- 238000004925 denaturation Methods 0.000 description 2
- 230000001419 dependent effect Effects 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 238000003745 diagnosis Methods 0.000 description 2
- FSXRLASFHBWESK-UHFFFAOYSA-N dipeptide phenylalanyl-tyrosine Natural products C=1C=C(O)C=CC=1CC(C(O)=O)NC(=O)C(N)CC1=CC=CC=C1 FSXRLASFHBWESK-UHFFFAOYSA-N 0.000 description 2
- 238000009826 distribution Methods 0.000 description 2
- 239000002552 dosage form Substances 0.000 description 2
- 235000020776 essential amino acid Nutrition 0.000 description 2
- 108010049041 glutamylalanine Proteins 0.000 description 2
- 108010020688 glycylhistidine Proteins 0.000 description 2
- 108010050848 glycylleucine Proteins 0.000 description 2
- 238000000227 grinding Methods 0.000 description 2
- 230000006801 homologous recombination Effects 0.000 description 2
- 238000002744 homologous recombination Methods 0.000 description 2
- 230000036737 immune function Effects 0.000 description 2
- 230000036039 immunity Effects 0.000 description 2
- 238000003119 immunoblot Methods 0.000 description 2
- 239000003112 inhibitor Substances 0.000 description 2
- 238000005468 ion implantation Methods 0.000 description 2
- QWTDNUCVQCZILF-UHFFFAOYSA-N isopentane Chemical compound CCC(C)C QWTDNUCVQCZILF-UHFFFAOYSA-N 0.000 description 2
- BTNMPGBKDVTSJY-UHFFFAOYSA-N keto-phenylpyruvic acid Chemical compound OC(=O)C(=O)CC1=CC=CC=C1 BTNMPGBKDVTSJY-UHFFFAOYSA-N 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 238000001819 mass spectrum Methods 0.000 description 2
- 201000001441 melanoma Diseases 0.000 description 2
- 230000007102 metabolic function Effects 0.000 description 2
- 238000002156 mixing Methods 0.000 description 2
- 230000007170 pathology Effects 0.000 description 2
- 238000002360 preparation method Methods 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 102000004196 processed proteins & peptides Human genes 0.000 description 2
- 108010070643 prolylglutamic acid Proteins 0.000 description 2
- 238000011865 proteolysis targeting chimera technique Methods 0.000 description 2
- 229940124823 proteolysis targeting chimeric molecule Drugs 0.000 description 2
- RXWNCPJZOCPEPQ-NVWDDTSBSA-N puromycin Chemical compound C1=CC(OC)=CC=C1C[C@H](N)C(=O)N[C@H]1[C@@H](O)[C@H](N2C3=NC=NC(=C3N=C2)N(C)C)O[C@@H]1CO RXWNCPJZOCPEPQ-NVWDDTSBSA-N 0.000 description 2
- 238000000926 separation method Methods 0.000 description 2
- 150000003384 small molecules Chemical class 0.000 description 2
- 108010026668 snake venom protein C activator Proteins 0.000 description 2
- 230000002194 synthesizing effect Effects 0.000 description 2
- 238000013519 translation Methods 0.000 description 2
- 239000012588 trypsin Substances 0.000 description 2
- 238000010200 validation analysis Methods 0.000 description 2
- IHWDSEPNZDYMNF-UHFFFAOYSA-N 1H-indol-2-amine Chemical compound C1=CC=C2NC(N)=CC2=C1 IHWDSEPNZDYMNF-UHFFFAOYSA-N 0.000 description 1
- 239000001903 2-oxo-3-phenylpropanoic acid Substances 0.000 description 1
- FWBHETKCLVMNFS-UHFFFAOYSA-N 4',6-Diamino-2-phenylindol Chemical compound C1=CC(C(=N)N)=CC=C1C1=CC2=CC=C(C(N)=N)C=C2N1 FWBHETKCLVMNFS-UHFFFAOYSA-N 0.000 description 1
- 101150026173 ARG2 gene Proteins 0.000 description 1
- GSCLWXDNIMNIJE-ZLUOBGJFSA-N Ala-Asp-Asn Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(N)=O)C(O)=O GSCLWXDNIMNIJE-ZLUOBGJFSA-N 0.000 description 1
- FUSPCLTUKXQREV-ACZMJKKPSA-N Ala-Glu-Ala Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](C)C(O)=O FUSPCLTUKXQREV-ACZMJKKPSA-N 0.000 description 1
- NWVVKQZOVSTDBQ-CIUDSAMLSA-N Ala-Glu-Arg Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O NWVVKQZOVSTDBQ-CIUDSAMLSA-N 0.000 description 1
- BTBUEVAGZCKULD-XPUUQOCRSA-N Ala-Gly-His Chemical compound C[C@H](N)C(=O)NCC(=O)N[C@H](C(O)=O)CC1=CN=CN1 BTBUEVAGZCKULD-XPUUQOCRSA-N 0.000 description 1
- RZZMZYZXNJRPOJ-BJDJZHNGSA-N Ala-Ile-Lys Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCCCN)C(=O)O)NC(=O)[C@H](C)N RZZMZYZXNJRPOJ-BJDJZHNGSA-N 0.000 description 1
- YHKANGMVQWRMAP-DCAQKATOSA-N Ala-Leu-Arg Chemical compound C[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@H](C(O)=O)CCCN=C(N)N YHKANGMVQWRMAP-DCAQKATOSA-N 0.000 description 1
- HHRAXZAYZFFRAM-CIUDSAMLSA-N Ala-Leu-Asn Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(N)=O)C(O)=O HHRAXZAYZFFRAM-CIUDSAMLSA-N 0.000 description 1
- MNZHHDPWDWQJCQ-YUMQZZPRSA-N Ala-Leu-Gly Chemical compound C[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)NCC(O)=O MNZHHDPWDWQJCQ-YUMQZZPRSA-N 0.000 description 1
- MFMDKJIPHSWSBM-GUBZILKMSA-N Ala-Lys-Glu Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCC(O)=O)C(O)=O MFMDKJIPHSWSBM-GUBZILKMSA-N 0.000 description 1
- GKAZXNDATBWNBI-DCAQKATOSA-N Ala-Met-Lys Chemical compound C[C@@H](C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CCCCN)C(=O)O)N GKAZXNDATBWNBI-DCAQKATOSA-N 0.000 description 1
- GFEDXKNBZMPEDM-KZVJFYERSA-N Ala-Met-Thr Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CCSC)C(=O)N[C@@H]([C@@H](C)O)C(O)=O GFEDXKNBZMPEDM-KZVJFYERSA-N 0.000 description 1
- YYAVDNKUWLAFCV-ACZMJKKPSA-N Ala-Ser-Gln Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCC(N)=O)C(O)=O YYAVDNKUWLAFCV-ACZMJKKPSA-N 0.000 description 1
- ZCUFMRIQCPNOHZ-NRPADANISA-N Ala-Val-Gln Chemical compound C[C@@H](C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCC(=O)N)C(=O)O)N ZCUFMRIQCPNOHZ-NRPADANISA-N 0.000 description 1
- 241000372033 Andromeda Species 0.000 description 1
- PBSOQGZLPFVXPU-YUMQZZPRSA-N Arg-Glu-Gly Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(O)=O)C(=O)NCC(O)=O PBSOQGZLPFVXPU-YUMQZZPRSA-N 0.000 description 1
- UAOSDDXCTBIPCA-QXEWZRGKSA-N Arg-Ile-Gly Chemical compound CC[C@H](C)[C@@H](C(=O)NCC(=O)O)NC(=O)[C@H](CCCN=C(N)N)N UAOSDDXCTBIPCA-QXEWZRGKSA-N 0.000 description 1
- OOIMKQRCPJBGPD-XUXIUFHCSA-N Arg-Ile-Leu Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC(C)C)C(O)=O OOIMKQRCPJBGPD-XUXIUFHCSA-N 0.000 description 1
- OFIYLHVAAJYRBC-HJWJTTGWSA-N Arg-Ile-Phe Chemical compound CC[C@H](C)[C@H](NC(=O)[C@@H](N)CCCNC(N)=N)C(=O)N[C@@H](Cc1ccccc1)C(O)=O OFIYLHVAAJYRBC-HJWJTTGWSA-N 0.000 description 1
- FNXCAFKDGBROCU-STECZYCISA-N Arg-Ile-Tyr Chemical compound NC(N)=NCCC[C@H](N)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@H](C(O)=O)CC1=CC=C(O)C=C1 FNXCAFKDGBROCU-STECZYCISA-N 0.000 description 1
- WMEVEPXNCMKNGH-IHRRRGAJSA-N Arg-Leu-His Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CC1=CN=CN1)C(=O)O)NC(=O)[C@H](CCCN=C(N)N)N WMEVEPXNCMKNGH-IHRRRGAJSA-N 0.000 description 1
- NGTYEHIRESTSRX-UWVGGRQHSA-N Arg-Lys-Gly Chemical compound NCCCC[C@@H](C(=O)NCC(O)=O)NC(=O)[C@@H](N)CCCN=C(N)N NGTYEHIRESTSRX-UWVGGRQHSA-N 0.000 description 1
- LCBSSOCDWUTQQV-SDDRHHMPSA-N Arg-Met-Pro Chemical compound CSCC[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CCCN=C(N)N)N LCBSSOCDWUTQQV-SDDRHHMPSA-N 0.000 description 1
- RYQSYXFGFOTJDJ-RHYQMDGZSA-N Arg-Thr-Leu Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(C)C)C(O)=O RYQSYXFGFOTJDJ-RHYQMDGZSA-N 0.000 description 1
- DRDWXKWUSIKKOB-PJODQICGSA-N Arg-Trp-Ala Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC1=CNC2=C1C=CC=C2)C(=O)N[C@@H](C)C(O)=O DRDWXKWUSIKKOB-PJODQICGSA-N 0.000 description 1
- FMYQECOAIFGQGU-CYDGBPFRSA-N Arg-Val-Ile Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O FMYQECOAIFGQGU-CYDGBPFRSA-N 0.000 description 1
- 102000004452 Arginase Human genes 0.000 description 1
- 108700024123 Arginases Proteins 0.000 description 1
- WLKVEEODTPQPLI-ACZMJKKPSA-N Asp-Gln-Asn Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(N)=O)C(O)=O WLKVEEODTPQPLI-ACZMJKKPSA-N 0.000 description 1
- CJUKAWUWBZCTDQ-SRVKXCTJSA-N Asp-Leu-Lys Chemical compound OC(=O)C[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCCN)C(O)=O CJUKAWUWBZCTDQ-SRVKXCTJSA-N 0.000 description 1
- JXGJJQJHXHXJQF-CIUDSAMLSA-N Asp-Met-Glu Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CCC(O)=O)C(O)=O JXGJJQJHXHXJQF-CIUDSAMLSA-N 0.000 description 1
- KESWRFKUZRUTAH-FXQIFTODSA-N Asp-Pro-Asp Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CC(O)=O)C(O)=O KESWRFKUZRUTAH-FXQIFTODSA-N 0.000 description 1
- FIRWLDUOFOULCA-XIRDDKMYSA-N Asp-Trp-Lys Chemical compound C1=CC=C2C(=C1)C(=CN2)C[C@@H](C(=O)N[C@@H](CCCCN)C(=O)O)NC(=O)[C@H](CC(=O)O)N FIRWLDUOFOULCA-XIRDDKMYSA-N 0.000 description 1
- GXIUDSXIUSTSLO-QXEWZRGKSA-N Asp-Val-Met Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CCSC)C(=O)O)NC(=O)[C@H](CC(=O)O)N GXIUDSXIUSTSLO-QXEWZRGKSA-N 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- 108010029697 CD40 Ligand Proteins 0.000 description 1
- 102100032937 CD40 ligand Human genes 0.000 description 1
- KXDHJXZQYSOELW-UHFFFAOYSA-M Carbamate Chemical compound NC([O-])=O KXDHJXZQYSOELW-UHFFFAOYSA-M 0.000 description 1
- NLCZGISONIGRQP-DCAQKATOSA-N Cys-Arg-Lys Chemical compound C(CCN)C[C@@H](C(=O)O)NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](CS)N NLCZGISONIGRQP-DCAQKATOSA-N 0.000 description 1
- PGBLJHDDKCVSTC-CIUDSAMLSA-N Cys-Met-Gln Chemical compound CSCC[C@H](NC(=O)[C@@H](N)CS)C(=O)N[C@@H](CCC(N)=O)C(O)=O PGBLJHDDKCVSTC-CIUDSAMLSA-N 0.000 description 1
- 102000012410 DNA Ligases Human genes 0.000 description 1
- 108010061982 DNA Ligases Proteins 0.000 description 1
- KVYVOGYEMPEXBT-GUBZILKMSA-N Gln-Ala-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)[C@@H](N)CCC(N)=O KVYVOGYEMPEXBT-GUBZILKMSA-N 0.000 description 1
- GMGKDVVBSVVKCT-NUMRIWBASA-N Gln-Asn-Thr Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O GMGKDVVBSVVKCT-NUMRIWBASA-N 0.000 description 1
- MAGNEQBFSBREJL-DCAQKATOSA-N Gln-Glu-Lys Chemical compound C(CCN)C[C@@H](C(=O)O)NC(=O)[C@H](CCC(=O)O)NC(=O)[C@H](CCC(=O)N)N MAGNEQBFSBREJL-DCAQKATOSA-N 0.000 description 1
- CLPQUWHBWXFJOX-BQBZGAKWSA-N Gln-Gly-Gln Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)NCC(=O)N[C@@H](CCC(N)=O)C(O)=O CLPQUWHBWXFJOX-BQBZGAKWSA-N 0.000 description 1
- FGYPOQPQTUNESW-IUCAKERBSA-N Gln-Gly-Leu Chemical compound CC(C)C[C@@H](C(=O)O)NC(=O)CNC(=O)[C@H](CCC(=O)N)N FGYPOQPQTUNESW-IUCAKERBSA-N 0.000 description 1
- NSORZJXKUQFEKL-JGVFFNPUSA-N Gln-Gly-Pro Chemical compound C1C[C@@H](N(C1)C(=O)CNC(=O)[C@H](CCC(=O)N)N)C(=O)O NSORZJXKUQFEKL-JGVFFNPUSA-N 0.000 description 1
- VAIWPXWHWAPYDF-FXQIFTODSA-N Glu-Asp-Gln Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CCC(N)=O)C(O)=O VAIWPXWHWAPYDF-FXQIFTODSA-N 0.000 description 1
- IESFZVCAVACGPH-PEFMBERDSA-N Glu-Asp-Ile Chemical compound CC[C@H](C)[C@@H](C(O)=O)NC(=O)[C@H](CC(O)=O)NC(=O)[C@@H](N)CCC(O)=O IESFZVCAVACGPH-PEFMBERDSA-N 0.000 description 1
- APHGWLWMOXGZRL-DCAQKATOSA-N Glu-Glu-His Chemical compound N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](Cc1cnc[nH]1)C(O)=O APHGWLWMOXGZRL-DCAQKATOSA-N 0.000 description 1
- UHVIQGKBMXEVGN-WDSKDSINSA-N Glu-Gly-Asn Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)NCC(=O)N[C@@H](CC(N)=O)C(O)=O UHVIQGKBMXEVGN-WDSKDSINSA-N 0.000 description 1
- OAGVHWYIBZMWLA-YFKPBYRVSA-N Glu-Gly-Gly Chemical compound OC(=O)CC[C@H](N)C(=O)NCC(=O)NCC(O)=O OAGVHWYIBZMWLA-YFKPBYRVSA-N 0.000 description 1
- ILWHFUZZCFYSKT-AVGNSLFASA-N Glu-Lys-Leu Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(O)=O ILWHFUZZCFYSKT-AVGNSLFASA-N 0.000 description 1
- HZISRJBYZAODRV-XQXXSGGOSA-N Glu-Thr-Ala Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C)C(O)=O HZISRJBYZAODRV-XQXXSGGOSA-N 0.000 description 1
- ZALGPUWUVHOGAE-GVXVVHGQSA-N Glu-Val-His Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC1=CN=CN1)C(=O)O)NC(=O)[C@H](CCC(=O)O)N ZALGPUWUVHOGAE-GVXVVHGQSA-N 0.000 description 1
- ZYRXTRTUCAVNBQ-GVXVVHGQSA-N Glu-Val-Lys Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CCCCN)C(=O)O)NC(=O)[C@H](CCC(=O)O)N ZYRXTRTUCAVNBQ-GVXVVHGQSA-N 0.000 description 1
- PHONXOACARQMPM-BQBZGAKWSA-N Gly-Ala-Met Chemical compound [H]NCC(=O)N[C@@H](C)C(=O)N[C@@H](CCSC)C(O)=O PHONXOACARQMPM-BQBZGAKWSA-N 0.000 description 1
- BYYNJRSNDARRBX-YFKPBYRVSA-N Gly-Gln-Gly Chemical compound NCC(=O)N[C@@H](CCC(N)=O)C(=O)NCC(O)=O BYYNJRSNDARRBX-YFKPBYRVSA-N 0.000 description 1
- FEUPVVCGQLNXNP-IRXDYDNUSA-N Gly-Phe-Phe Chemical compound C([C@H](NC(=O)CN)C(=O)N[C@@H](CC=1C=CC=CC=1)C(O)=O)C1=CC=CC=C1 FEUPVVCGQLNXNP-IRXDYDNUSA-N 0.000 description 1
- VDCRBJACQKOSMS-JSGCOSHPSA-N Gly-Phe-Val Chemical compound [H]NCC(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](C(C)C)C(O)=O VDCRBJACQKOSMS-JSGCOSHPSA-N 0.000 description 1
- GGLIDLCEPDHEJO-BQBZGAKWSA-N Gly-Pro-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@@H]1CCCN1C(=O)CN GGLIDLCEPDHEJO-BQBZGAKWSA-N 0.000 description 1
- MKIAPEZXQDILRR-YUMQZZPRSA-N Gly-Ser-His Chemical compound C1=C(NC=N1)C[C@@H](C(=O)O)NC(=O)[C@H](CO)NC(=O)CN MKIAPEZXQDILRR-YUMQZZPRSA-N 0.000 description 1
- JQFILXICXLDTRR-FBCQKBJTSA-N Gly-Thr-Gly Chemical compound NCC(=O)N[C@@H]([C@H](O)C)C(=O)NCC(O)=O JQFILXICXLDTRR-FBCQKBJTSA-N 0.000 description 1
- MREVELMMFOLESM-HOCLYGCPSA-N Gly-Trp-Val Chemical compound [H]NCC(=O)N[C@@H](CC1=CNC2=C1C=CC=C2)C(=O)N[C@@H](C(C)C)C(O)=O MREVELMMFOLESM-HOCLYGCPSA-N 0.000 description 1
- UIQGJYUEQDOODF-KWQFWETISA-N Gly-Tyr-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@@H](NC(=O)CN)CC1=CC=C(O)C=C1 UIQGJYUEQDOODF-KWQFWETISA-N 0.000 description 1
- 206010018691 Granuloma Diseases 0.000 description 1
- DZMVESFTHXSSPZ-XVYDVKMFSA-N His-Ala-Ser Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](C)C(=O)N[C@@H](CO)C(O)=O DZMVESFTHXSSPZ-XVYDVKMFSA-N 0.000 description 1
- UOAVQQRILDGZEN-SRVKXCTJSA-N His-Asp-Leu Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(C)C)C(O)=O UOAVQQRILDGZEN-SRVKXCTJSA-N 0.000 description 1
- JFFAPRNXXLRINI-NHCYSSNCSA-N His-Asp-Val Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](C(C)C)C(O)=O JFFAPRNXXLRINI-NHCYSSNCSA-N 0.000 description 1
- UXSATKFPUVZVDK-KKUMJFAQSA-N His-Lys-Leu Chemical compound CC(C)C[C@@H](C(=O)O)NC(=O)[C@H](CCCCN)NC(=O)[C@H](CC1=CN=CN1)N UXSATKFPUVZVDK-KKUMJFAQSA-N 0.000 description 1
- CWSZWFILCNSNEX-CIUDSAMLSA-N His-Ser-Asn Chemical compound C1=C(NC=N1)C[C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(=O)N)C(=O)O)N CWSZWFILCNSNEX-CIUDSAMLSA-N 0.000 description 1
- WKEABZIITNXXQZ-CIUDSAMLSA-N His-Ser-Cys Chemical compound C1=C(NC=N1)C[C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CS)C(=O)O)N WKEABZIITNXXQZ-CIUDSAMLSA-N 0.000 description 1
- GIRSNERMXCMDBO-GARJFASQSA-N His-Ser-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CO)NC(=O)[C@H](CC2=CN=CN2)N)C(=O)O GIRSNERMXCMDBO-GARJFASQSA-N 0.000 description 1
- BRQKGRLDDDQWQJ-MBLNEYKQSA-N His-Thr-Ala Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C)C(O)=O BRQKGRLDDDQWQJ-MBLNEYKQSA-N 0.000 description 1
- FCPSGEVYIVXPPO-QTKMDUPCSA-N His-Thr-Arg Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O FCPSGEVYIVXPPO-QTKMDUPCSA-N 0.000 description 1
- CCUSLCQWVMWTIS-IXOXFDKPSA-N His-Thr-Leu Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(C)C)C(O)=O CCUSLCQWVMWTIS-IXOXFDKPSA-N 0.000 description 1
- WSXNWASHQNSMRX-GVXVVHGQSA-N His-Val-Gln Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)O)NC(=O)[C@H](CC1=CN=CN1)N WSXNWASHQNSMRX-GVXVVHGQSA-N 0.000 description 1
- 101001037256 Homo sapiens Indoleamine 2,3-dioxygenase 1 Proteins 0.000 description 1
- 101001037261 Homo sapiens Indoleamine 2,3-dioxygenase 2 Proteins 0.000 description 1
- HDODQNPMSHDXJT-GHCJXIJMSA-N Ile-Asn-Ser Chemical compound CC[C@H](C)[C@H](N)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CO)C(O)=O HDODQNPMSHDXJT-GHCJXIJMSA-N 0.000 description 1
- PNDMHTTXXPUQJH-RWRJDSDZSA-N Ile-Glu-Thr Chemical compound N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CCC(=O)O)C(=O)N[C@@H]([C@H](O)C)C(=O)O PNDMHTTXXPUQJH-RWRJDSDZSA-N 0.000 description 1
- LPFBXFILACZHIB-LAEOZQHASA-N Ile-Gly-Glu Chemical compound CC[C@H](C)[C@@H](C(=O)NCC(=O)N[C@@H](CCC(=O)O)C(=O)O)N LPFBXFILACZHIB-LAEOZQHASA-N 0.000 description 1
- HUORUFRRJHELPD-MNXVOIDGSA-N Ile-Leu-Glu Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(=O)O)C(=O)O)N HUORUFRRJHELPD-MNXVOIDGSA-N 0.000 description 1
- WXLYNEHOGRYNFU-URLPEUOOSA-N Ile-Thr-Phe Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)O)N WXLYNEHOGRYNFU-URLPEUOOSA-N 0.000 description 1
- 102100040061 Indoleamine 2,3-dioxygenase 1 Human genes 0.000 description 1
- 102100040062 Indoleamine 2,3-dioxygenase 2 Human genes 0.000 description 1
- 206010061218 Inflammation Diseases 0.000 description 1
- IBMVEYRWAWIOTN-UHFFFAOYSA-N L-Leucyl-L-Arginyl-L-Proline Natural products CC(C)CC(N)C(=O)NC(CCCN=C(N)N)C(=O)N1CCCC1C(O)=O IBMVEYRWAWIOTN-UHFFFAOYSA-N 0.000 description 1
- ODKSFYDXXFIFQN-BYPYZUCNSA-P L-argininium(2+) Chemical compound NC(=[NH2+])NCCC[C@H]([NH3+])C(O)=O ODKSFYDXXFIFQN-BYPYZUCNSA-P 0.000 description 1
- COLNVLDHVKWLRT-QMMMGPOBSA-N L-phenylalanine Chemical compound OC(=O)[C@@H](N)CC1=CC=CC=C1 COLNVLDHVKWLRT-QMMMGPOBSA-N 0.000 description 1
- BQSLGJHIAGOZCD-CIUDSAMLSA-N Leu-Ala-Ser Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@@H](CO)C(O)=O BQSLGJHIAGOZCD-CIUDSAMLSA-N 0.000 description 1
- JUWJEAPUNARGCF-DCAQKATOSA-N Leu-Arg-Ala Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](C)C(O)=O JUWJEAPUNARGCF-DCAQKATOSA-N 0.000 description 1
- HBJZFCIVFIBNSV-DCAQKATOSA-N Leu-Arg-Asn Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](CC(N)=O)C(O)=O HBJZFCIVFIBNSV-DCAQKATOSA-N 0.000 description 1
- IBMVEYRWAWIOTN-RWMBFGLXSA-N Leu-Arg-Pro Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CCCN=C(N)N)C(=O)N1CCC[C@@H]1C(O)=O IBMVEYRWAWIOTN-RWMBFGLXSA-N 0.000 description 1
- KUEVMUXNILMJTK-JYJNAYRXSA-N Leu-Gln-Tyr Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@H](C(O)=O)CC1=CC=C(O)C=C1 KUEVMUXNILMJTK-JYJNAYRXSA-N 0.000 description 1
- LLBQJYDYOLIQAI-JYJNAYRXSA-N Leu-Glu-Tyr Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O LLBQJYDYOLIQAI-JYJNAYRXSA-N 0.000 description 1
- LXKNSJLSGPNHSK-KKUMJFAQSA-N Leu-Leu-Lys Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCCN)C(=O)O)N LXKNSJLSGPNHSK-KKUMJFAQSA-N 0.000 description 1
- XVZCXCTYGHPNEM-UHFFFAOYSA-N Leu-Leu-Pro Natural products CC(C)CC(N)C(=O)NC(CC(C)C)C(=O)N1CCCC1C(O)=O XVZCXCTYGHPNEM-UHFFFAOYSA-N 0.000 description 1
- QMKFDEUJGYNFMC-AVGNSLFASA-N Leu-Pro-Arg Chemical compound CC(C)C[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CCCN=C(N)N)C(O)=O QMKFDEUJGYNFMC-AVGNSLFASA-N 0.000 description 1
- IDGZVZJLYFTXSL-DCAQKATOSA-N Leu-Ser-Arg Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@H](C(O)=O)CCCN=C(N)N IDGZVZJLYFTXSL-DCAQKATOSA-N 0.000 description 1
- IZPVWNSAVUQBGP-CIUDSAMLSA-N Leu-Ser-Asp Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(O)=O)C(O)=O IZPVWNSAVUQBGP-CIUDSAMLSA-N 0.000 description 1
- XOWMDXHFSBCAKQ-SRVKXCTJSA-N Leu-Ser-Leu Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@H](C(O)=O)CC(C)C XOWMDXHFSBCAKQ-SRVKXCTJSA-N 0.000 description 1
- IWMJFLJQHIDZQW-KKUMJFAQSA-N Leu-Ser-Phe Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 IWMJFLJQHIDZQW-KKUMJFAQSA-N 0.000 description 1
- DAYQSYGBCUKVKT-VOAKCMCISA-N Leu-Thr-Lys Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCCCN)C(O)=O DAYQSYGBCUKVKT-VOAKCMCISA-N 0.000 description 1
- LSLUTXRANSUGFY-XIRDDKMYSA-N Leu-Trp-Asp Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC1=CNC2=C1C=CC=C2)C(=O)N[C@@H](CC(O)=O)C(O)=O LSLUTXRANSUGFY-XIRDDKMYSA-N 0.000 description 1
- QUYCUALODHJQLK-CIUDSAMLSA-N Lys-Asp-Asp Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(O)=O)C(O)=O QUYCUALODHJQLK-CIUDSAMLSA-N 0.000 description 1
- ZJSZPXISKMDJKQ-JYJNAYRXSA-N Lys-Phe-Glu Chemical compound NCCCC[C@H](N)C(=O)N[C@H](C(=O)N[C@@H](CCC(O)=O)C(O)=O)CC1=CC=CC=C1 ZJSZPXISKMDJKQ-JYJNAYRXSA-N 0.000 description 1
- SVSQSPICRKBMSZ-SRVKXCTJSA-N Lys-Pro-Gln Chemical compound [H]N[C@@H](CCCCN)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CCC(N)=O)C(O)=O SVSQSPICRKBMSZ-SRVKXCTJSA-N 0.000 description 1
- OZVXDDFYCQOPFD-XQQFMLRXSA-N Lys-Val-Pro Chemical compound CC(C)[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CCCCN)N OZVXDDFYCQOPFD-XQQFMLRXSA-N 0.000 description 1
- RIPJMCFGQHGHNP-RHYQMDGZSA-N Lys-Val-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](C(C)C)NC(=O)[C@H](CCCCN)N)O RIPJMCFGQHGHNP-RHYQMDGZSA-N 0.000 description 1
- 241000124008 Mammalia Species 0.000 description 1
- FWTBMGAKKPSTBT-GUBZILKMSA-N Met-Gln-Glu Chemical compound [H]N[C@@H](CCSC)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCC(O)=O)C(O)=O FWTBMGAKKPSTBT-GUBZILKMSA-N 0.000 description 1
- ORRNBLTZBBESPN-HJWJTTGWSA-N Met-Ile-Phe Chemical compound CSCC[C@H](N)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 ORRNBLTZBBESPN-HJWJTTGWSA-N 0.000 description 1
- 101100260702 Mus musculus Tinagl1 gene Proteins 0.000 description 1
- YBAFDPFAUTYYRW-UHFFFAOYSA-N N-L-alpha-glutamyl-L-leucine Natural products CC(C)CC(C(O)=O)NC(=O)C(N)CCC(O)=O YBAFDPFAUTYYRW-UHFFFAOYSA-N 0.000 description 1
- AUEJLPRZGVVDNU-UHFFFAOYSA-N N-L-tyrosyl-L-leucine Natural products CC(C)CC(C(O)=O)NC(=O)C(N)CC1=CC=C(O)C=C1 AUEJLPRZGVVDNU-UHFFFAOYSA-N 0.000 description 1
- KZNQNBZMBZJQJO-UHFFFAOYSA-N N-glycyl-L-proline Natural products NCC(=O)N1CCCC1C(O)=O KZNQNBZMBZJQJO-UHFFFAOYSA-N 0.000 description 1
- AJHCSUXXECOXOY-UHFFFAOYSA-N N-glycyl-L-tryptophan Natural products C1=CC=C2C(CC(NC(=O)CN)C(O)=O)=CNC2=C1 AJHCSUXXECOXOY-UHFFFAOYSA-N 0.000 description 1
- BQVUABVGYYSDCJ-UHFFFAOYSA-N Nalpha-L-Leucyl-L-tryptophan Natural products C1=CC=C2C(CC(NC(=O)C(N)CC(C)C)C(O)=O)=CNC2=C1 BQVUABVGYYSDCJ-UHFFFAOYSA-N 0.000 description 1
- 108010019160 Pancreatin Proteins 0.000 description 1
- 108091005804 Peptidases Proteins 0.000 description 1
- FPTXMUIBLMGTQH-ONGXEEELSA-N Phe-Ala-Gly Chemical compound OC(=O)CNC(=O)[C@H](C)NC(=O)[C@@H](N)CC1=CC=CC=C1 FPTXMUIBLMGTQH-ONGXEEELSA-N 0.000 description 1
- BRDYYVQTEJVRQT-HRCADAONSA-N Phe-Arg-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](CC2=CC=CC=C2)N)C(=O)O BRDYYVQTEJVRQT-HRCADAONSA-N 0.000 description 1
- PSKRILMFHNIUAO-JYJNAYRXSA-N Phe-Glu-Lys Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)N[C@@H](CCCCN)C(=O)O)N PSKRILMFHNIUAO-JYJNAYRXSA-N 0.000 description 1
- YCCUXNNKXDGMAM-KKUMJFAQSA-N Phe-Leu-Ser Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(O)=O YCCUXNNKXDGMAM-KKUMJFAQSA-N 0.000 description 1
- BSTPNLNKHKBONJ-HTUGSXCWSA-N Phe-Thr-Gln Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)O)NC(=O)[C@H](CC1=CC=CC=C1)N)O BSTPNLNKHKBONJ-HTUGSXCWSA-N 0.000 description 1
- KCIKTPHTEYBXMG-BVSLBCMMSA-N Phe-Trp-Arg Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CC1=CNC2=C1C=CC=C2)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O KCIKTPHTEYBXMG-BVSLBCMMSA-N 0.000 description 1
- IFMDQWDAJUMMJC-DCAQKATOSA-N Pro-Ala-Leu Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](C)C(=O)N[C@@H](CC(C)C)C(O)=O IFMDQWDAJUMMJC-DCAQKATOSA-N 0.000 description 1
- XQLBWXHVZVBNJM-FXQIFTODSA-N Pro-Ala-Ser Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)[C@@H]1CCCN1 XQLBWXHVZVBNJM-FXQIFTODSA-N 0.000 description 1
- QBFONMUYNSNKIX-AVGNSLFASA-N Pro-Arg-His Chemical compound C1C[C@H](NC1)C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](CC2=CN=CN2)C(=O)O QBFONMUYNSNKIX-AVGNSLFASA-N 0.000 description 1
- NMELOOXSGDRBRU-YUMQZZPRSA-N Pro-Glu-Gly Chemical compound OC(=O)CNC(=O)[C@H](CCC(=O)O)NC(=O)[C@@H]1CCCN1 NMELOOXSGDRBRU-YUMQZZPRSA-N 0.000 description 1
- SBVPYBFMIGDIDX-SRVKXCTJSA-N Pro-Pro-Pro Chemical compound OC(=O)[C@@H]1CCCN1C(=O)[C@H]1N(C(=O)[C@H]2NCCC2)CCC1 SBVPYBFMIGDIDX-SRVKXCTJSA-N 0.000 description 1
- KBUAPZAZPWNYSW-SRVKXCTJSA-N Pro-Pro-Val Chemical compound CC(C)[C@@H](C(O)=O)NC(=O)[C@@H]1CCCN1C(=O)[C@H]1NCCC1 KBUAPZAZPWNYSW-SRVKXCTJSA-N 0.000 description 1
- GOMUXSCOIWIJFP-GUBZILKMSA-N Pro-Ser-Arg Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CO)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O GOMUXSCOIWIJFP-GUBZILKMSA-N 0.000 description 1
- LZHHZYDPMZEMRX-STQMWFEESA-N Pro-Tyr-Gly Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)NCC(O)=O LZHHZYDPMZEMRX-STQMWFEESA-N 0.000 description 1
- 239000004365 Protease Substances 0.000 description 1
- 108010026552 Proteome Proteins 0.000 description 1
- 238000002123 RNA extraction Methods 0.000 description 1
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 description 1
- KYKKKSWGEPFUMR-NAKRPEOUSA-N Ser-Arg-Ile Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O KYKKKSWGEPFUMR-NAKRPEOUSA-N 0.000 description 1
- ULVMNZOKDBHKKI-ACZMJKKPSA-N Ser-Gln-Asp Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(O)=O)C(O)=O ULVMNZOKDBHKKI-ACZMJKKPSA-N 0.000 description 1
- XWCYBVBLJRWOFR-WDSKDSINSA-N Ser-Gln-Gly Chemical compound OC[C@H](N)C(=O)N[C@@H](CCC(N)=O)C(=O)NCC(O)=O XWCYBVBLJRWOFR-WDSKDSINSA-N 0.000 description 1
- HJEBZBMOTCQYDN-ACZMJKKPSA-N Ser-Glu-Asp Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(O)=O)C(O)=O HJEBZBMOTCQYDN-ACZMJKKPSA-N 0.000 description 1
- IXZHZUGGKLRHJD-DCAQKATOSA-N Ser-Leu-Val Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C(C)C)C(O)=O IXZHZUGGKLRHJD-DCAQKATOSA-N 0.000 description 1
- DINQYZRMXGWWTG-GUBZILKMSA-N Ser-Pro-Pro Chemical compound OC[C@H](N)C(=O)N1CCC[C@H]1C(=O)N1[C@H](C(O)=O)CCC1 DINQYZRMXGWWTG-GUBZILKMSA-N 0.000 description 1
- AZWNCEBQZXELEZ-FXQIFTODSA-N Ser-Pro-Ser Chemical compound OC[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CO)C(O)=O AZWNCEBQZXELEZ-FXQIFTODSA-N 0.000 description 1
- NVNPWELENFJOHH-CIUDSAMLSA-N Ser-Ser-His Chemical compound C1=C(NC=N1)C[C@@H](C(=O)O)NC(=O)[C@H](CO)NC(=O)[C@H](CO)N NVNPWELENFJOHH-CIUDSAMLSA-N 0.000 description 1
- 210000000068 Th17 cell Anatomy 0.000 description 1
- VXMHQKHDKCATDV-VEVYYDQMSA-N Thr-Asp-Arg Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O VXMHQKHDKCATDV-VEVYYDQMSA-N 0.000 description 1
- YSXYEJWDHBCTDJ-DVJZZOLTSA-N Thr-Gly-Trp Chemical compound C[C@H]([C@@H](C(=O)NCC(=O)N[C@@H](CC1=CNC2=CC=CC=C21)C(=O)O)N)O YSXYEJWDHBCTDJ-DVJZZOLTSA-N 0.000 description 1
- DXPURPNJDFCKKO-RHYQMDGZSA-N Thr-Lys-Val Chemical compound CC(C)[C@H](NC(=O)[C@H](CCCCN)NC(=O)[C@@H](N)[C@@H](C)O)C(O)=O DXPURPNJDFCKKO-RHYQMDGZSA-N 0.000 description 1
- BCYUHPXBHCUYBA-CUJWVEQBSA-N Thr-Ser-His Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@@H](Cc1cnc[nH]1)C(O)=O BCYUHPXBHCUYBA-CUJWVEQBSA-N 0.000 description 1
- ZEJBJDHSQPOVJV-UAXMHLISSA-N Thr-Trp-Thr Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC1=CNC2=C1C=CC=C2)C(=O)N[C@@H]([C@@H](C)O)C(O)=O ZEJBJDHSQPOVJV-UAXMHLISSA-N 0.000 description 1
- BEZTUFWTPVOROW-KJEVXHAQSA-N Thr-Tyr-Arg Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CC1=CC=C(C=C1)O)C(=O)N[C@@H](CCCN=C(N)N)C(=O)O)N)O BEZTUFWTPVOROW-KJEVXHAQSA-N 0.000 description 1
- 102000002689 Toll-like receptor Human genes 0.000 description 1
- 108020000411 Toll-like receptor Proteins 0.000 description 1
- GTNCSPKYWCJZAC-XIRDDKMYSA-N Trp-Asp-Leu Chemical compound CC(C)C[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)O)NC(=O)[C@H](CC1=CNC2=CC=CC=C21)N GTNCSPKYWCJZAC-XIRDDKMYSA-N 0.000 description 1
- GWQUSADRQCTMHN-NWLDYVSISA-N Trp-Gln-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](CCC(=O)N)NC(=O)[C@H](CC1=CNC2=CC=CC=C21)N)O GWQUSADRQCTMHN-NWLDYVSISA-N 0.000 description 1
- STKZKWFOKOCSLW-UMPQAUOISA-N Trp-Thr-Val Chemical compound C1=CC=C2C(C[C@H](N)C(=O)N[C@H](C(=O)N[C@@H](C(C)C)C(O)=O)[C@@H](C)O)=CNC2=C1 STKZKWFOKOCSLW-UMPQAUOISA-N 0.000 description 1
- JFDGVHXRCKEBAU-KKUMJFAQSA-N Tyr-Asp-Lys Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)N[C@@H](CCCCN)C(=O)O)N)O JFDGVHXRCKEBAU-KKUMJFAQSA-N 0.000 description 1
- DXUVJJRTVACXSO-KKUMJFAQSA-N Tyr-Gln-Met Chemical compound CSCC[C@@H](C(=O)O)NC(=O)[C@H](CCC(=O)N)NC(=O)[C@H](CC1=CC=C(C=C1)O)N DXUVJJRTVACXSO-KKUMJFAQSA-N 0.000 description 1
- WAPFQMXRSDEGOE-IHRRRGAJSA-N Tyr-Glu-Gln Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(N)=O)C(O)=O WAPFQMXRSDEGOE-IHRRRGAJSA-N 0.000 description 1
- QHONGSVIVOFKAC-ULQDDVLXSA-N Tyr-Pro-His Chemical compound N[C@@H](Cc1ccc(O)cc1)C(=O)N1CCC[C@H]1C(=O)N[C@@H](Cc1cnc[nH]1)C(O)=O QHONGSVIVOFKAC-ULQDDVLXSA-N 0.000 description 1
- AKKYBQGHUAWPJR-MNSWYVGCSA-N Tyr-Thr-Trp Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CC1=CNC2=CC=CC=C21)C(=O)O)NC(=O)[C@H](CC3=CC=C(C=C3)O)N)O AKKYBQGHUAWPJR-MNSWYVGCSA-N 0.000 description 1
- OBKOPLHSRDATFO-XHSDSOJGSA-N Tyr-Val-Pro Chemical compound CC(C)[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CC2=CC=C(C=C2)O)N OBKOPLHSRDATFO-XHSDSOJGSA-N 0.000 description 1
- DJIJBQYBDKGDIS-JYJNAYRXSA-N Tyr-Val-Val Chemical compound CC(C)[C@H](NC(=O)[C@@H](NC(=O)[C@@H](N)Cc1ccc(O)cc1)C(C)C)C(O)=O DJIJBQYBDKGDIS-JYJNAYRXSA-N 0.000 description 1
- 108010064997 VPY tripeptide Proteins 0.000 description 1
- AZSHAZJLOZQYAY-FXQIFTODSA-N Val-Ala-Ser Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@@H](CO)C(O)=O AZSHAZJLOZQYAY-FXQIFTODSA-N 0.000 description 1
- AGKDVLSDNSTLFA-UMNHJUIQSA-N Val-Gln-Pro Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)N1CCC[C@@H]1C(=O)O)N AGKDVLSDNSTLFA-UMNHJUIQSA-N 0.000 description 1
- PIFJAFRUVWZRKR-QMMMGPOBSA-N Val-Gly-Gly Chemical compound CC(C)[C@H]([NH3+])C(=O)NCC(=O)NCC([O-])=O PIFJAFRUVWZRKR-QMMMGPOBSA-N 0.000 description 1
- PTFPUAXGIKTVNN-ONGXEEELSA-N Val-His-Gly Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC1=CN=CN1)C(=O)NCC(=O)O)N PTFPUAXGIKTVNN-ONGXEEELSA-N 0.000 description 1
- GVJUTBOZZBTBIG-AVGNSLFASA-N Val-Lys-Arg Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCCN=C(N)N)C(=O)O)N GVJUTBOZZBTBIG-AVGNSLFASA-N 0.000 description 1
- JAKHAONCJJZVHT-DCAQKATOSA-N Val-Lys-Ser Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CO)C(=O)O)N JAKHAONCJJZVHT-DCAQKATOSA-N 0.000 description 1
- UEXPMFIAZZHEAD-HSHDSVGOSA-N Val-Thr-Trp Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CC1=CNC2=CC=CC=C21)C(=O)O)NC(=O)[C@H](C(C)C)N)O UEXPMFIAZZHEAD-HSHDSVGOSA-N 0.000 description 1
- AOILQMZPNLUXCM-AVGNSLFASA-N Val-Val-Lys Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)N[C@H](C(O)=O)CCCCN AOILQMZPNLUXCM-AVGNSLFASA-N 0.000 description 1
- 241000700605 Viruses Species 0.000 description 1
- 230000021736 acetylation Effects 0.000 description 1
- 238000006640 acetylation reaction Methods 0.000 description 1
- 230000006978 adaptation Effects 0.000 description 1
- 230000033289 adaptive immune response Effects 0.000 description 1
- 238000000246 agarose gel electrophoresis Methods 0.000 description 1
- 108010008685 alanyl-glutamyl-aspartic acid Proteins 0.000 description 1
- 108010086434 alanyl-seryl-glycine Proteins 0.000 description 1
- 108010070944 alanylhistidine Proteins 0.000 description 1
- KOSRFJWDECSPRO-UHFFFAOYSA-N alpha-L-glutamyl-L-glutamic acid Natural products OC(=O)CCC(N)C(=O)NC(CCC(O)=O)C(O)=O KOSRFJWDECSPRO-UHFFFAOYSA-N 0.000 description 1
- DEDGUGJNLNLJSR-UHFFFAOYSA-N alpha-hydroxycinnamic acid Natural products OC(=O)C(O)=CC1=CC=CC=C1 DEDGUGJNLNLJSR-UHFFFAOYSA-N 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
- 238000000137 annealing Methods 0.000 description 1
- 230000000844 anti-bacterial effect Effects 0.000 description 1
- 230000002924 anti-infective effect Effects 0.000 description 1
- 239000000427 antigen Substances 0.000 description 1
- 108091007433 antigens Proteins 0.000 description 1
- 102000036639 antigens Human genes 0.000 description 1
- 230000006907 apoptotic process Effects 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 101150088826 arg1 gene Proteins 0.000 description 1
- 108010052670 arginyl-glutamyl-glutamic acid Proteins 0.000 description 1
- 108010038850 arginyl-isoleucyl-tyrosine Proteins 0.000 description 1
- 108010093581 aspartyl-proline Proteins 0.000 description 1
- 108010047857 aspartylglycine Proteins 0.000 description 1
- 108010068265 aspartyltyrosine Proteins 0.000 description 1
- 230000004888 barrier function Effects 0.000 description 1
- 239000000090 biomarker Substances 0.000 description 1
- 210000004899 c-terminal region Anatomy 0.000 description 1
- 239000004202 carbamide Substances 0.000 description 1
- 230000001925 catabolic effect Effects 0.000 description 1
- 210000003169 central nervous system Anatomy 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 230000001684 chronic effect Effects 0.000 description 1
- 238000003776 cleavage reaction Methods 0.000 description 1
- 230000008045 co-localization Effects 0.000 description 1
- 238000012761 co-transfection Methods 0.000 description 1
- 238000013170 computed tomography imaging Methods 0.000 description 1
- 239000012141 concentrate Substances 0.000 description 1
- 238000004624 confocal microscopy Methods 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 239000013078 crystal Substances 0.000 description 1
- 239000012228 culture supernatant Substances 0.000 description 1
- 238000005520 cutting process Methods 0.000 description 1
- 230000001351 cycling effect Effects 0.000 description 1
- 210000000805 cytoplasm Anatomy 0.000 description 1
- 230000001086 cytosolic effect Effects 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 239000008367 deionised water Substances 0.000 description 1
- 229910021641 deionized water Inorganic materials 0.000 description 1
- 230000002939 deleterious effect Effects 0.000 description 1
- 238000011033 desalting Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 230000029087 digestion Effects 0.000 description 1
- AFABGHUZZDYHJO-UHFFFAOYSA-N dimethyl butane Natural products CCCC(C)C AFABGHUZZDYHJO-UHFFFAOYSA-N 0.000 description 1
- 238000010494 dissociation reaction Methods 0.000 description 1
- 230000005593 dissociations Effects 0.000 description 1
- 239000012154 double-distilled water Substances 0.000 description 1
- 238000007877 drug screening Methods 0.000 description 1
- 238000010828 elution Methods 0.000 description 1
- 210000002919 epithelial cell Anatomy 0.000 description 1
- 210000000981 epithelium Anatomy 0.000 description 1
- 230000007717 exclusion Effects 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 239000000706 filtrate Substances 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 239000012634 fragment Substances 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
- 108010006664 gamma-glutamyl-glycyl-glycine Proteins 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 108010042598 glutamyl-aspartyl-glycine Proteins 0.000 description 1
- 230000013595 glycosylation Effects 0.000 description 1
- 238000006206 glycosylation reaction Methods 0.000 description 1
- 108010000434 glycyl-alanyl-leucine Proteins 0.000 description 1
- 108010077515 glycylproline Proteins 0.000 description 1
- 108010084389 glycyltryptophan Proteins 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 108010036413 histidylglycine Proteins 0.000 description 1
- 102000056209 human IL4I1 Human genes 0.000 description 1
- 238000010166 immunofluorescence Methods 0.000 description 1
- 238000013115 immunohistochemical detection Methods 0.000 description 1
- 238000012151 immunohistochemical method Methods 0.000 description 1
- 238000011532 immunohistochemical staining Methods 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 230000002757 inflammatory effect Effects 0.000 description 1
- 230000004054 inflammatory process Effects 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 229940028885 interleukin-4 Drugs 0.000 description 1
- 210000002570 interstitial cell Anatomy 0.000 description 1
- 238000005040 ion trap Methods 0.000 description 1
- 108010044374 isoleucyl-tyrosine Proteins 0.000 description 1
- 108010034529 leucyl-lysine Proteins 0.000 description 1
- 108010090333 leucyl-lysyl-proline Proteins 0.000 description 1
- 210000000265 leukocyte Anatomy 0.000 description 1
- 239000003446 ligand Substances 0.000 description 1
- 230000000670 limiting effect Effects 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 230000004807 localization Effects 0.000 description 1
- 210000004072 lung Anatomy 0.000 description 1
- 210000004698 lymphocyte Anatomy 0.000 description 1
- 210000003563 lymphoid tissue Anatomy 0.000 description 1
- 239000006166 lysate Substances 0.000 description 1
- 108010064235 lysylglycine Proteins 0.000 description 1
- 108010054155 lysyllysine Proteins 0.000 description 1
- 108010038320 lysylphenylalanine Proteins 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 239000011159 matrix material Substances 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 239000003147 molecular marker Substances 0.000 description 1
- 210000004877 mucosa Anatomy 0.000 description 1
- 210000000822 natural killer cell Anatomy 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 238000010899 nucleation Methods 0.000 description 1
- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- 210000004940 nucleus Anatomy 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 229940055695 pancreatin Drugs 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 239000008188 pellet Substances 0.000 description 1
- COLNVLDHVKWLRT-UHFFFAOYSA-N phenylalanine Natural products OC(=O)C(N)CC1=CC=CC=C1 COLNVLDHVKWLRT-UHFFFAOYSA-N 0.000 description 1
- 229960005190 phenylalanine Drugs 0.000 description 1
- 150000002993 phenylalanine derivatives Chemical class 0.000 description 1
- 108010078226 phenylalanine oxidase Proteins 0.000 description 1
- 108010051242 phenylalanylserine Proteins 0.000 description 1
- 239000013600 plasmid vector Substances 0.000 description 1
- 239000011148 porous material Substances 0.000 description 1
- 230000002980 postoperative effect Effects 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 238000012257 pre-denaturation Methods 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 238000004393 prognosis Methods 0.000 description 1
- 230000035755 proliferation Effects 0.000 description 1
- 108010093296 prolyl-prolyl-alanine Proteins 0.000 description 1
- 108010031719 prolyl-serine Proteins 0.000 description 1
- 108010004914 prolylarginine Proteins 0.000 description 1
- 229950010131 puromycin Drugs 0.000 description 1
- 239000013062 quality control Sample Substances 0.000 description 1
- 238000003908 quality control method Methods 0.000 description 1
- 238000003753 real-time PCR Methods 0.000 description 1
- 108020003175 receptors Proteins 0.000 description 1
- 102000005962 receptors Human genes 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 238000006722 reduction reaction Methods 0.000 description 1
- 238000005932 reductive alkylation reaction Methods 0.000 description 1
- 239000011347 resin Substances 0.000 description 1
- 229920005989 resin Polymers 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 108091008146 restriction endonucleases Proteins 0.000 description 1
- 238000010839 reverse transcription Methods 0.000 description 1
- 201000000306 sarcoidosis Diseases 0.000 description 1
- 230000007017 scission Effects 0.000 description 1
- 108010048818 seryl-histidine Proteins 0.000 description 1
- 108010026333 seryl-proline Proteins 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
- 210000000952 spleen Anatomy 0.000 description 1
- 230000000638 stimulation Effects 0.000 description 1
- 230000004960 subcellular localization Effects 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 239000013589 supplement Substances 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 201000000596 systemic lupus erythematosus Diseases 0.000 description 1
- 231100000732 tissue residue Toxicity 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 238000013518 transcription Methods 0.000 description 1
- 230000035897 transcription Effects 0.000 description 1
- 238000001890 transfection Methods 0.000 description 1
- 230000007704 transition Effects 0.000 description 1
- 238000011269 treatment regimen Methods 0.000 description 1
- 108010038745 tryptophylglycine Proteins 0.000 description 1
- 201000008827 tuberculosis Diseases 0.000 description 1
- 108010078580 tyrosylleucine Proteins 0.000 description 1
- 238000000108 ultra-filtration Methods 0.000 description 1
- 238000001291 vacuum drying Methods 0.000 description 1
- 230000003612 virological effect Effects 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/0004—Oxidoreductases (1.)
- C12N9/0012—Oxidoreductases (1.) acting on nitrogen containing compounds as donors (1.4, 1.5, 1.6, 1.7)
- C12N9/0014—Oxidoreductases (1.) acting on nitrogen containing compounds as donors (1.4, 1.5, 1.6, 1.7) acting on the CH-NH2 group of donors (1.4)
- C12N9/0022—Oxidoreductases (1.) acting on nitrogen containing compounds as donors (1.4, 1.5, 1.6, 1.7) acting on the CH-NH2 group of donors (1.4) with oxygen as acceptor (1.4.3)
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/7088—Compounds having three or more nucleosides or nucleotides
-
- 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
- A61K39/395—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum
- A61K39/39533—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum against materials from animals
- A61K39/3955—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum against materials from animals against proteinaceous materials, e.g. enzymes, hormones, lymphokines
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
- A61P35/02—Antineoplastic agents specific for leukemia
-
- 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
- C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
- C12N15/113—Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing
- C12N15/1137—Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing against enzymes
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/26—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving oxidoreductase
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
- C12Q1/6883—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material
- C12Q1/6886—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material for cancer
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y104/00—Oxidoreductases acting on the CH-NH2 group of donors (1.4)
- C12Y104/03—Oxidoreductases acting on the CH-NH2 group of donors (1.4) with oxygen as acceptor (1.4.3)
- C12Y104/03002—L-Amino-acid oxidase (1.4.3.2)
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/573—Immunoassay; Biospecific binding assay; Materials therefor for enzymes or isoenzymes
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
- G01N33/57407—Specifically defined cancers
- G01N33/57423—Specifically defined cancers of lung
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
- G01N33/57484—Immunoassay; Biospecific binding assay; Materials therefor for cancer involving compounds serving as markers for tumor, cancer, neoplasia, e.g. cellular determinants, receptors, heat shock/stress proteins, A-protein, oligosaccharides, metabolites
-
- 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
- C12N2310/00—Structure or type of the nucleic acid
- C12N2310/10—Type of nucleic acid
- C12N2310/14—Type of nucleic acid interfering N.A.
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q2600/00—Oligonucleotides characterized by their use
- C12Q2600/118—Prognosis of disease development
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q2600/00—Oligonucleotides characterized by their use
- C12Q2600/136—Screening for pharmacological compounds
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q2600/00—Oligonucleotides characterized by their use
- C12Q2600/158—Expression markers
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/90—Enzymes; Proenzymes
- G01N2333/902—Oxidoreductases (1.)
- G01N2333/906—Oxidoreductases (1.) acting on nitrogen containing compounds as donors (1.4, 1.5, 1.7)
- G01N2333/90605—Oxidoreductases (1.) acting on nitrogen containing compounds as donors (1.4, 1.5, 1.7) acting on the CH-NH2 group of donors (1.4)
- G01N2333/90633—Oxidoreductases (1.) acting on nitrogen containing compounds as donors (1.4, 1.5, 1.7) acting on the CH-NH2 group of donors (1.4) with oxygen as acceptor (1.4.3) in general
- G01N2333/90638—Oxidoreductases (1.) acting on nitrogen containing compounds as donors (1.4, 1.5, 1.7) acting on the CH-NH2 group of donors (1.4) with oxygen as acceptor (1.4.3) in general with a definite EC number (1.4.3.-)
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2500/00—Screening for compounds of potential therapeutic value
- G01N2500/04—Screening involving studying the effect of compounds C directly on molecule A (e.g. C are potential ligands for a receptor A, or potential substrates for an enzyme A)
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Immunology (AREA)
- Molecular Biology (AREA)
- Biomedical Technology (AREA)
- Genetics & Genomics (AREA)
- Medicinal Chemistry (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Microbiology (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biotechnology (AREA)
- Hematology (AREA)
- General Engineering & Computer Science (AREA)
- Urology & Nephrology (AREA)
- Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Pharmacology & Pharmacy (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- Pathology (AREA)
- Animal Behavior & Ethology (AREA)
- Cell Biology (AREA)
- Oncology (AREA)
- Food Science & Technology (AREA)
- Epidemiology (AREA)
- General Physics & Mathematics (AREA)
- Hospice & Palliative Care (AREA)
- Biophysics (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Mycology (AREA)
- Endocrinology (AREA)
Abstract
本发明提供了一种肺癌的新型代谢免疫靶点IL4I1及其应用，涉及生物医药技术领域。本发明所述IL4I1蛋白主要分布于肺腺癌肿瘤微环境内的巨噬细胞、树突状细胞及骨髓来源的抑制性细胞中，呈点状聚集。IL4I1的表达水平随肺腺癌肿瘤进展而上调，mRNA、蛋白水平的高表达均提示预后不良。本发明表明IL4I1分子通过代谢色氨酸、调控AHR信号通路促进免疫抑制性肿瘤微环境的形成、促进癌细胞的侵袭和迁移。本发明首次发现了在早期肺腺癌肿瘤微环境内存在高表达的IL4I1，同时首次发现了早期肺腺癌的新型代谢免疫靶点IL4I1，为肿瘤治疗提供了新的药物靶点、新的思路和策略。
Description
技术领域
本发明属于生物医药技术领域，具体涉及一种肺癌的新型代谢免疫靶点IL4I1及其应用。
背景技术
在适应性免疫应答及肿瘤免疫中，T淋巴细胞响应激活的高代谢需求使其非常容易受到生化环境改变的影响，如高度依赖外源性摄取的必需氨基酸以满足其快速增殖及激活的需要。在肿瘤微环境中，髓系来源的细胞如巨噬细胞等能够通过产生分解代谢必需或半必需氨基酸的免疫抑制酶，一方面造成营养缺陷、改变营养物质的可用性，另一方面产生有毒分解代谢物、促进免疫细胞凋亡，从而参与破坏局部抗肿瘤免疫反应。
L-氨基酸氧化酶是在从细菌到哺乳动物的整个进化过程中均有发现的黄素腺嘌呤二核苷酸(FAD)结合酶，通常具有抗感染特性，在调节淋巴细胞过度活化和炎症方面有较为突出的功能。这些酶可根据其氨基酸底物分为：分解代谢半必需氨基酸精氨酸的iNOS(诱导型一氧化氮合酶)和Arg1/Arg2(1/2型精氨酸酶)、分解代谢必需氨基酸色氨酸的IDO1/IDO2(1/2型吲哚胺2,3-双加氧酶)和TDO(色氨酸2,3双加氧酶)、以及分解代谢苯丙氨酸的IL4I1(白细胞介素4诱导的基因1)。
IL4I1基因最早于细胞因子白细胞介素4刺激后的脾B细胞中被发现，并因此得名。其人类基因位于第19号染色体上的白细胞受体复合物内，小鼠基因则位于第7号染色体上，该区域缺陷易患系统性红斑狼疮。人IL4I1主要在淋巴组织、精子、中枢神经系统的稀有细胞中表达。
IL4I1蛋白是一种分泌型的L-苯丙氨酸氧化酶，有糖基化修饰，属于FAD结合酶的L-氨基酸氧化酶(LAAO)家族，既往研究认为，IL4I1能够将苯丙氨酸氧化脱氨成苯丙酮酸，并在此过程中释放H2O2和NH3，对色氨酸和精氨酸也有较低的分解代谢活性。有研究表明，一些苯丙氨酸衍生物可以初步抑制IL4I1的活性，但其特异性有限、且活性相对较低(微摩尔级别)。
在病理状态下，IL4I1主要表达于免疫细胞，尤其是炎症和T辅助细胞1(Th1)刺激状态(Toll样受体、IL1、IFN1/2的配体)的骨髓来源细胞(单核细胞/巨噬细胞和树突细胞)，这种诱导表达诱导依赖于NFκB和STAT1的激活。IL4I1在结节病和结核病患者的慢性Th1肉芽肿中的树突细胞和巨噬细胞群中有显著高表达，在肿瘤浸润巨噬细胞中也有表达。此外，IL4I1在受IL4和CD40L激活的人外周血B细胞中，Th17细胞和CD4+T细胞中有轻微表达，后者的转录受RORT控制。近期研究表明，IL4I1还可由粘膜相关不变T细胞(Mucosal-AssociatedInvariant T cells，MAIT)表达，后者是一种MR1限制性抗菌T细胞亚群，主要在屏障位点以及肝脏和血液中被检测到。常规CD8+T细胞及NK细胞中均未检测到IL4I1的表达。
目前尚未有研究对IL4I1在肿瘤中的蛋白表达情况进行大规模分析，或对其作为肿瘤治疗靶点、与免疫疗法联用的可能性进行探究。
发明内容
有鉴于此，本发明的目的在于提供一种肺癌的新型代谢免疫靶点IL4I1及其应用，为肺腺癌及其他相关癌症提供了新的诊断、治疗方法、联合治疗策略和药物筛选平台。
为了实现上述发明目的，本发明提供以下技术方案：
本发明提供了一种检测IL4I1的试剂在制备肺腺癌诊断工具中的应用，所述IL4I1的氨基酸序列包括SEQ ID NO.1所示的氨基酸序列，或对SEQ ID NO.1所示的氨基酸序列进行取代、缺失和/或添加后形成的与SEQ ID NO.1功能相同的氨基酸序列。
优选的，所述检测IL4I1的试剂包括：血浆蛋白标志物、病理组化标志物、引物或探针、选自IL4I1的多核苷酸序列的外周血ctDNA/cfDNA标志物中的一种或多种的组合。
本发明还提供了IL4I1作为药物作用靶点在筛选治疗肿瘤药物中的应用，所述IL4I1的氨基酸序列包括SEQ ID NO.1所示的氨基酸序列，或对SEQ ID NO.1所示的氨基酸序列进行取代、缺失和/或添加后形成的与SEQ ID NO.1功能相同的氨基酸序列。
优选的，所述药物包括a)～f)中的至少一种功能：
a)治疗肿瘤或者抑制肿瘤生长；
b)抑制肿瘤侵袭和/或转移；
c)促进肿瘤患者的抗肿瘤免疫反应
d)抑制肿瘤微环境中髓系来源的抑制性细胞聚集；
e)促进T细胞增殖与活化，和/或减少T细胞凋亡，和/或增强T细胞效应功能和肿瘤杀伤活性；
f)与肿瘤免疫疗法联用，增强免疫疗法效果。
优选的，所述肿瘤包括肺腺癌、黑色素瘤、慢性淋巴细胞白血病中的一种或多种。
本发明还提供了一种干扰IL4I1表达的试剂，所述试剂包括：干扰IL4I1表达的RNA分子、抗IL4I1抗体、IL4I1小分子抑制剂、IL4I1可溶性蛋白和/或靶向IL4I1的PROTAC降解剂；
所述IL4I1的氨基酸序列包括SEQ ID NO.1所示的氨基酸序列，或对SEQ ID NO.1所示的氨基酸序列进行取代、缺失和/或添加后形成的与SEQ ID NO.1功能相同的氨基酸序列。
优选的，所述干扰IL4I1表达的RNA分子包括①或②中的序列：
①SEQ ID NO.2和SEQ ID NO.3所示的shRNA；
②将SEQ ID NO.2和SEQ ID NO.3所示的shRNA删除或增加或改变一个或几个核苷酸，且与SEQ ID NO.2和SEQ ID NO.3所示的shRNA保持相同功能的核苷酸。
本发明还提供了一种治疗肺腺癌的药物，所述药物包括上述的试剂。
本发明还提供了IL4I1作为靶点在开发或设计肿瘤免疫治疗药物、肿瘤联合治疗策略中的应用，所述IL4I1的氨基酸序列包括SEQ ID NO.1所示的氨基酸序列，或对SEQ IDNO.1所示的氨基酸序列进行取代、缺失和/或添加后形成的与SEQ ID NO.1功能相同的氨基酸序列。
有益效果：本发明提供了一种检测IL4I1的试剂在制备肺腺癌诊断工具中的应用，所述IL4I1蛋白主要分布于肺腺癌肿瘤微环境内的巨噬细胞、树突状细胞及骨髓来源的抑制性细胞中，呈点状聚集。IL4I1的表达水平随肺腺癌肿瘤进展而上调，mRNA、蛋白水平的高表达均提示预后不良。在本发明实施例中，构建IL4I1特异性敲除细胞系，发现与野生型对照组相比，该IL4I1特异性敲除细胞系代谢色氨酸能力显著降低，且下游AHR信号通路相关分子表达水平显著下降，癌细胞侵袭迁移能力显著降低，表明IL4I1分子通过代谢色氨酸、调控AHR信号通路促进免疫抑制性肿瘤微环境的形成、促进癌细胞的侵袭和迁移。参与调节肺腺癌发生发展过程中肿瘤微环境内的代谢及免疫功能。本发明首次发现了在早期肺腺癌肿瘤微环境内存在高表达的IL4I1，同时首次发现了早期肺腺癌的新型代谢免疫靶点IL4I1，为肿瘤治疗提供了新的药物靶点、新的思路和策略。
附图说明
图1为非靶向蛋白质组学检测早期肺腺癌不同阶段临床样本的癌组织及癌旁组织中IL4I1的表达情况；
图2为单细胞转录组学技术明确早期肺腺癌肿瘤微环境内主要表达IL4I1的细胞类型；
图3为单细胞转录组学技术明确早期肺腺癌不同阶段IL4I1的表达情况变化；
图4为通过免疫组化技术，在早期肺腺癌石蜡切片中识别内源性IL4I1的蛋白并验证其表达情况；
图5为通过免疫组化技术，在早期肺腺癌石蜡切片中识别内源性IL4I1的细胞亚型分布；
图6为IL4I1mRNA水平统计；
图7为IL4I1水平统计及ROC曲线；
图8为在A549细胞中过表达C端GFP标签IL4I1的免疫荧光结果及亚细胞定位；
图9为IL4I1过表达及敲低细胞系的构建验证；
图10为IL4I1过表达及敲低对AHR信号通路的影响；
图11为IL4I1过表达及敲低对细胞迁移侵袭能力的影响。
具体实施方式
本发明提供了一种检测IL4I1的试剂在制备肺腺癌诊断工具中的应用，所述IL4I1的氨基酸序列包括SEQ ID NO.1所示的氨基酸序列，或对SEQ ID NO.1所示的氨基酸序列进行取代、缺失和/或添加后形成的与SEQ ID NO.1功能相同的氨基酸序列。
本发明SEQ ID NO.1所示的氨基酸序列优选为：MAPLALHLLVLVPILLSLVASQDWKAERSQDPFEKCMQDPDYEQLLKVVTWGLNRTLKPQRVIVVGAGVAGLVAAKVLSDAGHKVTILEADNRIGGRIFTYRDQNTGWIGELGAMRMPSSHRILHKLCQGLGLNLTKFTQYDKNTWTEVHEVKLRNYVVEKVPEKLGYALRPQEKGHSPEDIYQMALNQALKDLKALGCRKAMKKFERHTLLEYLLGEGNLSRPAVQLLGDVMSEDGFFYLSFAEALRAHSCLSDRLQYSRIVGGWDLLPRALLSSLSGLVLLNAPVVAMTQGPHDVHVQIETSPPARNLKVLKADVVLLTASGPAVKRITFSPPLPRHMQEALRRLHYVPATKVFLSFRPFWREEHIEGGHSNTDRPSRMIFYPPPREGALLLASYTWSDAAAAFAGLSREEALRLALDDVAALHGPVVRQLWDGTGVVKRWAEDQHSQGGFVVQPPALWQTEKDDWTVPYGRIYFAGEHTAYPHGWVETAVKSALRAAIKINSRKGPASDTASPEGHASDMEGQGHVHGVASSPSHDLAKEEGSHPPVQGQLSLQNTTHTRTSH。
在本发明中，所述IL4I1蛋白主要分布于肺腺癌肿瘤微环境内的巨噬细胞、树突状细胞及骨髓来源的抑制性细胞中，呈点状聚集；IL4I1的表达水平随肺腺癌肿瘤进展而上调，mRNA、蛋白水平的高表达均提示预后不良，可作为检测和/或治疗肺腺癌的生物标志物。
本发明所述检测IL4I1的试剂，优选包括：血浆蛋白标志物、病理组化标志物、引物或探针、选自IL4I1的多核苷酸序列的外周血ctDNA/cfDNA标志物中的一种或多种的组合。
本发明还提供了IL4I1作为药物作用靶点在筛选治疗肿瘤药物中的应用，所述IL4I1的氨基酸序列包括SEQ ID NO.1所示的氨基酸序列，或对SEQ ID NO.1所示的氨基酸序列进行取代、缺失和/或添加后形成的与SEQ ID NO.1功能相同的氨基酸序列。
在本发明实施例中，通过构建IL4I1特异性敲除细胞系，发现与野生型对照组相比，该IL4I1特异性敲除细胞系代谢色氨酸能力显著降低，且下游AHR信号通路相关分子表达水平显著下降，癌细胞侵袭迁移能力显著降低，表明IL4I1分子通过代谢色氨酸、调控AHR信号通路促进免疫抑制性肿瘤微环境的形成、促进癌细胞的侵袭和迁移；参与调节肺腺癌发生发展过程中肿瘤微环境内的代谢及免疫功能，因此可将IL4I1作为药物作用靶点，用于筛选治疗肿瘤药物。本发明所述药物优选包括a)～f)中的至少一种功能：
a)治疗肿瘤或者抑制肿瘤生长；
b)抑制肿瘤侵袭和/或转移；
c)促进肿瘤患者的抗肿瘤免疫反应
d)抑制肿瘤微环境中髓系来源的抑制性细胞聚集；
e)促进T细胞增殖与活化，和/或减少T细胞凋亡，和/或增强T细胞效应功能和肿瘤杀伤活性；
f)与肿瘤免疫疗法联用，增强免疫疗法效果。
在本发明中，所述肿瘤优选包括肺腺癌、黑色素瘤、慢性淋巴细胞白血病中的一种或多种，更优选为肺腺癌。
本发明还提供了一种干扰IL4I1表达的试剂，所述试剂包括：干扰IL4I1表达的RNA分子、抗IL4I1抗体、IL4I1小分子抑制剂、IL4I1可溶性蛋白和/或靶向IL4I1的PROTAC降解剂；
所述IL4I1的氨基酸序列包括SEQ ID NO.1所示的氨基酸序列，或对SEQ ID NO.1所示的氨基酸序列进行取代、缺失和/或添加后形成的与SEQ ID NO.1功能相同的氨基酸序列。
本发明所述试剂优选为干扰IL4I1表达的RNA分子，且所述干扰IL4I1表达的RNA分子优选包括①或②中的序列：
①SEQ ID NO.2和SEQ ID NO.3所示的shRNA；
②将SEQ ID NO.2和SEQ ID NO.3所示的shRNA删除或增加或改变一个或几个核苷酸，且与SEQ ID NO.2和SEQ ID NO.3所示的shRNA保持相同功能的核苷酸。
本发明所述shRNA序列能够与IL4I1的CDS区结合以干扰mRNA翻译，具体序列如下：
正向引物(SEQ ID NO.2)：CCGGaggcacacgctcttggaatatCTCGAGatattccaagagcgtgtgcctTTTTTG；
反向引物(SEQ ID NO.3)：aattCAAAAAaggcacacgctcttggaatatCTCGAGatattccaagagcgtgtgcct。利用本发明所述shRNA可敲低IL4I1的表达，实施例证实，敲低IL4I1的表达后对AHR信号通路下游分子的表达有显著抑制效果，提示在肺腺癌中IL4I1表达的增强可能激活AHR信号通路；IL4I1敲低的肿瘤细胞迁移能力明显下降，提示IL4I1能够促进肿瘤细胞迁移；同时还利用超表达IL4I1获得了同样能够得出上述结论的结果，因此可将其用于治疗肺腺癌。
本发明还提供了一种治疗肺腺癌的药物，所述药物包括上述的试剂。
本发明对所述药物的剂型以及制备方法并没有特殊限定，利用本领域的常规剂型，根据常规制备方法添加药学上可接受的辅料即可。
本发明还提供了IL4I1作为靶点在开发或设计肿瘤免疫治疗药物、肿瘤联合治疗策略中的应用，所述IL4I1的氨基酸序列包括SEQ ID NO.1所示的氨基酸序列，或对SEQ IDNO.1所示的氨基酸序列进行取代、缺失和/或添加后形成的与SEQ ID NO.1功能相同的氨基酸序列。本发明所述应用优选与上述相同，在此不再赘述。
下面结合实施例对本发明提供的一种肺癌的新型代谢免疫靶点IL4I1及其应用进行详细的说明，但是不能把它们理解为对本发明保护范围的限定。
实施例1
1、通过Bulk非靶向蛋白质组学技术大规模鉴定IL4I1在早期肺腺癌不同阶段临床样本中的蛋白水平表达情况：
①蛋白质组学样本前处理：
选取CT影像学表现为磨玻璃结节及实性肺腺癌、术后病理诊断为腺癌的亚实性结节及实性结节患者各48例，取手术切除的癌灶及正常癌旁组织。每100mg组织使用1mL组织裂解液在冰浴条件下的组织匀浆仪中充分破碎，离心弃去组织残渣后，测定上清蛋白浓度，每样本取100μg蛋白上清进行后续处理。蛋白上清经尿素变性、还原烷基化处理后，于37℃孵箱中胰酶酶解过夜。将酶解后的样本经10Kd超滤管超滤离心，滤液脱盐处理后，真空干燥，-80℃保存至上机；
②长梯度液相色谱分离联合高分辨率轨道阱质谱仪检测：
取-80℃保存的蛋白样本，每样本取6μl0.1％甲酸水溶液进行复溶混匀，采用120分钟长梯度、纳升级液相色谱进行梯度分离，而后经超高静电场轨道阱傅里叶变换台式质谱仪Thermo Q Exactive-HF对肽段进行快速的识别、表征、定量和确认。
质谱具体参数如下：
首先通过长梯度、高pH值液相色谱(HPLC)对酶解后的肽段进行分级，以降低样品复杂性。将冷冻干燥的酶解后肽段溶解于缓冲液A(2％乙腈，pH9.5)中，上样至XbridgeC18色谱柱(4.6mm×100mm，3.5μm)，以0.6ml/min的流速逐步从0过渡到95％缓冲液B(98％ACN，pH9.5)，梯度洗脱60分钟，将收集的等分试样合并为10份。
而后在与Orbitrap-QExactive HF质谱联用的EASY-nLC1000LC系统上分析肽样品。将肽重新溶解于流动相A(2％ACN和0.1％甲酸)中，上样到填充有C18树脂(粒径3μm，孔径)的18cm长的自制C18纳米毛细管分析柱上(柱内径75mm)。以350nL/min的恒定流速将肽分离，梯度120分钟(缓冲液A：0.1％甲酸溶于去离子水，缓冲液B：0.1％甲酸溶于90％ACN中)。
其中0-50分钟为5到45％的缓冲液B，50-55分钟为45％至80％的缓冲液B，55-60分钟为80％的缓冲液B。
洗脱的肽在2k伏电压下离子化并引入质谱，质谱在数据依赖的采集模式下操作。对于MS1全扫描，Orbitrap质量分析器以120,000的高分辨率采集m/z范围为350至1300的离子，自动增益控制(AGC)设置为5×105，最大离子注入时间为50ms。MS2采集以最高速度模式进行，占空比时间(duty cycle time)为3秒。MS2选择母离子并以更高能量的碰撞解离(HCD)对母离子进行碎片化，归一化碰撞能量为32％，碎片离子采用离子阱质量分析仪进行分析，AGC设置为7000。MS2的最大离子注入时间为35ms，动态排除为60s。
采用293T细胞的全细胞蛋白提取液作为批内质控，每10个样本后加入一个质控样本以保证数据的可重复性。
使用支持Andromeda搜索引擎的MaxQuant(版本1.6.5.0)软件通过UniProt人类蛋白质组数据库(版本2019-03-07，20,404个序列)搜索质谱下机生成的MS原始文件。蛋白酶选择胰蛋白酶(Trypsin/P)，最多允许2个遗漏的酶切位点，氨基甲酸酯(C)设定为固定修饰，氧化(M)和乙酰化(N端)设定为可变修饰，蛋白和肽段的错误发现率(FDR)均设置为小于1％。
使用X！Tandem(2017.2.14版本)在具有诱饵序列的患者衍生蛋白质数据库中搜索质谱下机生成的MS/MS数据。对于变异肽鉴定，设定FDR小于0.1％以保证变异肽与野生型之间没有明显的偏差。通过ANNOVARCOSMIC数据库对变异肽进行注释，使用NIPS评估单个氨基酸变异的有害风险。
下机数据通过Uniprot数据库比对处理后，采用IL4I1峰图中的曲线下面积(LFQintensity)对不同组别中的IL4I1表达水平进行比较；
③非靶向蛋白质组学质谱结果显示，IL4I1在早期肺腺癌中的表达水平远高于正常癌旁组织，且随癌症进展呈现表达上调的趋势(见图1中B，病人病理特征见表1)。
表1入组患者临床信息表
2、通过单细胞转录组学技术明确IL4I1在早期肺腺癌不同阶段的mRNA表达情况及主要来源细胞，具体方法如下：
①利用酶解消化方法分离得到肺癌组织和正常对照组织的单细胞悬液，流式分选得到存活状态的上皮、免疫和间质细胞，将各组分混合后上机构建10XGenomics单细胞转录组文库，利用Illumina平台进行高通量二代测序，使用CellRanger对样本的原始数据进行拆分、基因组比对、过滤低质量数据、处理条形码，最终获得基因-细胞矩阵，对细胞进行降维、聚类，得到细胞亚群；
②对聚类后的细胞进行Marker基因提取并鉴定细胞类型，利用InferCNVR包分析各上皮细胞群的基因拷贝变异情况，鉴定肿瘤细胞(见图2中A)；
③提取IL4I1在不同细胞类型中的mRNA表达水平并绘制UMAP图，结果显示IL4I1主要表达于髓系来源的细胞——即树突状细胞与巨噬细胞中，少量表达于部分T细胞中(见图2中B)；
④对不同阶段肺腺癌患者的单细胞转录组学测序结果中IL4I1的mRNA表达水平进行比较，IL4I1的mRNA表达集中于单核细胞中、且随肺腺癌的进展而增强(见图3)，证实IL4I1在mRNA及蛋白水平均随肺腺癌的进展而增强，且其主要通过髓系来源的单核-巨噬细胞表达。
3、通过免疫组化技术，在早期肺腺癌石蜡切片中识别内源性IL4I1的蛋白以验证其表达情况及细胞亚型分布：
①临床手术切除肿瘤组织用无菌纱布吸干血液后立即投入处于液氮的异戊烷中速冻，置于-80℃中保存，后利用石蜡包埋剂对组织进行包埋和制作连续切片(切片厚度2mm)，选取合适组织切面进行HE染色获得组织学背景信息；
②使用IL4I1特异性兔源抗体(兔抗人IL4I1抗体，222102，Abcam公司)对石蜡切片进行免疫组化检测，采用AxioScanZ1扫片仪进行全片扫描，比较不同阶段癌组织的IL4I1表达情况。结果如图4所示，内源性IL4I1在正常肺组织中无表达，在亚实性结节阶段的肺腺癌及实性肺腺癌中呈团块状聚集表达、且主要表达于细胞较小、核圆的免疫细胞中。实性肺腺癌中的内源性IL4I1表达情况明显高于实性结节；
③使用单核细胞表面抗原CD68的特异性兔源抗体对相同组织的连续切片进行免疫组化染色，采用AxioScanZ1扫片仪进行全片扫描，将同一组织连续切片中IL4I1的免疫组化结果与CD68染色结果进行整合，以观察两者共定位情况。结果如图5所示，IL4I1阳性细胞与CD68阳性细胞有明显共定位，提示在肺腺癌中，IL4I1确由CD68阳性的巨噬细胞表达。
实施例2
1、关于IL4I1mRNA作为分子标记诊断标志物在早期肺腺癌检测中的应用：
①肺腺癌组织的采集、处理与总RNA提取：
采集CT影像及病理学诊断为肺腺癌的患者术中切除癌组织及其配对正常癌旁组织，每样本取100mg组织，加入1mlTrizol后在组织匀浆仪中充分研磨成粉末，提取组织总RNA；
②每样本取2μgRNA，使用带polyT的随机引物，按照5xALLINONE试剂盒(ABM公司)说明书进行cDNA反转录，产物立即用于qPCR检测，或于-80℃保存至使用；
③qPCR检测：
根据NCBI中IL4I1mRNA序列、依据跨外显子原则设计并优化针对IL4ICDS区的引物，由北京睿博测序公司合成；利用qPCR对20对肺腺癌样本及其配对癌旁组织的cDNA进行IL4I1表达水平检测，引物序列如下：
正向引物：CGCCCGAAGACATCTACCAG(SEQ ID NO.4)；
反向引物：GATATTCCAAGAGCGTGTGCC(SEQ ID NO.5)；
内参正向引物：CACCATTGGCAATGAGCGGTTC(SEQ ID NO.6)；
内参反向引物：AGGTCTTTGCGGATGTCCACGT(SEQ ID NO.7)。
体系(20μl)：模板2μl、引物F+R(10μM)0.4μl、2xperfectstart qPCRsupermix10μl、PassivereferenceDye0.4μl和余量的双蒸水；
程序：94℃30s；94℃5s，60℃34s，40个循环；94℃30s，65℃5s。
④按照上述条件，用qPCR方法检测早期肺腺癌组织中的IL4I1表达情况，结果发现与癌旁组织相比，实性肺腺癌中IL4I1mRNA含量明显升高，见图6；绘制ROC曲线，其曲线下面积(AUC)在0.99以上(图7)，提示IL4I1mRNA可作为肺腺癌的分子病理诊断标志物，使用IL4I1mRNA可作为肺腺癌在分子病理诊断方面必要和有益的补充。
2、IL4I1蛋白作为分子病理诊断标志物在早期肺腺癌检测中的应用
①肺腺癌组织石蜡切片的采集：
采集CT影像及病理学诊断为肺腺癌的患者术中切除癌组织及其配对正常癌旁组织的石蜡包埋样本，制作组织连续切片，切片厚度为2mm，选取合适的组织切面进行HE染色以获得组织学背景信息；
②使用特异性识别内源性IL4I1的兔源抗体(Abcam公司，货号AB222102)进行组织染色，采用AxioScanZ1扫片仪进行全片扫描，整合HE染色结果并通过ImageJ软件对IL4I1表达情况按染色面积比例进行打分；
③按照上述条件，用免疫组化方法检测早期肺腺癌组织的IL4I1蛋白表达情况，结果发现与癌旁组织、亚实性结节阶段的癌组织相比，实性腺癌中IL4I1的表达水平明显高于亚实性结节阶段及正常癌旁组织。
实施例3
1、真核外源过表达IL4I1载体的构建及在A549细胞中观察IL4I1的定位，具体方法如下：
①将人肺腺癌cDNA进行PCR扩增得到编码IL4I1的核苷酸序列，通过同源重组插入C端带有GFP标签的真核过表达载体pEGFP-C1中(图8)，通过DH5α感受态进行质粒扩增，并将构建成功的质粒载体送北京睿博公司测序，测序结果经比对正确后使用；
PCR扩增所用引物：
pEGFP-C3-IL4I1-F(SEQ ID NO.30)：taccggtcgccaccatggccccattggcc
pEGFP-C3-IL4I1-R(SEQ ID NO.31)：gcccttgctcaccatatgcgaggtcctcgtg
PCR反应程序：95℃预变性5min；95℃变性20s，65℃退火20s，72℃延伸1min，30循环；72℃再延伸5min。
荧光实时定量PCR反应体系(20μl)：cDNA(0.1μg/μl)1μl、正向引物(10μm)0.4μl、反向引物(10μm)0.4μl、2×TOPGreenqPCRMix10μl、5×ReferenceDye0.4μl、ddH2O7.8μl。
实时荧光定量PCR反应条件：95℃30s；95℃30s，40循环；60℃34s，40循环。
②在A549细胞中转染IL4I1-GFP质粒，同时用DAPI标记细胞核，在共聚焦显微镜下观察，发现GFP标记的IL4I1在整个胞浆内呈弥散样分布，表明其主要为分泌性的胞浆蛋白(见图8)。
2、构建IL4I1过表达及敲低细胞系：
①通过RNAi技术构建IL4I1敲低细胞系：
RNAi由包含shRNA的质粒稳定表达。设计能够与IL4I1的CDS区结合以干扰mRNA翻译的RNAi序列，由北京睿博测序公司合成SEQ ID NO.2和SEQ ID NO.3所示的引物；
取浓度为10μM的正反向引物各25μL于0.5mLEP管中，95℃水浴10min后缓慢自然降至室温冷却，得到双链引物序列；
使用EcoRI-HF和AgeI-HF限制性内切酶于37℃酶切pLKO-TRC质粒，1.2％琼脂糖凝胶电泳回收酶切后的线性质粒骨架，使用T4DNA连接酶将双链shRNA引物序列插入pLKO质粒骨架中，通过Stbl3感受态对插入成功的阳性质粒进行扩增。构建成功的质粒送北京睿博测序公司测序，结果经比对正确后使用；
将pLKO-shIL4I1载体与工具质粒pMD2.G、psPAX共转染293T细胞，在10cm2培养皿中汇合度为70％的293T细胞中转染，72小时后收集培养上清，72000g离心2小时后，弃去上清并用100μLDMEM培养基重悬沉淀，测定病毒滴度；
共转染体系：pLKO-shIL4I14μg、pMD2.G1μg、psPAX3μg、PEI2倍质粒体积和optiMEM250μl。
向10cm2培养皿中汇合度为70％的A549、H1299、THP-1细胞中加入50μL病毒浓缩液，感染48h后换新鲜培养基并加入puromycin以筛选阳性细胞。筛选五天后，得到转入质粒的阳性细胞，将阳性细胞计数，并按照每个96孔板种50个细胞的密度进行铺板，共计种10个96孔板。培养14～20天后，通过显微镜观察，可见在部分孔中的阳性细胞单克隆。挑取单克隆至12孔板的一孔中继续培养，待长满后取部分细胞悬液，通过免疫印迹实验验证IL4I1表达情况；
②过表达IL4I1细胞系的构建：
利用与上相同的方法，将人肺腺癌cDNA进行PCR扩增得到编码IL4I1的核苷酸序列，通过同源重组插入质粒pcDNA3.1中(图9)，构建真核过表达载体pcDNA3.1-IL4I1，质粒经北京睿博测序公司测序验证后使用。使用polyplus公司的jetPRIME试剂，向在6孔板中培养的汇合度为70％的A549、H1299、THP-1细胞转染上述载体8μg，48小时候收集细胞，通过免疫印迹实验确认IL4I1过表达情况，见图9；
3、IL4I1后AHR信号通路下游分子有明显变化：
分别收取10cm2培养皿的IL4I1过表达及敲低细胞，使用Trizol提取总RNA并反转录为cDNA，通过qPCR对AHR信号通路下游分子的表达情况进行检测，所用引物见表2。
表2 AHR信号通路下游分子qPCR引物
②结果显示，过表达IL4I1能够显著激活AHR信号通路下游分子的表达，而敲低IL4I1则对AHR信号通路下游分子的表达有显著抑制效果(图10)，提示在肺腺癌中IL4I1表达的增强可能激活AHR信号通路。
4、IL4I1过表达能够促进肿瘤细胞迁移，其具体验证方法如下：
①通过细胞划痕实验明确IL4I1过表达能够促进肿瘤细胞迁移，包括以下步骤：
在6孔板中分别接种2×105个过表达IL4I1、敲低IL4I1及野生型的A549细胞，加2mLDMEM培养过夜至汇合度为90％时，用干净灭菌的枪头在6孔板内划线，每孔3条纵线、3条横线；
划线后换新鲜培养基继续培养，分别于0h、24h、72h拍照，使用ImageJ处理图像并计算woundclosure比例；
结果如图11所示，IL4I1敲低的肿瘤细胞迁移能力明显下降，而IL4I1过表达的肿瘤细胞迁移能力则明显上升，提示IL4I1能够促进肿瘤细胞迁移。
②通过transwell实验进一步验证IL4I1过表达能够促进肿瘤细胞迁移，包括以下步骤：
向24孔板中分别种入105个过表达IL4I1、敲低IL4I1及野生型的THP-1细胞，向上层小室中种入104个A549细胞，培养过夜后使用结晶紫固定染色，计算穿孔细胞比率；
结果表明，IL4I1过表达的THP-1细胞能够促进A549细胞向下层小室迁移，而敲低IL4I1的THP-1细胞则不能，提示IL4I1确实能够促进肿瘤细胞迁移。
以上所述仅是本发明的优选实施方式，应当指出，对于本技术领域的普通技术人员来说，在不脱离本发明原理的前提下，还可以做出若干改进和润饰，这些改进和润饰也应视为本发明的保护范围。
序列表
<110> 北京大学
北京大学人民医院
<120> 一种肺癌的新型代谢免疫靶点IL4I1及其应用
<160> 31
<170> SIPOSequenceListing 1.0
<210> 1
<211> 566
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 1
Met Ala Pro Leu Ala Leu His Leu Leu Val Leu Val Pro Ile Leu Leu
1 5 10 15
Ser Leu Val Ala Ser Gln Asp Trp Lys Ala Glu Arg Ser Gln Asp Pro
20 25 30
Phe Glu Lys Cys Met Gln Asp Pro Asp Tyr Glu Gln Leu Leu Lys Val
35 40 45
Val Thr Trp Gly Leu Asn Arg Thr Leu Lys Pro Gln Arg Val Ile Val
50 55 60
Val Gly Ala Gly Val Ala Gly Leu Val Ala Ala Lys Val Leu Ser Asp
65 70 75 80
Ala Gly His Lys Val Thr Ile Leu Glu Ala Asp Asn Arg Ile Gly Gly
85 90 95
Arg Ile Phe Thr Tyr Arg Asp Gln Asn Thr Gly Trp Ile Gly Glu Leu
100 105 110
Gly Ala Met Arg Met Pro Ser Ser His Arg Ile Leu His Lys Leu Cys
115 120 125
Gln Gly Leu Gly Leu Asn Leu Thr Lys Phe Thr Gln Tyr Asp Lys Asn
130 135 140
Thr Trp Thr Glu Val His Glu Val Lys Leu Arg Asn Tyr Val Val Glu
145 150 155 160
Lys Val Pro Glu Lys Leu Gly Tyr Ala Leu Arg Pro Gln Glu Lys Gly
165 170 175
His Ser Pro Glu Asp Ile Tyr Gln Met Ala Leu Asn Gln Ala Leu Lys
180 185 190
Asp Leu Lys Ala Leu Gly Cys Arg Lys Ala Met Lys Lys Phe Glu Arg
195 200 205
His Thr Leu Leu Glu Tyr Leu Leu Gly Glu Gly Asn Leu Ser Arg Pro
210 215 220
Ala Val Gln Leu Leu Gly Asp Val Met Ser Glu Asp Gly Phe Phe Tyr
225 230 235 240
Leu Ser Phe Ala Glu Ala Leu Arg Ala His Ser Cys Leu Ser Asp Arg
245 250 255
Leu Gln Tyr Ser Arg Ile Val Gly Gly Trp Asp Leu Leu Pro Arg Ala
260 265 270
Leu Leu Ser Ser Leu Ser Gly Leu Val Leu Leu Asn Ala Pro Val Val
275 280 285
Ala Met Thr Gln Gly Pro His Asp Val His Val Gln Ile Glu Thr Ser
290 295 300
Pro Pro Ala Arg Asn Leu Lys Val Leu Lys Ala Asp Val Val Leu Leu
305 310 315 320
Thr Ala Ser Gly Pro Ala Val Lys Arg Ile Thr Phe Ser Pro Pro Leu
325 330 335
Pro Arg His Met Gln Glu Ala Leu Arg Arg Leu His Tyr Val Pro Ala
340 345 350
Thr Lys Val Phe Leu Ser Phe Arg Pro Phe Trp Arg Glu Glu His Ile
355 360 365
Glu Gly Gly His Ser Asn Thr Asp Arg Pro Ser Arg Met Ile Phe Tyr
370 375 380
Pro Pro Pro Arg Glu Gly Ala Leu Leu Leu Ala Ser Tyr Thr Trp Ser
385 390 395 400
Asp Ala Ala Ala Ala Phe Ala Gly Leu Ser Arg Glu Glu Ala Leu Arg
405 410 415
Leu Ala Leu Asp Asp Val Ala Ala Leu His Gly Pro Val Val Arg Gln
420 425 430
Leu Trp Asp Gly Thr Gly Val Val Lys Arg Trp Ala Glu Asp Gln His
435 440 445
Ser Gln Gly Gly Phe Val Val Gln Pro Pro Ala Leu Trp Gln Thr Glu
450 455 460
Lys Asp Asp Trp Thr Val Pro Tyr Gly Arg Ile Tyr Phe Ala Gly Glu
465 470 475 480
His Thr Ala Tyr Pro His Gly Trp Val Glu Thr Ala Val Lys Ser Ala
485 490 495
Leu Arg Ala Ala Ile Lys Ile Asn Ser Arg Lys Gly Pro Ala Ser Asp
500 505 510
Thr Ala Ser Pro Glu Gly His Ala Ser Asp Met Glu Gly Gln Gly His
515 520 525
Val His Gly Val Ala Ser Ser Pro Ser His Asp Leu Ala Lys Glu Glu
530 535 540
Gly Ser His Pro Pro Val Gln Gly Gln Leu Ser Leu Gln Asn Thr Thr
545 550 555 560
His Thr Arg Thr Ser His
565
<210> 2
<211> 58
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 2
ccggaggcac acgctcttgg aatatctcga gatattccaa gagcgtgtgc cttttttg 58
<210> 3
<211> 58
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 3
aattcaaaaa aggcacacgc tcttggaata tctcgagata ttccaagagc gtgtgcct 58
<210> 4
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 4
cgcccgaaga catctaccag 20
<210> 5
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 5
gatattccaa gagcgtgtgc c 21
<210> 6
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 6
caccattggc aatgagcggt tc 22
<210> 7
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 7
aggtctttgc ggatgtccac gt 22
<210> 8
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 8
attgtgccga gtcccatatc 20
<210> 9
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 9
aagcaggcgt gcattagact 20
<210> 10
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 10
ttccacgata tggatttacg g 21
<210> 11
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 11
gtttcctgtt gcattgagtc c 21
<210> 12
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 12
gacgccttta tcctctctgc g 21
<210> 13
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 13
acgacctgat ccaattctgc c 21
<210> 14
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 14
gatgtccgta aggtcttgcc 20
<210> 15
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 15
tccagtctcc atcacgaaat 20
<210> 16
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 16
ctcgccagtg aaatgatggc t 21
<210> 17
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 17
gtcggagatt cgtagctgga t 21
<210> 18
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 18
catcaatgac aaggtggcca ag 22
<210> 19
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 19
gggcttgatg gggtgatagg 20
<210> 20
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 20
caaatcctct gggagttgga 20
<210> 21
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 21
gtccaaggct gtcatcgtct 20
<210> 22
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 22
caccctctag caatgtcaac tc 22
<210> 23
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 23
cagactcggg atactctctc c 21
<210> 24
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 24
ctgaccgcag agtcttttcc t 21
<210> 25
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 25
gagtggtttg gctggggtaa 20
<210> 26
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 26
ctgcctgggt ttccatcttc t 21
<210> 27
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 27
gccattcatg tcagagctac act 23
<210> 28
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 28
caccattggc aatgagcggt tc 22
<210> 29
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 29
aggtctttgc ggatgtccac gt 22
<210> 30
<211> 29
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 30
taccggtcgc caccatggcc ccattggcc 29
<210> 31
<211> 31
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 31
gcccttgctc accatatgcg aggtcctcgt g 31
Claims (7)
1.一种检测IL4I1的试剂在制备肺腺癌诊断工具中的应用，其特征在于，所述IL4I1的氨基酸序列如SEQ ID NO.1所示。
2.根据权利要求1所述应用，其特征在于，所述检测IL4I1的试剂包括：血浆蛋白标志物、病理组化标志物、引物或探针、选自IL4I1的多核苷酸序列的外周血ctDNA/cfDNA标志物中的一种或多种的组合。
3.IL4I1作为药物作用靶点在筛选治疗肺腺癌药物中的应用，其特征在于，所述IL4I1的氨基酸序列如SEQ ID NO.1所示。
4.根据权利要求3所述应用，其特征在于，所述药物包括a)～f)中的至少一种功能：
a)治疗肿瘤或者抑制肿瘤生长；
b)抑制肿瘤侵袭和/或转移；
c)促进肿瘤患者的抗肿瘤免疫反应；
d)抑制肿瘤微环境中髓系来源的抑制性细胞聚集；
e)促进T细胞增殖与活化，和/或减少T细胞凋亡，和/或增强T细胞效应功能和肿瘤杀伤活性；
f)与肿瘤免疫疗法联用，增强免疫疗法效果。
5.一种干扰IL4I1表达的试剂，其特征在于，所述试剂为干扰IL4I1表达的RNA分子；
所述IL4I1的氨基酸序列如SEQ ID NO.1所示；
所述干扰IL4I1表达的RNA分子的序列如SEQ ID NO.2和SEQ ID NO.3所示。
6.一种治疗肺腺癌的药物，其特征在于，所述药物包括权利要求5所述的试剂。
7.IL4I1作为靶点在开发或设计肺腺癌免疫治疗药物、肺腺癌联合治疗策略中的应用，其特征在于，所述IL4I1的氨基酸序列如SEQ ID NO.1所示。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210360734.5A CN115044567B (zh) | 2022-04-07 | 2022-04-07 | 一种肺癌的代谢免疫靶点il4i1及其应用 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210360734.5A CN115044567B (zh) | 2022-04-07 | 2022-04-07 | 一种肺癌的代谢免疫靶点il4i1及其应用 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN115044567A CN115044567A (zh) | 2022-09-13 |
| CN115044567B true CN115044567B (zh) | 2023-12-12 |
Family
ID=83157636
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202210360734.5A Active CN115044567B (zh) | 2022-04-07 | 2022-04-07 | 一种肺癌的代谢免疫靶点il4i1及其应用 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN115044567B (zh) |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2013052108A2 (en) * | 2011-10-03 | 2013-04-11 | Oncocyte Corporation | Methods and compositions for the treatment and diagnosis of ovarian cancer |
| CN108795938A (zh) * | 2018-06-21 | 2018-11-13 | 中国科学院北京基因组研究所 | 肺腺癌外泌体特异miRNA及其靶基因与应用 |
| EP3835432A1 (en) * | 2019-12-10 | 2021-06-16 | Deutsches Krebsforschungszentrum, Stiftung des öffentlichen Rechts | Interleukin-4-induced gene 1 (il4i1) and respective metabolites as biomarkers for cancer |
| CN114126634A (zh) * | 2019-04-10 | 2022-03-01 | 德国癌症研究公共权益基金会 | 作为生物标志物的白介素-4诱导基因1（il4i1）及其用途 |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20110152115A1 (en) * | 2003-09-03 | 2011-06-23 | The United States of America, as represented by the Secretary, Department of Health and | Methods for identifying, diagnosing, and predicting survival of lymphomas |
-
2022
- 2022-04-07 CN CN202210360734.5A patent/CN115044567B/zh active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2013052108A2 (en) * | 2011-10-03 | 2013-04-11 | Oncocyte Corporation | Methods and compositions for the treatment and diagnosis of ovarian cancer |
| CN108795938A (zh) * | 2018-06-21 | 2018-11-13 | 中国科学院北京基因组研究所 | 肺腺癌外泌体特异miRNA及其靶基因与应用 |
| CN114126634A (zh) * | 2019-04-10 | 2022-03-01 | 德国癌症研究公共权益基金会 | 作为生物标志物的白介素-4诱导基因1（il4i1）及其用途 |
| EP3835432A1 (en) * | 2019-12-10 | 2021-06-16 | Deutsches Krebsforschungszentrum, Stiftung des öffentlichen Rechts | Interleukin-4-induced gene 1 (il4i1) and respective metabolites as biomarkers for cancer |
Non-Patent Citations (7)
| Title |
|---|
| IL4I1 Is a Metabolic Immune Checkpoint that Activates the AHR and Promotes Tumor Progression;Ahmed Sadik;Cell;第1882卷(第5期);1252-1270 * |
| Q96RQ9.1.RecName: Full=L-amino-acid oxidase * |
| Requirement of Aryl Hydrocarbon Receptor Overexpression for CYP1B1 Up-Regulation and Cell Growth in Human Lung Adenocarcinomas;Jinghua Tsai Chang;HUMAN CANCER BIOLOGY;第13卷(第1期);38-45 * |
| Short=hFIG1 ; Flags: Precursor.UniProtKB/Swiss-Prot.2022,origin部分. * |
| Short=hIL4I1 ; AltName: Full=Protein Fig-1 * |
| Short=IL4-induced protein 1 * |
| Short=LAAO ; Short=LAO ; AltName: Full=Interleukin-4-induced protein 1 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN115044567A (zh) | 2022-09-13 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Taguchi et al. | A search for novel cancer/testis antigens in lung cancer identifies VCX/Y genes, expanding the repertoire of potential immunotherapeutic targets | |
| CN107287174B (zh) | 肝癌标志物oxct1及其在肝癌诊断、治疗以及预后中的应用 | |
| CN111534592A (zh) | 一种研究m6A阅读器YTHDF1在卵巢癌发生中的功能和机制的方法 | |
| Chen et al. | CDK2 inhibition enhances antitumor immunity by increasing IFN response to endogenous retroviruses | |
| CN113908283A (zh) | Prmt5抑制剂及其与pd-l1抗体阻断剂联合在治疗肺癌上的应用 | |
| CN115044567B (zh) | 一种肺癌的代谢免疫靶点il4i1及其应用 | |
| CN113230404B (zh) | Sage1抑制剂在制备药物或试剂盒中的用途 | |
| JP2023505726A (ja) | 配列決定のための核酸を得る方法 | |
| CN110237257B (zh) | Ube3a泛素化PP2A激活因子PTPA在治疗天使综合症和孤独症中的应用 | |
| CN113876953A (zh) | 新型细胞衰老干预靶点及其在化疗抗癌中的靶向应用 | |
| CN111518899A (zh) | Nudt21基因在制备治疗肺癌药物中的应用 | |
| CN115837079A (zh) | Igf2bp1高表达在食管癌检测和治疗中的应用 | |
| CN112143805B (zh) | Rit1在肝细胞癌的诊断和治疗中的应用 | |
| CN116472349A (zh) | Line1的抑制剂及其用途 | |
| CN114099681A (zh) | Akt/stat3作为免疫检查点抑制剂的靶点的用途 | |
| CN114989286B (zh) | Ddup作为肿瘤耐药检测、治疗及预后分子靶点的应用 | |
| US10865415B2 (en) | Prevention, diagnosis and treatment of cancer overexpressing GPR160 | |
| CN112656805A (zh) | 抑制ythdf1活性的物质在制备预防或治疗胃癌产品中的应用 | |
| Zheng et al. | Characterizing the role of Tupaia DNA damage inducible transcript 3 (DDIT3) gene in viral infections | |
| CN111751532B (zh) | Piezo1蛋白作为食管癌标志物的应用 | |
| CN114058699B (zh) | Ppdpf在胰腺癌诊断及药物制备中的应用 | |
| CN114949217B (zh) | 癌症靶标及其应用 | |
| CN115814089B (zh) | Mt1g蛋白抑制剂作为靶点在制备治疗肾癌药物中的应用 | |
| CN116942796B (zh) | 一种长链非编码rna及其编码多肽或其检测试剂在制备诊断和/或治疗肾透明细胞癌产品中的应用 | |
| CN114949218B (zh) | 一种pd-l1调控剂及其应用 |
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 | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |