CN115778984A - Method and application for inducing transplantation immune tolerance and transplantation tolerance animal model - Google Patents
Method and application for inducing transplantation immune tolerance and transplantation tolerance animal model Download PDFInfo
- Publication number
- CN115778984A CN115778984A CN202310134489.0A CN202310134489A CN115778984A CN 115778984 A CN115778984 A CN 115778984A CN 202310134489 A CN202310134489 A CN 202310134489A CN 115778984 A CN115778984 A CN 115778984A
- Authority
- CN
- China
- Prior art keywords
- transplantation
- day
- rapa
- tolerance
- toldc
- 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
- 238000002054 transplantation Methods 0.000 title claims abstract description 72
- 238000000034 method Methods 0.000 title claims abstract description 40
- 230000006058 immune tolerance Effects 0.000 title claims abstract description 38
- 230000001939 inductive effect Effects 0.000 title claims abstract description 19
- 238000010171 animal model Methods 0.000 title claims abstract description 16
- 210000004443 dendritic cell Anatomy 0.000 claims abstract description 20
- 230000000638 stimulation Effects 0.000 claims abstract description 17
- 230000003614 tolerogenic effect Effects 0.000 claims abstract description 12
- 241000124008 Mammalia Species 0.000 claims abstract description 10
- 230000002980 postoperative effect Effects 0.000 claims abstract description 10
- 210000004185 liver Anatomy 0.000 claims description 32
- 210000003289 regulatory T cell Anatomy 0.000 claims description 29
- 210000004027 cell Anatomy 0.000 claims description 26
- 238000002360 preparation method Methods 0.000 claims description 11
- 108010017213 Granulocyte-Macrophage Colony-Stimulating Factor Proteins 0.000 claims description 10
- 238000001356 surgical procedure Methods 0.000 claims description 10
- 210000000056 organ Anatomy 0.000 claims description 9
- 108090000978 Interleukin-4 Proteins 0.000 claims description 8
- ZAHRKKWIAAJSAO-UHFFFAOYSA-N rapamycin Natural products COCC(O)C(=C/C(C)C(=O)CC(OC(=O)C1CCCCN1C(=O)C(=O)C2(O)OC(CC(OC)C(=CC=CC=CC(C)CC(C)C(=O)C)C)CCC2C)C(C)CC3CCC(O)C(C3)OC)C ZAHRKKWIAAJSAO-UHFFFAOYSA-N 0.000 claims description 7
- 229960002930 sirolimus Drugs 0.000 claims description 7
- QFJCIRLUMZQUOT-HPLJOQBZSA-N sirolimus Chemical compound C1C[C@@H](O)[C@H](OC)C[C@@H]1C[C@@H](C)[C@H]1OC(=O)[C@@H]2CCCCN2C(=O)C(=O)[C@](O)(O2)[C@H](C)CC[C@H]2C[C@H](OC)/C(C)=C/C=C/C=C/[C@@H](C)C[C@@H](C)C(=O)[C@H](OC)[C@H](O)/C(C)=C/[C@@H](C)C(=O)C1 QFJCIRLUMZQUOT-HPLJOQBZSA-N 0.000 claims description 7
- 210000001519 tissue Anatomy 0.000 claims description 7
- 230000008859 change Effects 0.000 claims description 6
- 241000282414 Homo sapiens Species 0.000 claims description 5
- 238000012258 culturing Methods 0.000 claims description 4
- 230000001105 regulatory effect Effects 0.000 claims description 4
- 230000001502 supplementing effect Effects 0.000 claims description 4
- 239000001963 growth medium Substances 0.000 claims description 3
- 210000002216 heart Anatomy 0.000 claims description 3
- 210000003734 kidney Anatomy 0.000 claims description 3
- 210000005259 peripheral blood Anatomy 0.000 claims description 3
- 239000011886 peripheral blood Substances 0.000 claims description 3
- 241000282693 Cercopithecidae Species 0.000 claims description 2
- 241000283073 Equus caballus Species 0.000 claims description 2
- 241000282326 Felis catus Species 0.000 claims description 2
- 241000283973 Oryctolagus cuniculus Species 0.000 claims description 2
- 241000282577 Pan troglodytes Species 0.000 claims description 2
- 241001494479 Pecora Species 0.000 claims description 2
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N Phenol Chemical compound OC1=CC=CC=C1 ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 claims description 2
- 238000004115 adherent culture Methods 0.000 claims description 2
- 210000001185 bone marrow Anatomy 0.000 claims description 2
- 239000012636 effector Substances 0.000 claims description 2
- 210000004153 islets of langerhan Anatomy 0.000 claims description 2
- 239000007788 liquid Substances 0.000 claims description 2
- 210000004072 lung Anatomy 0.000 claims description 2
- 210000002901 mesenchymal stem cell Anatomy 0.000 claims description 2
- 210000000496 pancreas Anatomy 0.000 claims description 2
- 239000002243 precursor Substances 0.000 claims description 2
- 210000000813 small intestine Anatomy 0.000 claims description 2
- 238000011476 stem cell transplantation Methods 0.000 claims description 2
- 102000004457 Granulocyte-Macrophage Colony-Stimulating Factor Human genes 0.000 claims 5
- 102100034922 T-cell surface glycoprotein CD8 alpha chain Human genes 0.000 description 44
- 241000700159 Rattus Species 0.000 description 35
- 238000001802 infusion Methods 0.000 description 26
- 230000004083 survival effect Effects 0.000 description 20
- 230000000694 effects Effects 0.000 description 12
- 230000006698 induction Effects 0.000 description 10
- 230000001154 acute effect Effects 0.000 description 8
- 230000005764 inhibitory process Effects 0.000 description 8
- 230000008901 benefit Effects 0.000 description 7
- 230000007774 longterm Effects 0.000 description 7
- 238000011160 research Methods 0.000 description 7
- 210000002865 immune cell Anatomy 0.000 description 6
- 239000003018 immunosuppressive agent Substances 0.000 description 6
- 230000002401 inhibitory effect Effects 0.000 description 6
- 238000011282 treatment Methods 0.000 description 6
- 102100039620 Granulocyte-macrophage colony-stimulating factor Human genes 0.000 description 5
- 241000699666 Mus <mouse, genus> Species 0.000 description 5
- 241001506137 Rapa Species 0.000 description 5
- 230000034994 death Effects 0.000 description 5
- 238000000684 flow cytometry Methods 0.000 description 5
- 238000002156 mixing Methods 0.000 description 5
- 230000028327 secretion Effects 0.000 description 5
- 230000002269 spontaneous effect Effects 0.000 description 5
- 102000006354 HLA-DR Antigens Human genes 0.000 description 4
- 108010058597 HLA-DR Antigens Proteins 0.000 description 4
- 238000006243 chemical reaction Methods 0.000 description 4
- 238000011161 development Methods 0.000 description 4
- 238000002474 experimental method Methods 0.000 description 4
- 238000007490 hematoxylin and eosin (H&E) staining Methods 0.000 description 4
- 230000001506 immunosuppresive effect Effects 0.000 description 4
- 229940125721 immunosuppressive agent Drugs 0.000 description 4
- 239000000243 solution Substances 0.000 description 4
- 206010062016 Immunosuppression Diseases 0.000 description 3
- 102000013530 TOR Serine-Threonine Kinases Human genes 0.000 description 3
- 108010065917 TOR Serine-Threonine Kinases Proteins 0.000 description 3
- 238000004458 analytical method Methods 0.000 description 3
- 239000000427 antigen Substances 0.000 description 3
- 108091007433 antigens Proteins 0.000 description 3
- 102000036639 antigens Human genes 0.000 description 3
- 238000001514 detection method Methods 0.000 description 3
- 239000005556 hormone Substances 0.000 description 3
- 229940088597 hormone Drugs 0.000 description 3
- 210000004698 lymphocyte Anatomy 0.000 description 3
- 238000004519 manufacturing process Methods 0.000 description 3
- 239000012528 membrane Substances 0.000 description 3
- 230000002035 prolonged effect Effects 0.000 description 3
- 102000005962 receptors Human genes 0.000 description 3
- 238000010186 staining Methods 0.000 description 3
- 239000006228 supernatant Substances 0.000 description 3
- 206010061218 Inflammation Diseases 0.000 description 2
- 241001465754 Metazoa Species 0.000 description 2
- 241000282898 Sus scrofa Species 0.000 description 2
- 239000012620 biological material Substances 0.000 description 2
- 238000007664 blowing Methods 0.000 description 2
- 230000004069 differentiation Effects 0.000 description 2
- 230000003828 downregulation Effects 0.000 description 2
- GNBHRKFJIUUOQI-UHFFFAOYSA-N fluorescein Chemical compound O1C(=O)C2=CC=CC=C2C21C1=CC=C(O)C=C1OC1=CC(O)=CC=C21 GNBHRKFJIUUOQI-UHFFFAOYSA-N 0.000 description 2
- 230000006870 function Effects 0.000 description 2
- 229960003444 immunosuppressant agent Drugs 0.000 description 2
- 230000001861 immunosuppressant effect Effects 0.000 description 2
- 230000008595 infiltration Effects 0.000 description 2
- 238000001764 infiltration Methods 0.000 description 2
- 230000004054 inflammatory process Effects 0.000 description 2
- 239000003550 marker Substances 0.000 description 2
- 230000007246 mechanism Effects 0.000 description 2
- 230000001575 pathological effect Effects 0.000 description 2
- 230000035755 proliferation Effects 0.000 description 2
- 230000009467 reduction Effects 0.000 description 2
- 230000003248 secreting effect Effects 0.000 description 2
- 239000000725 suspension Substances 0.000 description 2
- 238000002560 therapeutic procedure Methods 0.000 description 2
- 238000011269 treatment regimen Methods 0.000 description 2
- 102000010400 1-phosphatidylinositol-3-kinase activity proteins Human genes 0.000 description 1
- 241000283690 Bos taurus Species 0.000 description 1
- 102000004127 Cytokines Human genes 0.000 description 1
- 108090000695 Cytokines Proteins 0.000 description 1
- 206010016654 Fibrosis Diseases 0.000 description 1
- 229920001917 Ficoll Polymers 0.000 description 1
- 108010043121 Green Fluorescent Proteins Proteins 0.000 description 1
- 102000018713 Histocompatibility Antigens Class II Human genes 0.000 description 1
- 108010027412 Histocompatibility Antigens Class II Proteins 0.000 description 1
- 101000738771 Homo sapiens Receptor-type tyrosine-protein phosphatase C Proteins 0.000 description 1
- 101000914484 Homo sapiens T-lymphocyte activation antigen CD80 Proteins 0.000 description 1
- 102100040061 Indoleamine 2,3-dioxygenase 1 Human genes 0.000 description 1
- 102100022297 Integrin alpha-X Human genes 0.000 description 1
- 229930191564 Monensin Natural products 0.000 description 1
- GAOZTHIDHYLHMS-UHFFFAOYSA-N Monensin A Natural products O1C(CC)(C2C(CC(O2)C2C(CC(C)C(O)(CO)O2)C)C)CCC1C(O1)(C)CCC21CC(O)C(C)C(C(C)C(OC)C(C)C(O)=O)O2 GAOZTHIDHYLHMS-UHFFFAOYSA-N 0.000 description 1
- 208000001388 Opportunistic Infections Diseases 0.000 description 1
- 108091007960 PI3Ks Proteins 0.000 description 1
- 241000009328 Perro Species 0.000 description 1
- 102100037422 Receptor-type tyrosine-protein phosphatase C Human genes 0.000 description 1
- 101150052966 SIGLEC11 gene Proteins 0.000 description 1
- 102100032855 Sialoadhesin Human genes 0.000 description 1
- 230000006044 T cell activation Effects 0.000 description 1
- 230000033540 T cell apoptotic process Effects 0.000 description 1
- 230000006052 T cell proliferation Effects 0.000 description 1
- 102100027222 T-lymphocyte activation antigen CD80 Human genes 0.000 description 1
- 206010060872 Transplant failure Diseases 0.000 description 1
- 206010052779 Transplant rejections Diseases 0.000 description 1
- QYSXJUFSXHHAJI-XFEUOLMDSA-N Vitamin D3 Natural products C1(/[C@@H]2CC[C@@H]([C@]2(CCC1)C)[C@H](C)CCCC(C)C)=C/C=C1\C[C@@H](O)CCC1=C QYSXJUFSXHHAJI-XFEUOLMDSA-N 0.000 description 1
- 208000027418 Wounds and injury Diseases 0.000 description 1
- 239000003124 biologic agent Substances 0.000 description 1
- 230000029918 bioluminescence Effects 0.000 description 1
- 238000005415 bioluminescence Methods 0.000 description 1
- 230000000903 blocking effect Effects 0.000 description 1
- 230000037396 body weight Effects 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 201000011510 cancer Diseases 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 239000006285 cell suspension Substances 0.000 description 1
- 238000002659 cell therapy Methods 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 230000001684 chronic effect Effects 0.000 description 1
- 230000007882 cirrhosis Effects 0.000 description 1
- 208000019425 cirrhosis of liver Diseases 0.000 description 1
- 238000011281 clinical therapy Methods 0.000 description 1
- 238000010835 comparative analysis Methods 0.000 description 1
- 238000012790 confirmation Methods 0.000 description 1
- 238000010226 confocal imaging Methods 0.000 description 1
- 230000001276 controlling effect Effects 0.000 description 1
- 210000004087 cornea Anatomy 0.000 description 1
- 238000005520 cutting process Methods 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 230000002950 deficient Effects 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 210000003162 effector t lymphocyte Anatomy 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000029142 excretion Effects 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 238000005206 flow analysis Methods 0.000 description 1
- 238000000227 grinding Methods 0.000 description 1
- 238000003384 imaging method Methods 0.000 description 1
- 230000001900 immune effect Effects 0.000 description 1
- 230000003832 immune regulation Effects 0.000 description 1
- 230000008629 immune suppression Effects 0.000 description 1
- 238000009169 immunotherapy Methods 0.000 description 1
- 230000001976 improved effect Effects 0.000 description 1
- 208000014674 injury Diseases 0.000 description 1
- PGHMRUGBZOYCAA-UHFFFAOYSA-N ionomycin Natural products O1C(CC(O)C(C)C(O)C(C)C=CCC(C)CC(C)C(O)=CC(=O)C(C)CC(C)CC(CCC(O)=O)C)CCC1(C)C1OC(C)(C(C)O)CC1 PGHMRUGBZOYCAA-UHFFFAOYSA-N 0.000 description 1
- PGHMRUGBZOYCAA-ADZNBVRBSA-N ionomycin Chemical compound O1[C@H](C[C@H](O)[C@H](C)[C@H](O)[C@H](C)/C=C/C[C@@H](C)C[C@@H](C)C(/O)=C/C(=O)[C@@H](C)C[C@@H](C)C[C@@H](CCC(O)=O)C)CC[C@@]1(C)[C@@H]1O[C@](C)([C@@H](C)O)CC1 PGHMRUGBZOYCAA-ADZNBVRBSA-N 0.000 description 1
- 238000002372 labelling Methods 0.000 description 1
- 230000003908 liver function Effects 0.000 description 1
- 210000005228 liver tissue Anatomy 0.000 description 1
- 238000012423 maintenance Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 208000030159 metabolic disease Diseases 0.000 description 1
- 238000000386 microscopy Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 229960005358 monensin Drugs 0.000 description 1
- GAOZTHIDHYLHMS-KEOBGNEYSA-N monensin A Chemical compound C([C@@](O1)(C)[C@H]2CC[C@@](O2)(CC)[C@H]2[C@H](C[C@@H](O2)[C@@H]2[C@H](C[C@@H](C)[C@](O)(CO)O2)C)C)C[C@@]21C[C@H](O)[C@@H](C)[C@@H]([C@@H](C)[C@@H](OC)[C@H](C)C(O)=O)O2 GAOZTHIDHYLHMS-KEOBGNEYSA-N 0.000 description 1
- 210000005087 mononuclear cell Anatomy 0.000 description 1
- 230000007170 pathology Effects 0.000 description 1
- 230000037361 pathway Effects 0.000 description 1
- 210000003819 peripheral blood mononuclear cell Anatomy 0.000 description 1
- 230000000144 pharmacologic effect Effects 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 101150063780 spp1 gene Proteins 0.000 description 1
- 230000004936 stimulating effect Effects 0.000 description 1
- 230000009885 systemic effect Effects 0.000 description 1
- 238000013518 transcription Methods 0.000 description 1
- 230000035897 transcription Effects 0.000 description 1
- 230000009466 transformation Effects 0.000 description 1
- QYSXJUFSXHHAJI-YRZJJWOYSA-N vitamin D3 Chemical compound C1(/[C@@H]2CC[C@@H]([C@]2(CCC1)C)[C@H](C)CCCC(C)C)=C\C=C1\C[C@@H](O)CCC1=C QYSXJUFSXHHAJI-YRZJJWOYSA-N 0.000 description 1
- 235000005282 vitamin D3 Nutrition 0.000 description 1
- 239000011647 vitamin D3 Substances 0.000 description 1
- 229940021056 vitamin d3 Drugs 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
Images
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
- A61K35/14—Blood; Artificial blood
- A61K35/15—Cells of the myeloid line, e.g. granulocytes, basophils, eosinophils, neutrophils, leucocytes, monocytes, macrophages or mast cells; Myeloid precursor cells; Antigen-presenting cells, e.g. dendritic cells
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P37/00—Drugs for immunological or allergic disorders
- A61P37/02—Immunomodulators
- A61P37/06—Immunosuppressants, e.g. drugs for graft rejection
-
- 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
- G01N33/569—Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses
- G01N33/56966—Animal cells
-
- 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
- G01N33/577—Immunoassay; Biospecific binding assay; Materials therefor involving monoclonal antibodies binding reaction mechanisms characterised by the use of monoclonal antibodies; monoclonal antibodies per se are classified with their corresponding antigens
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2469/00—Immunoassays for the detection of microorganisms
- G01N2469/10—Detection of antigens from microorganism in sample from host
Abstract
The invention provides a method for inducing transplantation immune tolerance, which is characterized in that the method comprises the step of administering mTOR-targeted tolerogenic dendritic cells (Rapa-tolDC) to transplanted mammals by adopting a scheme of preoperative pre-stimulation and postoperative feedback. Also provides application of the Rapa-tolDC in preparing a transplantation tolerance animal model and the transplantation tolerance animal model prepared by the method.
Description
Technical Field
The invention relates to a method for inducing transplantation immune tolerance, application and a transplantation tolerance animal model. In particular, the invention relates to a method for inducing transplantation immune tolerance by using mTOR-targeted tolerogenic dendritic cells (Rapa-tolDC), application of the Rapa-tolDC in preparation of a transplantation tolerance animal model and the transplantation tolerance animal model prepared by the method.
Background
Organ transplantation is the best treatment for end-stage organ disease, but graft rejection is the major cause of the impact on the quality of life and long-term survival of recipients. The continued development of immunosuppressive agents in the last 30 years has significantly improved the short-term survival benefit of transplant recipients, but the long-term survival rate has not changed significantly. The currently clinically used immunosuppressive schemes also have many defects, such as chronic graft failure caused by long-term immunosuppression, metabolic diseases, graft injury, opportunistic infection, severe threat to long-term survival of patients caused by malignant tumors, and the requirement of lifetime administration of immunosuppressive agents.
Induction of immune tolerance is the most ideal treatment option for controlling rejection and maintaining long-term survival of recipients without immune control. Successful establishment of immune tolerance can not only solve complications and side effects caused by long-term application of immunosuppressive agents, but also maintain long-term survival of transplants, and has significant medical and economic benefits. However, there is no operational transplantation immune tolerance scheme applicable to clinical application, and a novel technical scheme for safely and effectively inducing organ transplantation immune tolerance is developed, so that a receptor is promoted to establish an immune tolerance state specific to a donor, and the immune tolerance scheme is successfully applied to human beings in a transformation way, and the method has important research value, and huge clinical application prospect and social and economic benefits.
Regulatory dendritic cells (toldcs) have a negative immune regulation function, and play a role in inducing immune tolerance by inhibiting T cell proliferation, inhibiting antigen-specific T cell activation, mediating T cell apoptosis, inducing regulatory T cells, and the like. Induction of immune tolerance using adoptive reinfusion therapy of tolDC immune cells is the hot spot of current research. The small dose of immunosuppressant can improve the survival of transplanted liver, kidney and heart by combining or independently returning tolDC; the PBMC derived tolDC IS safely and effectively induced by the IL-10 and the vitamin D3 combined with the cytokines through single feedback of a living liver transplantation patient, but IS withdrawal IS not solved, and the end of immune tolerance IS reached. Reinfusion of autologous or syngeneic toldcs, whether or not loaded with donor antigens, can improve graft survival. These all suggest that adoptive infusion of toldcs is expected to be an ideal solution for immune tolerance. However, there are few reports related to toldcs that can be used for clinical therapy, have strong inhibition and are functionally stable.
Induction of toldcs with donor-specific immunosuppression is a primary problem facing current clinical studies. Among various currently and clinically used immunosuppressive agents, rapa can induce anti-mature tolDC with tolerance, promote differentiation, proliferation and suppressivity of Tregs, has no effect on CsA, and has inhibitory effect on hormone and MPA; rapamycin (Rapa) withdrawal has the advantage of promoting immune tolerance in long-lived liver transplants, although the sample size is small, suggesting that Rapa may have a unique advantage in inducing immune tolerance; rapa can induce the anti-mature tolDC with stable function and promote the differentiation of Treg, while the CsA which is commonly used in clinic has no effect, and the hormone and MPA have inhibitory effect. However, no report is available on the clinical research of the Rapa-induced tolDC, and the research on animal liver transplantation is less.
Patent CN202210978763.8, publication CN115044553A, discloses mTOR-targeted tolerocyte dendritic cells, and a preparation method and application thereof, and tolDC with stable tolerance is obtained.
However, how to determine the dose, time and frequency of cell therapy to establish a standardized treatment regimen remains an important issue facing tolDC immunotherapy.
Disclosure of Invention
It is an object of the present invention to provide a method for inducing transplantation immune tolerance.
The invention also aims to provide application of the Rapa-tolDC in preparation of a transplantation tolerance animal model.
It is still another object of the present invention to provide an animal model of graft tolerance prepared by the above method.
On the basis of the previous application CN202210978763.8 of the same applicant, the invention further researches the effects of Rapa-tolDC adoptive feedback reversal or rejection inhibition and immune tolerance induction, and simultaneously adopts a control tolDC (prior scheme) for comparison to determine the treatment hopefully inducing immune tolerance successfullyThe scheme can greatly reduce the death rate and obviously prolong the survival time of the subject. Further, the mechanism of immune tolerance was investigated, and it was found that MHC-II was highly expressed + CD8 of (1) + CD45RC low/- Tregs (MHC-II + CD8+ Tregs) are the major and critical cell subset for donor-specific immunosuppression, and furthermore, MHC-II was found to be derived from Rapa-tolDC, rather than CD8+ CD45 RClow/-Tregs expressed by themselves, with donor-specific suppressive effects. Thus forming the present invention.
In particular, in one aspect, the invention provides a method of inducing transplantation immune tolerance comprising administering mTOR-targeted tolerant dendritic cells (Rapa-toldcs) to a transplanted mammal using a pre-operative pre-stimulation and post-operative reinfusion protocol.
The mTOR-targeted tolerant dendritic cell (Rapa-tolDC) is prepared by the method described in patent CN202210978763.8, has a transcription profile of Siglec1 and Spp1 gene down regulation, is deficient in PI3K/mTOR expression, and has a selective negative regulation effect. In addition, the Rapa-tolDC expresses a DC marker CD11c, expresses low surface co-stimulatory molecules CD80, CD86 and MHC-II, and secretes relatively high IL-10 and low INF-gamma compared with mature DC; can induce the generation of various types of regulatory immune cells, and can generate CD8 with donor specificity + Regulatory T cells.
According to a particular embodiment of the invention, the method is performed using a pre-operative pre-stimulation plus 1 to 3 reinfusion cycles within 1 month after surgery.
According to a particular embodiment of the invention, the method is carried out using a pre-stimulation 7 days before surgery + reinfusion the day after surgery.
According to a particular embodiment of the invention, the method is performed using a pre-stimulation 7 days before operation plus a reinfusion 7 days after operation.
According to a particular embodiment of the invention, the method is performed using a pre-stimulation schedule of 7 days before surgery plus a reinfusion schedule of 7, 14 and 28 days after surgery.
According to a particular embodiment of the invention, when said mammal is a rat, according to DadaThe effective cell number per time of reinfusion is 1X 10 according to the body weight of the mouse and the actual condition of the operation 6 -1×10 7 . When the mammal is a mammal other than a rat, the effective cell number per reinfusion can be appropriately converted by those skilled in the art, for example, referring to the pharmacological experimental methodology (2002) compiled by the professor Xu Shuyun, the equivalent dose of the rat is about 6.3 times that of the human in terms of a unit weight dosimeter calculated according to the equivalent dose coefficient reduction algorithm.
According to a specific embodiment of the present invention, the Rapa-tolDC induces high expression of MHC-II in the graft + CD8 of (1) + CD45RC low/- Production of Tregs (MHC-II + CD8+ Tregs).
According to a particular embodiment of the invention, the transplantation comprises cell transplantation, organ transplantation or tissue transplantation; wherein the cell transplantation comprises stem cell transplantation, regulatory cell transplantation, islet cell transplantation or effector cell transplantation; organ transplantation includes kidney transplantation, liver transplantation, heart transplantation, small intestine transplantation, lung transplantation, pancreas transplantation, or combined organ transplantation; the tissue transplantation includes cornea transplantation, limb transplantation or face transplantation, and preferably liver transplantation.
According to a particular embodiment of the invention, the mammal is a rat, mouse, pig, rabbit, dog, cat, horse, cow, sheep, monkey, chimpanzee or human.
According to a specific embodiment of the present invention, the Rapa-tolDC is prepared by a method comprising:
taking bone marrow mesenchymal stem cells or peripheral blood PMBC, carrying out adherent culture by using a phenol red-free culture solution containing 1% -10% FBS to obtain precursor cells of the DC, and adding a complete culture medium containing GM-CSF and IL-4; culturing for full change on day 2, half change on day 4, supplementing cell factors GM-CSF and IL-4, maintaining concentration, and adding rapamycin when changing liquid on day 2 and day 4; the culture was continued by day 6, half-way changing of the medium, supplementing GM-CSF to maintain the concentration unchanged, and adding LPS.
According to a preferred embodiment of the invention, the concentration of GM-CSF is between 1 and 40ng/ml, the concentration of IL-4 is between 0.5 and 20ng/ml, the concentration of rapamycin is between 1 and 40nM, the concentration of LPS is between 0.01 and 1. Mu.g/ml,
according to a preferred embodiment of the invention, the ratio of the concentration of GM-CSF to IL-4 is 2:1; rapamycin was added at 10nM concentrations at day 2 and day 4 changes, respectively, and LPS was added at 0.05. Mu.g/mL at day 6.
On the other hand, the invention also provides application of the Rapa-tolDC defined by the method in preparation of a transplantation tolerance animal model.
In yet another aspect, the present invention also provides an animal model of graft tolerance prepared by the above method.
In yet another aspect, the present invention also provides a method for preparing an animal model for transplant tolerance by the above method. For example, it can be prepared by performing the method of the present invention (i.e., pre-stimulation and post-operative reinfusion) by administering mTOR-targeted tolerogenic dendritic cells (Rapa-toldcs) to an existing animal model of acute rejection.
In the invention, the effects of Rapa-tolDC adoptive reversal or rejection inhibition and immune tolerance induction are further researched, and meanwhile, a control tolDC (the conventional scheme) is adopted for comparison, so that the most hopeful treatment scheme for successfully inducing immune tolerance is determined, the death rate can be greatly reduced, and the survival time of a subject is obviously prolonged. Meanwhile, the method can be used for simply and quickly preparing a transplantation tolerance animal model and providing an important model animal for the mechanism research of tolDC treatment.
Drawings
The accompanying drawings, which are incorporated in and constitute a part of this specification, are included to provide a further understanding of the invention and to enable other features, objects and advantages of the invention to be more fully apparent. The drawings and their description illustrate exemplary embodiments of the invention and do not limit the scope of the invention unduly.
FIG. 1 is a graph of the results of single infusion Rapa-tolDC rat transplanted liver HE staining compared to control groups.
Figure 2 is a graph of rat survival time for single infusions of different resistant DCs.
Fig. 3 is a schematic flow chart of the technical scheme of inducing the immunological tolerance of the liver transplantation rat by the adoptive infusion of the Rapa-tolDC according to the embodiment of the invention.
FIG. 4 is a graph of survival time for Rapa-tolDC adoptively infused rats according to the technical scheme of FIG. 3.
FIG. 5 is a graph showing the results of HE staining of transplanted liver in rats adoptively infused with Rapa-tolDC in the example of FIG. 4.
FIG. 6 is a graph showing the result of adoptive infusion of Rapa-tolDC rats transplanted with liver MHC-II + CD8+ Treg and their secretion of IL-10 and INF-gamma in the example of FIG. 5.
FIG. 7 is a graph of the results of confocal imaging of MHC-II + CD8+ Treg surface MHC-II molecule sources.
FIG. 8 is a graph of the results of flow analysis of the induction of MHC-II + CD8+ Treg production by Rapa-tolDC.
FIG. 9 is a graph of flow-through results of the donor-specific inhibition of MHC-II + CD8+ Tregs induced in the examples.
FIG. 10 is a graph showing the results of HLA-DR + CD8+ Treg expression in spontaneously clinically tolerated and stably surviving liver transplant patients.
Detailed Description
In order to clearly understand the technical features, objects and advantages of the present invention, the technical solutions of the present invention will be described in more detail with reference to the specific embodiments and the accompanying drawings. It is obvious that the described embodiments are only examples of a part of the present invention, and the present invention is not limited thereto. All other embodiments, which can be obtained by a person skilled in the art without making creative efforts based on the embodiments of the present invention, shall fall within the protection scope of the present invention.
It should be noted that the obtaining route of various biological reagents and materials used in the examples is only to provide a route of experimental acquisition to achieve the purpose of specific disclosure of the invention, and should not be a limitation to the source of the biological materials of the invention. In fact, any biological agent or material that can be obtained without law or ethical reasons, including isolated cells from a mammal or human such as rat, mouse, pig, or the like, or obtained from a cell bank, or purchased commercially, or prepared according to available literature, can be used instead as suggested in the examples of the present invention.
The method operations not specifically mentioned in the examples were carried out according to the conventional operations of the prior art or the operations suggested by the manufacturer's specifications.
Example 1 preparation of tolerogenic immune cell preparation
Preparation method mTOR-targeting tolerogenic dendritic cells (Rapa-toldcs), which are immune cell preparations with tolerogenic properties, were prepared exactly as described in CN202210978763.8, publication CN115044553a, example 1.
Example 2 analysis of Effect of different tolerogenic immune cell preparations on Reversal of rejection or Induction of tolerance
In this example, the effects of adoptive reinfusion reversal or rejection inhibition and immune tolerance induction of the tolerogenic immune cell preparation (Rapa-tolDC) of the present invention were examined, and compared with a control tolDC (conventional protocol).
The first scheme is as follows: single infusion of mTOR-targeted tolerant DC (dendritic cell) to relieve rejection degree of transplanted liver and prolong survival time
Using the targeted mTOR prepared in example 1, the effective cell number per reinfusion was 1X 10 according to the pre-stimulation on day 7 before surgery and reinfusion on day after surgery, based on the weight of the rat and the actual condition of the surgery 6 -1×10 7 The median survival time of transplanted liver rats was extended to 37 days, while the median survival time of transplanted liver rats was extended to 32 days after tolDC infusion in the control group. HE staining suggested a significant reduction in lymphocyte infiltration in the transplanted livers compared to the acute-row transplanted rats (a-C in fig. 1). The above suggests that adoptive infusion of the liver-transplanted rat with the property-tolerant DC can inhibit acute rejection reaction safely and practically.
Scheme two is as follows: changes the time point of single infusion, effectively prolongs the survival time of rats and reduces the death rate in the perioperative period
Using the targeted mTOR prepared in example 1, the effective number of cells per reinfusion was 1X 10 according to the pre-operative 7 days pre-stimulation plus postoperative 7 days reinfusion protocol, based on the weight of the rat and the actual conditions of the operation 6 -1×10 7 In the operation, the median survival time of transplanted liver rats is prolonged to 45 days by adopting small dose hormone intervention, no mortality rate occurs in the perioperative period (figure 2, including perioperative death), the death caused by systemic immune inflammatory reaction induced by adoptive infusion in the day of the operation is avoided (the mortality rate is 10-15% in the day of infusion in a scheme I and a control group), and the pathological degree of acute rejection reaction is relatively light when HE staining is carried out (D in figure 1).
These all suggest that further confirmation that mTOR-targeted tolerant DC single reinfusion is safe and feasible, and changing the time point of infusion can effectively improve the safety of infusion.
The third scheme is as follows: the scheme of pre-operation pre-stimulation and three times of postoperative adoptive infusions has the potential of inducing immune tolerance
In order to further verify the effectiveness of Rapa-tolDC in inducing immune tolerance and search for a feasible scheme for establishing immune tolerance by adoptive infusion of mTOR-targeted tolDC, the safety and effectiveness of different adoptive infusion treatment schemes are explored on the basis of the single-adoptive-infusion Rapa-tolDC research. BN is used as a receptor, lewis is used as a donor to establish a rat acute excretion model, lewis is used as a receptor, BN is used as a donor to establish a spontaneous rat liver transplantation model, and Rapa-tolDC is used as an intervention means. We change the time point of the return infusion into 7 days after the operation, 14 days and 28 days, the scheme in the operation is not changed (figure 3), and it can be seen that compared with the single-time after the operation, the survival time of the Rapa-tolDC adoptive infusion rat is obviously prolonged, the median survival time can reach 65 days (9 days in the middle of the emergent rats) (figure 4), the rat transplantation liver of the Rapa-tolDC adoptive infusion rat is visible to naked eyes and is close to normal to the transplantation liver of the spontaneous tolerant rat, and there is no rejection reaction expression visible to naked eyes; pathological staining of transplanted liver tissue shows that the lobular structure of liver is basically normal, the degree of inflammatory reaction is light, the infiltration of lymphocyte is less, the liver lobular structure is mainly concentrated in the region of the sink (figure 5), the acute rejection reaction performance confirmed by pathology is not existed under the microscope, and the RAI index is not different from that of the spontaneous immune tolerance group (1.75 +/-0.957)vs.1.25±0.50）。
All of the above suggests that the present invention based on the adoptive infusion therapy of mTOR-targeted tolerant DCs can induce the development of immune tolerance.
Example 3 infusion of different tolerogenic dendritic cell rats high expression of CD8 in transplanted liver + CD45RC low/- Tregs are the basis for the development of immune tolerance
Cutting fresh rat transplantation tissue, grinding, diluting tissue cell culture solution RMPI-1640 to obtain suspension, filtering with 200 mesh screen for 2-3 times, extracting cell suspension, extracting mononuclear cells with tissue lymphocyte extract Ficoll centrifugation, washing with 1 XPBS once, discarding supernatant, adding 200 μ l 1 XPBS, mixing well, adding 5 μ l mouse rat-resistant monoclonal antibody FITC-CD8, PE-CD45RC and Percp-TCR alpha beta (purchased from BD company of America), mixing well, incubating at room temperature and dark for 15min, discarding supernatant, blowing and mixing well with 1 XPBS 500 μ l, detecting CD8 with flow cytometer to obtain suspension + CD45RC low/- Expression level of tregs.
Flow cytometry results showed that in case one, mTOR-targeted, tolerogenic DC rats were back-transfused with CD8 in transplanted liver as compared to control (tolDC) + CD45RC low/- Treg expression grade (92.8 +/-7.89)vs.89.6 +/-3.29); scheme two and scheme three transplantation of CD8 in liver + CD45RC low/- Treg expression was maintained between 92% and 95% on average. The detection result shows that the mTOR-targeted tolerogenic DC promotes the generation of immune tolerance and needs to maintain higher level of CD8 + Treg levels.
As indicated above, regimen three, "pre-operative pre-stimulation + three post-operative adoptive infusions" is the most promising treatment regimen for successfully inducing immune tolerance. In the following examples, comparative analysis was carried out on only protocol three and a control group, if not specified.
Example 4 high expression of MHC-II + CD8 of (1) + CD45RC low/- Tregs may be in the induction of liver transplantation immune tolerance in the main and key subset
+ + Adoptive infusion of Rapa-tolDC and high expression of IL-10 secreting MHC-IICD8 in spontaneously tolerated rat transplanted liver
low/- CD45RCTreg
The experimental procedure of example 3 was followed, before stainingFirstly stimulating for 4-6h by using monensin and ionomycin, performing membrane antibody staining scheme as before, breaking membrane, adding 5 μ l of mouse anti-rat monoclonal antibody AF647-IL-10 (purchased from BD company in America), mixing well, incubating at room temperature in dark for 60min, adding 2ml of membrane breaking buffer solution, centrifuging, discarding supernatant, blowing and mixing well with 1 × PBS 500 μ l, detecting MHC-II by flow cytometry + CD8 + CD45RC low/- Expression level of tregs.
The detection result shows that the high expression of MHC-II in the transplanted liver of the rat with adoptive infusion of Rapa-tolDC and spontaneous tolerance + CD8 + CD45RC low/- Tregs, which are hardly expressed in transplanted liver in acute rats; MHC-II in transplanted livers from control rats + CD8 + CD45RC low/- The level of tregs was also significantly reduced (a of fig. 6). Further analysis of IL-10 secretion level and INF-gamma ability revealed that the adoptive infusion of Rapa-tolDC and spontaneously tolerant rats expressed higher levels of MHC-II of IL-10 + CD8 + CD45RC low/- Treg, significantly higher than the acute rejection and control (B of fig. 6), while INF- γ levels were significantly reduced (C of fig. 6).
+ + low/- + low/- MHC-IICD8CD45RCTreg is the main source of IL-10 secretion in CD8CD45RCTreg, and - non-MHC-II subpopulations, a critical cell subpopulation for induction of tolerance。
In the invention patent application CN202210978763.8, we used mTOR-targeted tolerant DCs to induce the production of highly IL-10-secreting CD8 + CD45RC low/- Tregs, based on the results described above, were classified into positive and negative populations using MHC-II and, following flow-based detection using the same staining protocol as described above, analysis revealed that they were found to be CD 8-independent + CD45RC low Treg is also MHC-II + CD8 + Tregs, which secrete IL-10 with a mean level of over 60%, were further analyzed to find, although MHC-II + CD8 + The proportion of Tregs is only between 20% and 30%, but MHC-II + CD8 + The level of IL-10 secretion by Tregs is CD8 + CD45RC low Total level of IL-10 secretion in Tregs79.97% of (1), is MHC-II - CD8 + Tregs secrete more than 3.5 times the level of IL-10.
These suggest that MHC-II + CD8 + CD45RC low/- Tregs are a critical subset of cells that play an important role in the development of inducing immune tolerance. In the invention patent application CN202210978763.8, we found CD8 + CD45RC low/- Tregs specifically inhibit the proliferation of effector T cells, and based on these results, we believe that MHC-II expression is observed + CD8 of (1) + CD45RC low/- Treg is also CD8 + CD45RC low/- Tregs play a major subset of donor-specific immune suppression.
Example 5 MHC-II was derived from Rapa-tolDC, not CD8 + CD45RC low/- Tregs expressed by themselves, with donor-specific inhibitory action
We used PKH26 and CFSE bioluminescence marker to label Rapa-tolDC and CD8 respectively + CD45RC low/- Treg, after mixed culture for 7 days, observing the cells fused with each other under a laser confocal microscope; flow cytometry was used to detect the proportion of double-positive cell subsets labeled with bifluorescin, which expressed the proportion of MHC-II double-positive cell subsets.
CFSE labeled green fluorescent CD8 was observed by confocal laser microscopy imaging + CD45RC low The red Rapa-tolDC part marked by partial PKH26 is obtained on the surface of Treg, and the flow cytometry technology initially proves that after two kinds of cells marked by biological fluorescein are mixed, MHC-II + CD8 + The proportion of MHC-II molecules on the surface of Tregs derived from dendritic cells (FIG. 7) was significantly increased, 1-MT intervention inhibited expression, and 1-MT upregulated expression, all suggesting that surface MHC-II was derived from Rapa-tolDC (A in FIG. 8).
To avoid the effect of biofluorescin on the expression level, we repeated the above experiment without fluorescein labeling and found that although CD8 was present after blocking the IDO pathway using 1-MT + CD45RC low There was no significant difference in Treg expression, but MHC-II + CD8 + The level of tregs is significantly reduced; while the analogue of IDO 3-4,DAA intervenesCan then up-regulate CD8 + CD45RC low Treg、MHC-II + CD8 + The level of Tregs (B-D of FIG. 8).
According to the experimental procedure in the invention patent application CN202210978763.8, donor specific inhibition experiments were repeated, and it was found that MHC-II + CD8+ Treg has strong donor specific inhibition effect, and the specific inhibition effect was broken by third party derived antigen stimulation (fig. 9). All the above suggest that MHC-II is derived from Rapa-tolDC, but not CD8+ CD45RClow/-Treg expression itself, is donor-specific inhibitory
Example 6 patients with clinical spontaneous tolerance also show high expression of HLA-DR + CD8 + Treg expression
Survival for more than 1 year without any immunosuppressant, and maintenance of normal liver function, considered clinically actionable tolerance. We found in clinical practice that 3 cases of liver transplantation due to end-stage cirrhosis were tolerated by the clinical practice described above, and we observed HLA-DR in peripheral blood in a large number of long-lived patients (> 3 years) as measured by flow cytometry + CD8 + Treg, namely MHC-II + CD 8+ Tregs were at high expression levels (fig. 10 a). These results were significantly higher than those of acute rejection patients (B of fig. 10). These all suggest that in the course of immune tolerance, HLA-DR + CD8 + Tregs play an important role, and may be molecular markers for which immune tolerance is prone to develop.
The above examples are merely illustrative of the preferred embodiments of the present invention and the scope of the present invention is not limited thereto. Any modification, equivalent replacement or change made on the basis of the present invention by those skilled in the art is within the protection scope of the present invention.
Claims (10)
1. A method of inducing transplant immune tolerance comprising administering mTOR-targeted tolerogenic dendritic cells (Rapa-toldcs) to a transplanted mammal using a pre-operative pre-stimulation and post-operative reinfusion protocol.
2. The method of claim 1, wherein the method is performed using a pre-operative pre-stimulation plus 1 to 3 reinfusion cycles within 1 month post-operative.
3. The method of claim 1, wherein the method is performed using a pre-operative 7 day pre-stimulation + day post-operative reinfusion protocol, or wherein the method is performed using a pre-operative 7 day pre-stimulation + day 7 post-operative reinfusion protocol.
4. The method of claim 1, wherein the method is performed using a pre-stimulation regimen of 7 days prior to surgery plus a reinfusion regimen of 7, 14 and 28 days post surgery.
5. The method according to any one of claims 1 to 4, wherein when the mammal is a rat, the effective number of cells per reinfusion is 1 x 10, depending on the weight of the rat and the surgical practice 6 -1×10 7 。
6. The method of any one of claims 1 to 4, wherein the Rapa-tolDCs induce high expression of MHC-II in the transplant + CD8 of (1) + CD45RC low/- Generation of tregs.
7. The method of any one of claims 1 to 4, wherein the transplantation comprises cell transplantation, organ transplantation, or tissue transplantation; wherein the cell transplantation comprises stem cell transplantation, regulatory cell transplantation, islet cell transplantation or effector cell transplantation; organ transplantation includes kidney transplantation, liver transplantation, heart transplantation, small intestine transplantation, lung transplantation, pancreas transplantation, or combined organ transplantation; tissue transplantation includes corneal transplantation, limb transplantation or face transplantation, preferably liver transplantation;
also preferably, the mammal is a rat, mouse, pig, rabbit, dog, cat, horse, cow, sheep, monkey, chimpanzee, or human.
8. The method according to any one of claims 1 to 4, wherein the Rapa-tolDC is prepared by a process comprising:
taking bone marrow mesenchymal stem cells or peripheral blood PMBC, carrying out adherent culture by using a phenol red-free culture solution containing 1% -10% FBS to obtain precursor cells of the DC, and adding a complete culture medium containing GM-CSF and IL-4; culturing for full change on day 2, half change on day 4, supplementing cell factors GM-CSF and IL-4, maintaining concentration, and adding rapamycin when changing liquid on day 2 and day 4; culturing to day 6, half-changing the culture medium, supplementing GM-CSF to maintain the concentration constant, and adding LPS, and continuing culturing, preferably,
the concentration of GM-CSF is 1-40ng/ml, the concentration of IL-4 is 0.5-20ng/ml, the concentration of rapamycin is 1-40nM, the concentration of LPS is 0.01-1. Mu.g/ml, optionally,
the concentration ratio of GM-CSF to IL-4 is 2:1; rapamycin was added at 10nM concentrations at day 2 and day 4 changes, respectively, and LPS was added at 0.05. Mu.g/mL at day 6.
9. Use of Rapa-tolDC as defined in any one of claims 1 to 7 in the preparation of an animal model of transplant tolerance.
10. An animal model of transplant tolerance prepared by the method of any one of claims 1 to 7.
Priority Applications (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310134489.0A CN115778984A (en) | 2023-02-20 | 2023-02-20 | Method and application for inducing transplantation immune tolerance and transplantation tolerance animal model |
| CN202310939356.0A CN116999461A (en) | 2023-02-20 | 2023-02-20 | HLA-DR + CD8 + Application of Treg in preparation of kit or medicine |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310134489.0A CN115778984A (en) | 2023-02-20 | 2023-02-20 | Method and application for inducing transplantation immune tolerance and transplantation tolerance animal model |
Related Child Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202310939356.0A Division CN116999461A (en) | 2023-02-20 | 2023-02-20 | HLA-DR + CD8 + Application of Treg in preparation of kit or medicine |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN115778984A true CN115778984A (en) | 2023-03-14 |
Family
ID=85431018
Family Applications (2)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202310134489.0A Pending CN115778984A (en) | 2023-02-20 | 2023-02-20 | Method and application for inducing transplantation immune tolerance and transplantation tolerance animal model |
| CN202310939356.0A Pending CN116999461A (en) | 2023-02-20 | 2023-02-20 | HLA-DR + CD8 + Application of Treg in preparation of kit or medicine |
Family Applications After (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202310939356.0A Pending CN116999461A (en) | 2023-02-20 | 2023-02-20 | HLA-DR + CD8 + Application of Treg in preparation of kit or medicine |
Country Status (1)
| Country | Link |
|---|---|
| CN (2) | CN115778984A (en) |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1367688A (en) * | 1999-04-14 | 2002-09-04 | 美国政府卫生与公众服务部 | Compositions containing immunotoxins and agents that inhibit dendritic cell maturation for inducing immune tolerance to graft |
| US20210371820A1 (en) * | 2017-12-01 | 2021-12-02 | Nanjing Airui Biological Technology Co., LTD | Method for amplifying antigen-specific regulatory t cells in vitro |
| CN113826014A (en) * | 2019-04-16 | 2021-12-21 | 明尼苏达大学董事会 | Biomarkers of transplant tolerance induced by apoptotic donor leukocytes |
| CN115044553A (en) * | 2022-08-16 | 2022-09-13 | 首都医科大学附属北京朝阳医院 | mTOR-targeted tolerant dendritic cell and preparation method and application thereof |
-
2023
- 2023-02-20 CN CN202310134489.0A patent/CN115778984A/en active Pending
- 2023-02-20 CN CN202310939356.0A patent/CN116999461A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1367688A (en) * | 1999-04-14 | 2002-09-04 | 美国政府卫生与公众服务部 | Compositions containing immunotoxins and agents that inhibit dendritic cell maturation for inducing immune tolerance to graft |
| US20210371820A1 (en) * | 2017-12-01 | 2021-12-02 | Nanjing Airui Biological Technology Co., LTD | Method for amplifying antigen-specific regulatory t cells in vitro |
| CN113826014A (en) * | 2019-04-16 | 2021-12-21 | 明尼苏达大学董事会 | Biomarkers of transplant tolerance induced by apoptotic donor leukocytes |
| CN115044553A (en) * | 2022-08-16 | 2022-09-13 | 首都医科大学附属北京朝阳医院 | mTOR-targeted tolerant dendritic cell and preparation method and application thereof |
Non-Patent Citations (1)
| Title |
|---|
| 张升宁;李立;: "未成熟树突状细胞与肝脏移植免疫耐受" * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN116999461A (en) | 2023-11-07 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| JP4740452B2 (en) | Methods, compositions and methods of using mesenchymal stem cells for the prevention and treatment of immune responses in transplantation | |
| EP2084267B1 (en) | Cancer stem cell antigen vaccines and methods | |
| EP0812201B1 (en) | In vitro amplification of stem cells | |
| JP3201610B2 (en) | How to treat a tumor | |
| JP6422344B2 (en) | Methods for increasing allogeneic antigen-reactive regulatory T cells | |
| Lai et al. | Mouse embryonic stem cell-derived thymic epithelial cell progenitors enhance T-cell reconstitution after allogeneic bone marrow transplantation | |
| Gombert et al. | Fibroblasts prevent apoptosis of IL‐2‐deprived T cells without inducing proliferation: a selective effect on Bcl‐XL expression | |
| EA023106B1 (en) | Method for producing partially matured dendritic cells having increased expression of interleukin il-12 | |
| KR20100033471A (en) | Human cancer stem cells | |
| US20190183995A1 (en) | Method of expanding nk cell and composition for culturing | |
| Li et al. | Effects of bone marrow mesenchymal stem cells on hematopoietic recovery and acute graft-versus-host disease in murine allogeneic umbilical cord blood transplantation model | |
| CN108893443A (en) | A kind of Efficient amplification method of cytokine induction umbilical cord blood natural killer | |
| WO2020038299A1 (en) | Exosome-based antitumor vaccine | |
| JP2015513403A5 (en) | ||
| JP2016510756A (en) | Individual high-purity hepatocellular carcinoma stem cells, preparation method thereof and use thereof | |
| CN112608896A (en) | NK cell culture method and application thereof | |
| CN106554942A (en) | A kind of efficient clinical grade CD56+The preparation method of group's immunocyte | |
| Zhao et al. | Upregulation of heme oxygenase-1 endues immature dendritic cells with more potent and durable immunoregulatory properties and promotes engraftment in a stringent mouse cardiac allotransplant model | |
| US20210238549A1 (en) | Use of memory lymphocyte population in liver cancer treatment | |
| JPH089967A (en) | Induced culture method for cytotoxic t lymphocyte against cancer cell | |
| Ye et al. | B7-H1 expression is required for human endometrial regenerative cells in the prevention of transplant vasculopathy in mice | |
| CN115044553B (en) | mTOR-targeted tolerant dendritic cell and preparation method and application thereof | |
| Lin et al. | Arsenic trioxide is a novel agent for combination therapy to prolong heart allograft survival in allo-primed T cells transferred mice | |
| CN115778984A (en) | Method and application for inducing transplantation immune tolerance and transplantation tolerance animal model | |
| CN108690829B (en) | Method for efficiently amplifying NK cells |
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 | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20230314 |