CN1463593A - Process for preparing type C hepatitis virus natural infestation mice model and use thereof - Google Patents
Process for preparing type C hepatitis virus natural infestation mice model and use thereof Download PDFInfo
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- CN1463593A CN1463593A CN 02112096 CN02112096A CN1463593A CN 1463593 A CN1463593 A CN 1463593A CN 02112096 CN02112096 CN 02112096 CN 02112096 A CN02112096 A CN 02112096A CN 1463593 A CN1463593 A CN 1463593A
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- liver cell
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- 238000010172 mouse model Methods 0.000 title claims abstract description 24
- 206010061217 Infestation Diseases 0.000 title claims description 10
- 241000700605 Viruses Species 0.000 title claims description 10
- 208000006454 hepatitis Diseases 0.000 title claims description 10
- 231100000283 hepatitis Toxicity 0.000 title claims description 10
- 238000004519 manufacturing process Methods 0.000 title 1
- 210000005229 liver cell Anatomy 0.000 claims abstract description 28
- 208000005176 Hepatitis C Diseases 0.000 claims abstract description 19
- 238000000034 method Methods 0.000 claims abstract description 12
- 238000011160 research Methods 0.000 claims abstract description 12
- 239000003814 drug Substances 0.000 claims abstract description 9
- 230000000694 effects Effects 0.000 claims abstract description 7
- 241001465754 Metazoa Species 0.000 claims abstract description 6
- 241000699666 Mus <mouse, genus> Species 0.000 claims description 38
- 241000711549 Hepacivirus C Species 0.000 claims description 28
- 208000015181 infectious disease Diseases 0.000 claims description 16
- 230000006058 immune tolerance Effects 0.000 claims description 12
- 210000004185 liver Anatomy 0.000 claims description 10
- 206010067125 Liver injury Diseases 0.000 claims description 8
- 230000002440 hepatic effect Effects 0.000 claims description 8
- 231100000753 hepatic injury Toxicity 0.000 claims description 8
- 238000002360 preparation method Methods 0.000 claims description 8
- 210000002966 serum Anatomy 0.000 claims description 7
- 229940079593 drug Drugs 0.000 claims description 4
- 210000003494 hepatocyte Anatomy 0.000 claims description 4
- 210000000987 immune system Anatomy 0.000 claims description 4
- 230000008506 pathogenesis Effects 0.000 claims description 4
- 238000002054 transplantation Methods 0.000 claims description 4
- 230000001506 immunosuppresive effect Effects 0.000 claims description 3
- 230000036285 pathological change Effects 0.000 claims description 3
- 231100000915 pathological change Toxicity 0.000 claims description 3
- 241000699670 Mus sp. Species 0.000 claims description 2
- 210000004369 blood Anatomy 0.000 claims description 2
- 239000008280 blood Substances 0.000 claims description 2
- 230000007969 cellular immunity Effects 0.000 claims description 2
- 208000010710 hepatitis C virus infection Diseases 0.000 claims description 2
- 230000001717 pathogenic effect Effects 0.000 claims description 2
- 230000008807 pathological lesion Effects 0.000 claims description 2
- 238000011084 recovery Methods 0.000 claims description 2
- 238000012216 screening Methods 0.000 claims description 2
- MSWZFWKMSRAUBD-GASJEMHNSA-N 2-amino-2-deoxy-D-galactopyranose Chemical compound N[C@H]1C(O)O[C@H](CO)[C@H](O)[C@@H]1O MSWZFWKMSRAUBD-GASJEMHNSA-N 0.000 claims 1
- MSWZFWKMSRAUBD-UHFFFAOYSA-N beta-D-galactosamine Natural products NC1C(O)OC(CO)C(O)C1O MSWZFWKMSRAUBD-UHFFFAOYSA-N 0.000 claims 1
- 230000001575 pathological effect Effects 0.000 abstract 2
- 238000011156 evaluation Methods 0.000 abstract 1
- 238000002474 experimental method Methods 0.000 abstract 1
- 230000001900 immune effect Effects 0.000 abstract 1
- 230000008629 immune suppression Effects 0.000 abstract 1
- 238000010171 animal model Methods 0.000 description 5
- 238000002347 injection Methods 0.000 description 5
- 239000007924 injection Substances 0.000 description 5
- 102000008100 Human Serum Albumin Human genes 0.000 description 4
- 108091006905 Human Serum Albumin Proteins 0.000 description 4
- 238000007799 mixed lymphocyte reaction assay Methods 0.000 description 4
- 108090000623 proteins and genes Proteins 0.000 description 4
- 210000000952 spleen Anatomy 0.000 description 4
- 241000699660 Mus musculus Species 0.000 description 3
- 238000011830 transgenic mouse model Methods 0.000 description 3
- LCTORNIWLGOBPB-SVZMEOIVSA-N (3r,4s,5r,6r)-2-amino-6-(hydroxymethyl)oxane-2,3,4,5-tetrol Chemical compound NC1(O)O[C@H](CO)[C@H](O)[C@H](O)[C@H]1O LCTORNIWLGOBPB-SVZMEOIVSA-N 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 208000019423 liver disease Diseases 0.000 description 2
- 241000415078 Anemone hepatica Species 0.000 description 1
- 241000282577 Pan troglodytes Species 0.000 description 1
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 1
- 108700019146 Transgenes Proteins 0.000 description 1
- 231100000749 chronicity Toxicity 0.000 description 1
- 239000002299 complementary DNA Substances 0.000 description 1
- 241001493065 dsRNA viruses Species 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
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- 201000007270 liver cancer Diseases 0.000 description 1
- 208000014018 liver neoplasm Diseases 0.000 description 1
- 230000007170 pathology Effects 0.000 description 1
- 230000008929 regeneration Effects 0.000 description 1
- 238000011069 regeneration method Methods 0.000 description 1
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- 210000001519 tissue Anatomy 0.000 description 1
- 210000003462 vein Anatomy 0.000 description 1
- 230000009385 viral infection Effects 0.000 description 1
- 210000001835 viscera Anatomy 0.000 description 1
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- Investigating Or Analysing Biological Materials (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
Human liver cell is transplanted into the body of mouse to establish naturally infected mouse model of hepatitis C. The said model is superior to available models in that the experiment animal is in normal immunological state, rather than immune suppression state, and can simulate naturally human HCV infecting process for the research of the pathological mechanism and pathological damage and ultimate evaluation of treating effect of medicine for hepatitis C.
Description
Technical field
The invention belongs to biotechnology, relate to biological model,, be specifically related to the preparation and the purposes of type C hepatitis virus natural infestation mice model especially about the virus infections small animal model.
Background technology
Hepatitis C is widely current in the whole world, and the patient more than 80% can cause chronicity after being infected by hepatitis C virus (HCV), and closely related with the generation of liver cancer, and how effectively preventing and treat hepatitis C is problem demanding prompt solution.
But up to research the achieving no breakthrough property progress yet of hepatitis C today, key factor is to lack ideal animal model.Except the mankind, has only chimpanzee to the hepatitis C virus susceptible.Because experimental expenses is extremely expensive, the whole world has only the several laboratories of only a few to study.Chinese scholars is devoted to the research that the hepatitis C small animal model is set up.
Hepatitis C small animal model research at most, the most detailed should belong to transgenic mice.The transgenic mice of the multiple HCV of being integrated with different genes fragment has successively been set up in many laboratories, the gene order and each the section various combination mode that have comprised each sections such as HCV C district, E district, NS district, even the transgene mouse model of HCV cDNA full length sequence.Change the hepatitis c virus gene mouse and provide fabulous model for the immune tolerance Study on Mechanism, but for research hepatitis C pathogenesis, how infected pathological change that host's liver after the hepatitis C etc. organizes internal organs, and the aspects such as assessment of medicine curative effect, transgenic mice is powerless.At first because mouse is not the suitable host of HCV, after HCV changes mouse over to, can detect the HCV rna replicon in the mouse body, but not have the hepatic pathology infringement, different with the variation behind the human infection HCV, secondly, inject the HCV gene in mouse fertilized egg, postnatal mouse is in immune tolerance state, and is different with the natural infection process, therefore change the pathogenesis that the HCV genetic animal can not reflect hepatitis C, can not be used to screen the also curative effect of estimating anti HCV medicine.
Summary of the invention
The objective of the invention is human liver cell is implanted in the mouse body, set up mouse hepatitis C natural infection model, overcome in the past the shortcoming that when zoopery animal all is in immunosuppressive condition, has normal immune system, model contains a large amount of human liver cells, can anthropomorphic dummy HCV natural infection process, study its pathogenesis, pathological lesion, and can finally estimate anti-hepatitis C curative effect of medication, set up mouse type C hepatitis virus natural infestation model and specifically be divided into four steps:
(1) sets up the immune tolerance mouse model;
(2) set up people mouse mosaic type hepatic model;
(3) set up mouse fulminant liver injury model;
(4) set up type C hepatitis virus natural infestation mice model.
The preparation of the type C hepatitis virus natural infestation mice model that purpose of the present invention relates to realizes by following scheme: (1) with the mouse inbred lines in the human liver cell immunity birth 24 hours, only sets up the human liver cell immune tolerance and the normal mouse of whole immune system-immune tolerance mouse model.(2) human liver cell is imported the immune tolerance mouse through the hepatocyte transplantation method, set up human liver cell and mouse liver cell allophenic mice-people mouse mosaic type hepatic model; (3) injection of d-galactose amine is set up mouse fulminant liver injury model; (4) human liver cell is rebuild hepatic tissue through the input of hepatocyte transplantation method, obtain to contain the more mouse of human liver cell, blood transfusion HCV infection, the mouse model of preparation HCV natural infection.
Two distinguishing features of the present invention are: 1, with small animal model research HCV natural infection, the normal immunologic function of small animal is necessary.Key of the present invention is only to set up the human liver cell immune tolerance and the normal mouse model of whole immune system, the natural process that can anthropomorphic dummy HCV infects.2, in human liver cell and mouse liver cell allophenic mice, obtain human liver cell as much as possible, pathological change that can anthropomorphic dummy's liver, thus make this model be applicable to the research of HCV.
Embodiment
The preparation of embodiment 1 type C hepatitis virus natural infestation mice model:
(1) sets up the immune tolerance mouse model: the Balb/c mouse that newly is born be born lumbar injection normal liver cell (100 μ l, 10 in back 24 hours
5), be contrast (100 μ l) with the injecting normal saline, observe human albumin content in liver and the serum after 1 week, whether spleen weight, and mixed lymphocyte reaction (MLP) are set up with check immune tolerance model.
(2) set up people mouse mosaic type hepatic model: the 4th week is through spleen injection normal liver cell 10 μ l, 10
5, the 5th week was observed human albumin content in liver and the serum, and whether spleen weight, and mixed lymphocyte reaction (MLP) are set up with identifier mouse mosaic type hepatic model.And in the outer lateral lobe of the 6th week excision liver, the 10th week in week the back put to death, observe neonatal liver leaf disease reason section human albumin SABC, the human liver cell of check transplanting has or not regeneration function.
(3) set up mouse fulminant liver injury model: the 6th week preparation fulminant liver injury model, injection of d-galactose amine, with hepatic pathology section and Serum ALT levels, whether check mouse fulminant liver injury model is set up.
(4) set up HCV natural infection mouse model, the 7th week was injected 10 μ l, 10 through spleen
7Human liver cell, the content of human liver cell in the human albumin content check mouse people mouse mosaic type liver in observation liver and the serum.The 10th all tail vein injection HCV positive serums infect 1 week of back, and in 2 weeks, in 3 weeks, in 4 weeks, in 5 weeks, in 6 weeks, in 7 weeks, (whether check HCV natural infection mouse model is set up for IgM, IgG) level for HCV RNA and HCV antibody in 8 all observation livers continuously and the serum.
The application of embodiment 2 type C hepatitis virus natural infestation mice models
Mouse model with the present invention's foundation, animal all is in the shortcoming of immunosuppressive condition when having overcome zoopery in the past, the natural process that can anthropomorphic dummy HCV infects can be used for that the pathogenetic research of hepatitis C virus, hepatitis C pathology change research, researchs such as recovery from hepatitis C virus infection, the anti-hepatitis C medicine of screening, curative effect of medication examination are observed, observed to hepatitis C course of disease progress.
Need not further to elaborate, believe and adopt the disclosed content in front, those skilled in the art can use the present invention to greatest extent.Therefore, the preferred specific embodiments of front is interpreted as only illustrating, but not limits the scope of the invention by any way.
Claims (6)
1. the preparation of a type C hepatitis virus natural infestation mice model and purposes, it is characterized in that: human liver cell is implanted in the mouse body, set up mouse hepatitis C natural infection model, overcome in the past the shortcoming that when zoopery animal all is in immunosuppressive condition, has normal immune system, model contains a large amount of human liver cells, can anthropomorphic dummy HCV natural infection process, study its pathogenesis, pathological lesion, and can finally estimate anti-hepatitis C curative effect of medication, specifically be divided into four steps:
(1) sets up the immune tolerance mouse model;
(2) set up people mouse mosaic type hepatic model;
(3) set up mouse fulminant liver injury model;
(4) set up HCV natural infection mouse model.
2. the immune tolerance mouse model of setting up according to claim 1 is characterized in that with the mouse inbred lines in the human liver cell immunity birth 24 hours, only sets up the human liver cell immune tolerance and the normal mouse of whole immune system.
3. the people mouse mosaic type hepatic model of setting up according to claim 1 is characterized in that human liver cell is imported the immune tolerance mouse through the hepatocyte transplantation method, sets up human liver cell and mouse liver cell allophenic mice;
4. the mouse fulminant liver injury model of setting up according to claim 1 is characterized in that injecting D-galactosamine, sets up mouse fulminant liver injury model.
5. the HCV natural infection mouse model of setting up according to claim 1, it is characterized in that human liver cell is rebuild hepatic tissue through the input of hepatocyte transplantation method, acquisition contains the more mouse of human liver cell, the blood transfusion HCV infection, observe HCV RNA and HCV antibody (IgM in liver and the serum, IgG) level, the mouse model of preparation HCV natural infection.
6. the purposes of type C hepatitis virus natural infestation mice model according to claim 1, it is characterized in that the natural process that can anthropomorphic dummy HCV infects, be used to study the natural infection process of hepatitis C, pathological change that can anthropomorphic dummy's liver can be used for the pathogenetic research of hepatitis C virus, observes recovery from hepatitis C virus infection, researchs such as the anti-hepatitis C medicine of screening, curative effect of medication examination.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB02112096XA CN1189076C (en) | 2002-06-15 | 2002-06-15 | Process for preparing type C hepatitis virus natural infestation mice model and use thereof |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB02112096XA CN1189076C (en) | 2002-06-15 | 2002-06-15 | Process for preparing type C hepatitis virus natural infestation mice model and use thereof |
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| Publication Number | Publication Date |
|---|---|
| CN1463593A true CN1463593A (en) | 2003-12-31 |
| CN1189076C CN1189076C (en) | 2005-02-16 |
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| Application Number | Title | Priority Date | Filing Date |
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| CNB02112096XA Expired - Fee Related CN1189076C (en) | 2002-06-15 | 2002-06-15 | Process for preparing type C hepatitis virus natural infestation mice model and use thereof |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101864397A (en) * | 2009-04-14 | 2010-10-20 | 中国医学科学院基础医学研究所 | Establishment of in vitro cell model supporting HCV 1b sub-genome duplication |
| CN102281758A (en) * | 2009-01-16 | 2011-12-14 | 公益财团法人实验动物中央研究所 | Mouse having human hepatocytes transplanted therein |
| CN103920166A (en) * | 2013-01-10 | 2014-07-16 | 中国计量学院 | Establishment method and application of novel hepatitis B animal model |
| CN107156059A (en) * | 2017-03-07 | 2017-09-15 | 浙江大学 | A kind of method of utilization stem cell constructing humanization chronic hepatitis B mouse model |
-
2002
- 2002-06-15 CN CNB02112096XA patent/CN1189076C/en not_active Expired - Fee Related
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102281758A (en) * | 2009-01-16 | 2011-12-14 | 公益财团法人实验动物中央研究所 | Mouse having human hepatocytes transplanted therein |
| CN101864397A (en) * | 2009-04-14 | 2010-10-20 | 中国医学科学院基础医学研究所 | Establishment of in vitro cell model supporting HCV 1b sub-genome duplication |
| CN103920166A (en) * | 2013-01-10 | 2014-07-16 | 中国计量学院 | Establishment method and application of novel hepatitis B animal model |
| CN107156059A (en) * | 2017-03-07 | 2017-09-15 | 浙江大学 | A kind of method of utilization stem cell constructing humanization chronic hepatitis B mouse model |
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| Publication number | Publication date |
|---|---|
| CN1189076C (en) | 2005-02-16 |
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