CN104758080A - Method for establishing non-human primate autoimmune cerebrospinal meningitis model and application of model - Google Patents
Method for establishing non-human primate autoimmune cerebrospinal meningitis model and application of model Download PDFInfo
- Publication number
- CN104758080A CN104758080A CN201510128956.4A CN201510128956A CN104758080A CN 104758080 A CN104758080 A CN 104758080A CN 201510128956 A CN201510128956 A CN 201510128956A CN 104758080 A CN104758080 A CN 104758080A
- Authority
- CN
- China
- Prior art keywords
- model
- emulsion
- machin
- cerebrospinal meningitis
- injection
- 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
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61D—VETERINARY INSTRUMENTS, IMPLEMENTS, TOOLS, OR METHODS
- A61D7/00—Devices or methods for introducing solid, liquid, or gaseous remedies or other materials into or onto the bodies of animals
Abstract
The invention discloses a method for establishing a non-human primate autoimmune cerebrospinal meningitis model and the application of the model. The specific technical scheme comprises the following steps; a machin is selected as the non-human primate experimental animals; MOG34-56(100Mug/ml) is prepared into an emulsion (MOG CFA=1; 1); after the experimental machin is anesthetized, the machin is intradermally injected with 1ml of prepared emulsion at 10 injection points; on the seventh day after the first injection, an MOG-CFA emulsion is prepared by use of the same method, and the second immune injection is performed at the same dosage and by use of the same method; the experimental autoimmune cerebrospinal meningitis model established by use of the method has the clinical characters of remission-relapse, and can be widely used to the related field of disseminated sclerosis diseases. Compared with the existing rhesus method and model, the method and the model also have the characteristics of short induction period, low cost, rich monkey resources and the like, and also have the application value which cannot be realized by rodent models.
Description
Technical field
The present invention relates to the foundation of non-human primate disease model, be specifically related to a kind of non-human primates autoimmune cerebrospinal meningitis method for establishing model and application.
Background technology
Experimental Autoimmune encephalomyelitis (experimental autoimmune encephalomyelitis, EAE) be by isotype, allotype, xenogenesis abnormal shape causes encephalitis antigen induction, experiment Sensitivity animal produces, mediated by cell immune response, with central nervous system (central nervous system, CNS) white matter demyelination is the delayed allergy autoimmune disease of feature, its immunopathogenesis and disease damage and human multiple sclerosis (multiple sclerosis, MS) similar, be acknowledged as the ideal model of research multiple sclerosis disease.
Current Experimental Autoimmune encephalomyelitis (EAE) model is normally based upon on rodent.Because Rodents is far apart with the mankind on evolutionary relationship, practice shows, the experimental result utilizing rodent Experimental Autoimmune encephalomyelitis (EAE) model to produce is difficult to be verified in people's experiment, the development-success ratio of assessment medicine is lower, little to practical guided significance with the experimental data that it obtains for object of study.Each system physiological function such as anatomical features, immunity of non-human primates and very close with the mankind to reactivity of disease and medicine etc. carries out the research of immunology relevant disease and treatment and the Sensitivity animal of effectiveness and safety evaluatio before carrying out clinical drug.
Along with the development of new drug development technology, the especially maturation of macromolecular drug (as antibody, nucleic acid, protein medicaments) technology, drug specificity action target spot and be the successful important factor in order of medicament research and development with test model cross reactivity.The relationship of non-human primate and human body is close, and the structure of target spot is similar with biosystem, has good cross reactivity.Therefore set up non-human primates Experimental Autoimmune encephalomyelitis (EAE) model and the data produced in the urgent need to, model meeting Treatment of Multiple Sclerosis drug evaluation are had important directive significance to the research of multiple sclerosis and to practical applications such as treatments.
MOG is a kind of expression at the transmembrane glycoprotein on maincenter oligodendrocyte and myelin not surface, and content seldom but has very high immunogenicity in vivo.Research shows, MOG uniquely can cause demyelination antibody response, can cause again central nervous system's myelin protein composition of t cell responses.Therefore, generation experimental autoimmune encephalomyelitis (EAE) was originally induced can better to help to illustrate the pathogenesis of multiple sclerosis (MS) with MOG as immunity.
Herbert P.M.Brok etc. once reported the model using MOG to induce Rhesus Macacus (Macaca mulatta) EAE, its method is: with Rhesus Macacus as laboratory animal, MOG Emulsion (MOG PBS solution: CFA=1:1) is injected in Intradermal, and after 4 weeks intradermal injection MOG Emulsion (MOG NS solution: IFA=1:1) again.But the shortcoming of this model is: 1. single morbidity; 2. abductive approach required time is long; 3. Rhesus Macacus individuality is larger; 4. adopt more subjective symptom judgment criteria.These shortcomings make this model cannot scientific research, assessing drug actions and industrialization practice in application.
Present invention utilizes machin (Macaca fascicularis) laboratory animal of a large amount of livestock on hand of China, creative use MOG (the peptide section of MOG34-56) secondary tachysynthesis method, establish machin laboratory animal EAE model, it has the advantages that alleviation-recurrence shows repeatedly.Through kinds of experiments objective determination, the model characteristics of incidence that the present invention sets up and Pathological and human multiple sclerosis have similarity highly.
Summary of the invention
The present invention establishes a kind of method of non-human primates Experimental Autoimmune cerebrospinal meningitis (EAE) model.
The present invention is achieved by the following technical solutions:
A, non-human primate: machin
B, preparation immune induction Emulsion: MOG34-56 (100 μ g/ml) is mixed with Emulsion (MOG solution: CFA=1:1);
C, first time inoculation: the Emulsion that every machin animal injection 1ml prepares, point 10 injection site, carry out intradermal injection respectively;
D, second time inoculation: the 7th day after first time inoculation, by above-mentioned same preparation Emulsion, the method same by first time inoculation and dosage inoculation again;
The method for building up of a kind of non-human primates Experimental Autoimmune cerebrospinal meningitis model provided by the present invention, preferably: the use MOG34-56 peptide section of innovation carries out twice immunity, and interval is 7 days.
The method for building up of a kind of non-human primates Experimental Autoimmune cerebrospinal meningitis model provided by the present invention, preferably: creative use machin (Macaca fascicularis) is as laboratory animal.
EAE models applying advantage of the present invention is:
Animal model of the present invention has the using value that rodent models cannot realize:
Each system physiological function such as a, anatomical features, immunity and very close with the mankind to the reactivity of disease and medicine;
B is close with people's relationship, and the structure of target spot is similar with biosystem, for drug specificity (as antibody, nucleic acid, protein medicaments etc.) action target spot provides good cross reactivity.
Animal model of the present invention has more actual application value compared with macaque model:
The method of a, MOG34-56 peptide spacer segment twice immunity in 7 days, shortens the induction period of model;
B, morbidity have the pathogeneticing characteristic repeatedly of relapsing-remitting type, have the similarity of height with human multiple sclerosis;
C, animal individual is little compared with Rhesus Macacus, uses less test medicine, reduces costs;
D, the machin adopting monkey field to breed in a large number, genetic background is clear, and quality control is strict, experimental monkey groups aboundresources.
Non-human primate EAE model of the present invention has been widely used in the association area for multiple sclerosis disease, such as:
The research and development of a, the diagnostic techniques utilizing model to carry out being correlated with and reagent, bio-identification thing and technology and translational medicine research tool;
B, the sickness rate utilizing model, occurring degree and pathogeneticing characteristic, the various intervention techniques exploitation carrying out being correlated with and drug evaluation etc.;
C, judging quota, measuring technique and the determining instrument equipment development etc. that utilize model to carry out being correlated with;
D, the data base utilizing the animal material foundation of model to be correlated with, gene bank, material depot and large data analysis etc.
Accompanying drawing explanation
Fig. 1 is the abductive approach figure that MOG34-56 induces machin EAE
Fig. 2 is injection site and the immune effect figure of intradermal injection MOG34-56 Emulsion
Fig. 3 is the scoring figure that MOG34-56 induces machin EAE different time, the Signs progress of display morbidity-recover-recurrence
Fig. 4 is the machin cerebral lesion stove figure that magnetic resonance (MRI) axially different scanning display MOG34-56 induces EAE, its contrast, lesions showed degree and scoring relation just with clinical score.
Fig. 5 is brain tissue slice (the H & E dyes) figure that MOG34-56 induces EAE machin, the petechia (upper left corner) of display intralesional and inflammatory cell infiltration (lower right corner).
Below by way of concrete case study on implementation, the present invention is described in further detail, but do not limit the present invention, those skilled in the art can make various change and distortion according to the present invention, only otherwise depart from spirit of the present invention, all should belong to the scope of claims of the present invention.
Detailed description of the invention
Laboratory animal: 4 machins (Macaca fascicularis, male, 4 years old, body weight 3-5 kilogram);
Main agents: complete Freund's adjuvant (Complete Freund adjuvant, CFA); MOG34-56 peptide section;
The preparation of immune induction Emulsion: be dissolved in by MOG34-56 in normal saline (NS), with equal-volume CFA emulsifying 5 minutes under 4 DEG C of conditions;
Experimental technique:
By the MOG-CFA Emulsion prepared, point 10 injection site are injected in machin animal skins, every monkey injection 1ml;
And in injecting the 7th day after (immunity) first, preparing MOG-CFA Emulsion by same method, carrying out second time inoculation by same dose and method;
Experimental result:
According to after inoculation to the observation of monkey animal symptom, scoring, brain nuclear-magnetism (MRI) scanning and the means inspection such as histopathology section: all successively there is obvious multiple sclerosis symptom in the 4-8 week in week after first immunisation injection in all 4 machins, and presents the feature of morbidity-recovery-recurrence;
Model success rate: be 100%.
Claims (9)
1. set up a kind of method of non-human primates autoimmune cerebrospinal meningitis model, described method for establishing model is:
A, select machin as non-human primates laboratory animal;
B, preparation immune induction Emulsion: MOG34-56 peptide section (100 μ g/ml) is mixed with Emulsion (MOG solution: CFA=1:1);
C, first time inoculation: the Emulsion that every machin animal injection 1ml prepares, point 10 injection site, carry out intradermal injection respectively;
D, second time inoculation: the 7th day after first time inoculation, by above-mentioned same preparation Emulsion, the method same by first time inoculation and dosage inoculation again.
2. the method for claim 1, is characterized in that: the method using twice immunity in 7 days of MOG34-56 peptide spacer segment.
3. the method for claim 1, is characterized in that: use machin (Macaca fascicularis) as experimental model animal.
4. the method for claim 1, is setting up the application in non-human primates Experimental Autoimmune cerebrospinal meningitis model.
5. apply as claimed in claim 4, it is characterized in that: described animal is non-human primate, row are as macaque and Adeps seu carnis Rhiopithecus roxellanae monkey etc.
6. the method for claim 1, in the application based on non-human primates Experimental Autoimmune cerebrospinal meningitis model.
7. apply as claimed in claim 6, it is characterized in that: use the method establishment such as biology, iconography, electronics, physics, chemistry, data analysis to the detection of each phases such as model disease degree and morbidity, recovery and recurrence and assessment.
8. apply as claimed in claim 6, it is characterized in that: evaluate a kind of technology, method, compound, device in the application detected or in intervention model disease.
9. apply as claimed in claim 8, it is characterized in that: described technology, method, device are detection technique and index, biological label thing, treatment or prevention material (as compound, antibody, protein, nucleic acid, nucleoside etc.).
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510128956.4A CN104758080A (en) | 2015-03-24 | 2015-03-24 | Method for establishing non-human primate autoimmune cerebrospinal meningitis model and application of model |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510128956.4A CN104758080A (en) | 2015-03-24 | 2015-03-24 | Method for establishing non-human primate autoimmune cerebrospinal meningitis model and application of model |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN104758080A true CN104758080A (en) | 2015-07-08 |
Family
ID=53640428
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510128956.4A Pending CN104758080A (en) | 2015-03-24 | 2015-03-24 | Method for establishing non-human primate autoimmune cerebrospinal meningitis model and application of model |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104758080A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106986930A (en) * | 2017-04-14 | 2017-07-28 | 广东省生物资源应用研究所 | A kind of albumen for inducing machin experimental autoimmune encephalomyelitis animal model and application |
-
2015
- 2015-03-24 CN CN201510128956.4A patent/CN104758080A/en active Pending
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106986930A (en) * | 2017-04-14 | 2017-07-28 | 广东省生物资源应用研究所 | A kind of albumen for inducing machin experimental autoimmune encephalomyelitis animal model and application |
| CN106986930B (en) * | 2017-04-14 | 2021-06-11 | 广东省生物资源应用研究所 | Protein for inducing experimental autoimmune encephalomyelitis of cynomolgus monkey and application |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Capilla et al. | Animal models: an important tool in mycology | |
| Rouby et al. | Evidence of intrauterine transmission of lumpy skin disease virus | |
| Love | Oxytocin, motivation and the role of dopamine | |
| CN105524984A (en) | Method and equipment for neoantigen epitope prediction | |
| CN102449172A (en) | Compositions and methods for administration of vaccines against dengue virus | |
| Moosavi et al. | Intra CA1 insulin microinjection improves memory consolidation and retrieval | |
| Kivity et al. | Abnormal olfactory function demonstrated by manganese‐enhanced MRI in mice with experimental neuropsychiatric lupus | |
| Giralt et al. | Active induction of experimental autoimmune encephalomyelitis (EAE) with MOG 35–55 in the mouse | |
| Heckendorn et al. | The genetic basis for the selection of dairy goats with enhanced resistance to gastrointestinal nematodes | |
| CN104758080A (en) | Method for establishing non-human primate autoimmune cerebrospinal meningitis model and application of model | |
| Eltayeb et al. | Effector stage CC chemokine receptor-1 selective antagonism reduces multiple sclerosis-like rat disease | |
| CN107459546B (en) | A kind of tRF-Gly antisenses chain inhibitor and its application | |
| CN105561343A (en) | DNA vaccine for preventing toxoplasmosis of humans or animals | |
| CN106086232B (en) | It is a kind of for identifying duplex PCR detection primer, kit and the detection method of sheep of virus and capripox virus | |
| CN103933550A (en) | Method for establishing Macaca fascicularis experimental autoimmune encephalomyelitis model and application thereof | |
| Sindato et al. | Safety, immunogenicity and antibody persistence of rift valley fever virus clone 13 vaccine in sheep, goats and cattle in Tanzania | |
| Tian et al. | In vivo measures of nigrostriatal neuronal response to unilateral MPTP treatment | |
| WO2002026785A2 (en) | Epitopes of virus hepatitis c specifically cd4+ t lymphocytes | |
| Owens | A neuroprimer: principles of central nervous system immunity | |
| Nissen et al. | Preclinical model of multiple sclerosis: Methods in autoimmune demyelination | |
| CN110042127A (en) | Preparation, preparation method and the application of cells into cardiomyocytes delivering target nucleic acid molecules | |
| CN110038116A (en) | The purposes of people hepatic secretion Protein G PNMB or its antagonist or agonist | |
| Kitab et al. | Experimental in vitro and in vivo systems for studying the innate immune response during dengue virus infections | |
| Pujol et al. | Serologic response and adverse effects of recombinant canine distemper vaccination in three aquarium-housed walruses (Odobenus rosmarus) | |
| Gerlinger et al. | Gene Doping: Scientific Basis-Gateways-Monitoring |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20150708 |
|
| WD01 | Invention patent application deemed withdrawn after publication |