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A kind of nmosd animal model construction method

A technology for building methods and animal models, applied in animal husbandry and other directions, can solve problems such as high precision requirements, failure to simulate disease characteristics, and low success rate

Active Publication Date: 2021-10-08
管阳太
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Problems solved by technology

However, the disease site of this model is limited to the injection area, and there is no observational evidence of the optic nerve and spinal cord. This model fails to simulate the disease characteristics of the optic nerve and spinal cord in the course of NMOSD.
[0007] 3) NMOSD model induced by AQP4-reactive T cells: This model confirms the potential role of AQP4-reactive T cells in causing damage to central oligodendrocytes in behavioral disorders, which is conducive to trying specific immunotherapy against AQP4 antigens, but the operation steps are relatively complex. Many, high precision requirements, low success rate
[0008] Taken together, there is currently no universally accepted animal model of NMOSD that addresses more questions

Method used

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  • A kind of nmosd animal model construction method

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Embodiment 1

[0027] see figure 1 , a method for constructing an NMOSD animal model, comprising the steps of:

[0028] S1. Establishing a mouse EAE model;

[0029] S2. Extracting patient NMO-IgG and normal human serum complement;

[0030] S3. Opening the blood-brain barrier of EAE model mice: at the peak of the onset of EAE in mice, the mice were anesthetized, and the top of the skull was depilated; sonovel microbubbles were injected through the tail vein of the mice as a contrast agent, and a low-frequency focused ultrasound probe was used to radiate through the skull. Irradiate the head of the mouse for 1 minute;

[0031] S4. Injecting human NMO-IgG and complement to EAE model mice: Immediately after irradiation, each EAE model mouse was injected with 100 μg NMO-IgG and 100 μl complement through the tail vein.

[0032] In this example, the mouse EAE model was induced by neuromyelitis optica IgG antibody and complement immunization, and the blood-brain barrier was opened by using low-fr...

Embodiment 2

[0036] S1. Establish mouse EAE model:

[0037] 1) Preparation of MOG antigen dilution: Weigh an appropriate amount of MOG 35～55 Peptide freeze-dried powder is dissolved in phosphate buffer solution to make the concentration 2mg / ml;

[0038] 2) Preparation of complete Freund's adjuvant: take an appropriate amount of incomplete Freund's adjuvant, add inactivated Mycobacterium tuberculosis to make the concentration reach 4mg / ml;

[0039] 3) Preparation of antigen emulsion: mix equal volumes of MOG antigen diluent and complete Freund's adjuvant, and fully emulsify on ice to a water-in-oil state. At this time, MOG 35～55 The concentration is 1mg / ml, and the concentration of inactivated Mycobacterium tuberculosis is 2mg / ml;

[0040] 4) Preparation of EAE animal model: mice were anesthetized, 200 μL of antigen emulsion was subcutaneously injected at four points on the back of the mice, and 200 ng of pertussis toxin was injected intraperitoneally into each mouse on the day of immunizat...

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Abstract

The invention relates to the technical field of animal models, and discloses a method for constructing an NMOSD animal model, including preparing a mouse EAE model, extracting human NMO-IgG and complement, using low-frequency focused ultrasound combined with microbubble technology to open the blood-brain barrier, and injecting human NMO -IgG and complement, so that NMO-IgG enters the central nervous system through the blood-brain barrier, and establishes an animal model of neuromyelitis optica spectrum disease. The present invention adopts neuromyelitis optica IgG antibody and complement immune induction, and uses physical methods to open the blood-brain barrier non-invasively, temporarily and reversibly, so that NMO-IgG and complement can enter the animal center to induce neuromyelitis optica spectrum diseases, thereby establishing optic neurospinal cord Animal models of inflammatory spectrum diseases; compared with other existing animal models, the present invention better simulates the pathogenesis of human-related diseases, more objectively reflects the pathological process and pathological characteristics of NMOSD, and provides new research for related research platform.

Description

technical field [0001] The invention relates to the technical field of animal models, in particular to a method for constructing an NMOSD animal model. Background technique [0002] Neuromyelitis optica spectrum disease (NMOSD) is one of the most rapidly updated and advanced diseases in the field of neurology in the past decade. NMOSD is a type of acute or subacute autoimmune demyelinating disease in which the optic nerve and spinal cord are involved simultaneously or sequentially. It seriously threatens human health due to its high disability rate, easy recurrence, and poor prognosis. Although NMOSD is different from optic neuritis and multiple sclerosis (MS) in terms of clinical symptoms, course of disease, affected parts, laboratory tests and responses to various treatments, there are great similarities. Continuity and overlap, it is difficult to distinguish NMOSD from diseases such as MS only relying on the above characteristics. [0003] In 2004, Lemmon and his collea...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01K67/02
CPCA01K67/02A01K2207/10A01K2227/105A01K2267/03
Inventor 管阳太罗佳莹谢冲朱德生韩露王亦舒丁婕洪荣华杨笑白书维郝勇刘明媛
Owner 管阳太
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