induction of experimental autoimmune encephalomyelitis in a mouse model

Induction of Experimental Autoimmune Encephalomyelitis in a Mouse Model

Begin by subcutaneously injecting 10- to 13-week-old female mice with 200 micrograms of myelin oligodendrocyte glycoprotein, emulsified in 200 microliters of complete Freund&#39;s adjuvant containing 400 micrograms of Mycobacterium tuberculosis. Immediately, and 24 hours later, intraperitoneally inject the mice with 0.2 micrograms of pertussis toxin in 200 microliters of PBS.
It is essential that mice have been adapted both to handling and the experimental environment before induction to avoid inducing stress, which can prevent the development of clinical symptoms.
One week after the injection, examine the mice daily for clinical symptoms as described in the table.

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All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.
1. Experimental autoimmune encephalomyelitis or EAE Model
<ol>
	<li>Induction of the EAE model
	<ol>
		<li>Use 10- to 13-week-old female 129S4/SvJae&#215;C57BL/6 mice for the induction of EAE.</li>
		<li>Give the mice a subcutaneous injection, into the upper and lower back, of the encephalitogenic MOG35 - 55&#160;(myelin oligodendrocyte glycoprotein) peptide (200 &#181;g), emulsified in 200 &#181;l complete Freund`s adjuvant (CFA) containing 400 &#181;g&#160;Mycobacterium tuberculosis. 
		NOTE: As a negative, non-disease-inducing sham control, incomplete Freund&#39;s adjuvant containing&#160;Mycobacterium tuberculosis, in the same volume and by the same route, can be used.</li>
		<li>Thereafter, and again 24 hr later, give the mice an intraperitoneal injection of pertussis toxin (in total 0.2 &#181;g, diluted in 200 &#181;l phosphate-buffered saline, PBS). Use untreated mice as the control group, to be able to compare diseased with healthy animals.&#160;&#160;&#160;&#160;&#160;&#160;&#160; 
		NOTE: It is also possible to induce EAE with 200 - 300 &#181;g MOG35 - 55&#160;peptide, 300 - 500 &#181;g of&#160;Mycobacterium tuberculosis&#160;in the CFA, and 0.2 - 0.3 &#181;g pertussis toxin in a volume of 0.1 - 0.2 ml per injection.</li>
		<li>Beginning one week after the injection, examine mice daily for clinical symptoms (see step 1.2.1)&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160; 
		NOTE: The day of disease onset varies in different experiments, but under the conditions in our laboratory, this is around day 11 and thus, here day 14 is defined as 3 days after disease onset. All mice in the present study developed clinical symptoms.</li>
	</ol>
	</li>
</ol>
2. Scoring of the mice
<ol>
	<li>Classify clinical symptoms by clinical scores as follows: 0) no signs, 0.5) distal paralysis of the tail, 1) complete paralysis of the tail, 1.5) limp tail and mild weakness of hind legs, 2) limp tail and severe weakness of hind legs, 2.5) limp tail and paralysis of one hind leg, 3) limp tail and paralysis of both hind legs, 3.5) paralysis of both hind limbs and weakness of one fore limb (mice achieving this score were euthanized, in keeping with local ethical guidelines).</li>
</ol>

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>EAE Kit</td>
			<td>Hooke Laboratories, Lawrence, USA</td>
			<td>EK2110</td>
			<td></td>
		</tr>
	</tbody>
</table>

Source: Barthelmes, J. et al., <a href="https://app.jove.com/v/53933">Induction of Experimental Autoimmune Encephalomyelitis in Mice and Evaluation of the Disease-dependent Distribution of Immune Cells in Various Tissues.</a>&#160;J. Vis. Exp.&#160; (2016)
The video demonstrates the induction of experimental autoimmune encephalomyelitis in a mouse model by injecting a combination of myelin oligodendrocyte glycoproteins and inactivated Mycobacterium tuberculosis. The model enables researchers to study disease mechanisms and test therapeutic interventions.

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.
1. ...

Take an adjuvant emulsion containing myelin oligodendrocyte glycoproteins, or MOGs &#8212; a nerve myelin sheath protein, and inactivated Mycobacterium tuberculosis. Inject it subcutaneously into the mouse&#39;s back.
Next, intraperitoneally inject bacteria-derived exotoxins&#160; &#8212; disturbing the blood-brain barrier.
The subcutaneously injected Mycobacterium mimics local infection, attracting immune cells, including antigen-presenting cells or APCs.
APCs interact with MOGs, processing and presenting them on the major histocompatibility complexes, MHCs.
Activated APCs migrate to the lymph nodes, where T cells interact with APCs, becoming autoreactive T cells.
These T cells enter the bloodstream, cross the barrier, and infiltrate the central nervous system, CNS.
In the CNS, autoreactive T cells interact with MOG-expressing cells, promoting immune cell infiltration.
Infiltrated monocytes differentiate into macrophages, releasing inflammatory cytokines and reactive oxygen species, causing myelin-producing oligodendrocytes&#39; cell death.
B cells interact with autoreactive T cells, differentiating into plasma cells and secreting autoantibodies against MOG, leading to myelin sheath degradation.
The demyelination impairs nerve signal transmission, inducing autoimmune encephalomyelitis.

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Flow Cytometric Analysis of Lymphocyte Infiltration in Central Nervous System during Experimental Autoimmune Encephalomyelitis

Multiple sclerosis (MS) is an autoimmune disease of the central nervous system (CNS) caused by the combination of environmental factors and susceptible genetic background. Experimental autoimmune encephalomyelitis (EAE) is a typical disease model of MS widely used for investigating the pathogenesis in which T lymphocytes specific for myelin antigens initiate an inflammatory reaction in CNS. It is very important to assess how lymphocytes in the CNS regulate the development of disease. However, the approach for measuring the quantity and quality of infiltrated lymphocytes in the CNS is very limited due to the difficulties in isolating and detecting infiltrated lymphocytes from the brain. This manuscript presents a protocol for that is useful for the isolation, identification, and characterization of infiltrated lymphocytes from the CNS and will be helpful for our understanding of how lymphocytes are involved in the development of the CNS autoimmune disease.

This manuscript presents a protocol to induce active experimental autoimmune encephalomyelitis (EAE) in mice. A method for the isolation and characterization of the infiltrated lymphocytes in the central nervous system (CNS) is also presented to show how lymphocytes are involved in the development of CNS autoimmune disease.

实验自身免疫性脑脊髓炎中枢神经系统淋巴细胞渗透的流量细胞计量分析

Multiple sclerosis (MS) is an autoimmune disease of the central nervous system (CNS) caused by the combination of environmental factors and susceptible ...

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免疫与感染

副结核分枝杆菌在自身免疫性脑脊髓炎中的免疫原性增强

Adjuvant Activity of Mycobacterium paratuberculosis in Enhancing the Immunogenicity of Autoantigens During Experimental Autoimmune Encephalomyelitis

Experimental autoimmune encephalomyelitis (EAE) induced by myelin oligodendrocyte glycoprotein (MOG) requires immunization by a MOG peptide emulsified in complete Freund&#39;s adjuvant (CFA) containing inactivated Mycobacterium tuberculosis. The antigenic components of the mycobacterium activate dendritic cells to stimulate T-cells to produce cytokines that promote the Th1 response via toll-like receptors. Therefore, the amount and species of mycobacteria present during the antigenic challenge are directly related to the development of EAE. This methods paper presents an alternative protocol to induce EAE in C57BL/6 mice using a modified incomplete Freund&#39;s adjuvant containing the heat-killed Mycobacterium avium subspecies paratuberculosis strain K-10.
M. paratuberculosis, a member of the Mycobacterium avium complex, is the causative agent of Johne&#39;s disease in ruminants and has been identified as a risk factor for several human T-cell-mediated disorders, including multiple sclerosis. Overall, mice immunized with Mycobacterium paratuberculosis showed earlier onset and greater disease severity than mice immunized with CFA containing the strain of M. tuberculosis H37Ra at the same doses of 4 mg/mL. The antigenic determinants of Mycobacterium avium subspecies paratuberculosis (MAP)&#160;strain K-10 were able to induce a strong Th1 cellular response during the effector phase, characterized by significantly higher numbers of T-lymphocytes (CD4+ CD27+), dendritic cells (CD11c+ I-A/I-E+), and monocytes (CD11b+ CD115+) in the spleen compared to mice immunized with CFA. Furthermore, the proliferative T-cell response to the MOG peptide appeared to be highest in M. paratuberculosis-immunized mice. The use of an encephalitogen (e.g., MOG35-55) emulsified in an adjuvant containing M. paratuberculosis in the formulation may be an alternative and validated method to activate dendritic cells for priming myelin epitope-specific CD4+ T-cells during the induction phase of EAE.

作者聚焦: 副结核分枝杆菌在增强实验性自身免疫性脑脊髓炎期间自身抗原免疫原性中的辅助活性  

Here, we present an alternative protocol to actively induce experimental autoimmune encephalomyelitis in C57BL/6 mice, using the immunogenic epitope myelin oligodendrocyte glycoprotein (MOG)35-55 suspended in incomplete Freund's adjuvant containing the heat-killed Mycobacterium avium subspecies paratuberculosis.

副结核分枝杆菌增强实验性自身免疫性脑脊髓炎期间自身抗原免疫原的辅助活性

Experimental autoimmune encephalomyelitis (EAE) induced by myelin oligodendrocyte glycoprotein (MOG) requires immunization by a MOG peptide emulsified in ...

Induction and Diverse Assessment Indicators of Experimental Autoimmune Encephalomyelitis

Multiple sclerosis (MS) is a typical autoimmune disease of the central nervous system (CNS) characterized by inflammatory infiltration, demyelination, and axonal damage. Currently, there are no measures to cure MS completely, but multiple disease-modifying therapies (DMT) are available to control and mitigate disease progression. There are significant similarities between the CNS pathological features of experimental autoimmune encephalomyelitis (EAE) and MS patients. EAE has been widely used as a representative model to determine MS drugs' efficacy and explore the development of new therapies for MS disease. Active induction of EAE in mice has a stable and reproducible effect and is particularly suitable for studying the effects of drugs or genes on autoimmune neuroinflammation. The method of immunizing C57BL/6J mice with myelin oligodendrocyte glycoprotein (MOG35-55) and the daily assessment of disease symptoms using a clinical scoring system is mainly shared. Given the complex etiology of MS with diverse clinical manifestations, the existing clinical scoring system can't satisfy the assessment of disease treatment. To avoid the shortcomings of a single intervention, new indicators to assess EAE based on clinical manifestations of anxiety-like moods and osteoporosis in MS patients are created to provide a more comprehensive assessment of MS treatment.

The present protocol describes the induction of experimental autoimmune encephalomyelitis in a mouse model using myelin oligodendrocyte glycoprotein and monitoring the disease process using a clinical scoring system. Experimental autoimmune encephalomyelitis-related symptoms are analyzed using mouse femur micro-computed tomography analysis and open field test to assess the disease process comprehensively.

实验性自身免疫性脑脊髓炎的诱导及多样评价指标

Multiple sclerosis (MS) is a typical autoimmune disease of the central nervous system (CNS) characterized by inflammatory infiltration, demyelination, and ...

Induction of Experimental Autoimmune Encephalomyelitis in a Mouse Model

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Induction of Experimental Autoimmune Encephalomyelitis in a Mouse Model