eoe sub articles

EoE Sub Articles

1. EAE Induction in SJL/J Mice
<ol>
	<li>Mice
	<ol>
		<li>Use 11-week-old female SJL/J mice and allow them to habituate to the experimental room for about 7 days. Use n = 10 mice per group.</li>
		<li>To assess medication effects, continuously administer the drug and placebo for the control group via drinking water or food pellets starting 3 or 5 days after immunization (n = 10 per group). During the peak of the disease, administer medication or placebo with milk or 3% sugar water-soaked cornflakes.</li>
	</ol>
	</li>
	<li>Immunization material
	<ol>
		<li>Use an EAE induction kit consisting of antigen (peptide of proteolipid protein, PLP139-151, 1 mg/mL emulsion) in an emulsion with complete Freund&#39;s adjuvant (CFA, heat-killed Mycobacterium tuberculosis H37 Ra) and 2 vials (5 &#956;g each) of lyophilized pertussis toxin (PTX).</li>
		<li>Dissolve the PTX (2 &#956;g/mL) in 1x phosphate-buffered saline (PBS, i.e., add 1.5 mL of PBS to each PTX tube, mix well, remove with the same pipet tip, and add to 1 mL of PBS in a 50-mL tube); mix well.</li>
	</ol>
	</li>
	<li>Immunization
	<ol>
		<li>Inject PLP/CFA subcutaneously in 2 portions, each 100 &#956;L, both at the base of the tail.</li>
		<li>Inject 100 &#956;L of PTX intraperitoneally (i.p.) twice per mouse, the first 1 - 2 h after immunization and the second at 24 h.</li>
		<li>For the control mice, inject CFA without PLP (2 portions of 100 &#956;L) plus PTX without PLP.</li>
	</ol>
	</li>
</ol>

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>PLP139-151/CFA emulsion</td>
			<td>Hooke Labs, St Lawrence, MA</td>
			<td>EK-0123</td>
			<td>EAE induction kit</td>
		</tr>
		<tr>
			<td>Pertussis Toxin</td>
			<td>Hooke Labs, St Lawrence, MA</td>
			<td>EK-0123</td>
			<td>EAE induction kit</td>
		</tr>
		<tr>
			<td>IVIS Lumina Spectrum</td>
			<td>Perkin Elmer, Inc., Waltham, USA</td>
			<td></td>
			<td>Bioluminescence and Infrared Imaging System</td>
		</tr>
		<tr>
			<td>LivingImage 4.5 software</td>
			<td>Perkin Elmer, Inc., Waltham, USA</td>
			<td>CLS136334</td>
			<td>IVIS analysis software</td>
		</tr>
		<tr>
			<td>Isoflurane</td>
			<td>Abbott Labs, Illinois, USA</td>
			<td>26675-46-7</td>
			<td>Anaesthetic</td>
		</tr>
	</tbody>
</table>

induction of experimental autoimmune encephalomyelitis in a mouse model

Source: Schmitz, K., et al. <a href="https://appjove.unicartagenaproxy.elogim.com/v/55321">Bioluminescence and Near-infrared Imaging of Optic Neuritis and Brain Inflammation in the EAE Model of Multiple Sclerosis in Mice.</a> J. Vis. Exp (2017).
In this video, a mouse model of multiple sclerosis is established using experimental autoimmune encephalomyelitis (EAE). The mouse is injected with an emulsion containing a neuronal peptide and an adjuvant, triggering an immune response that activates autoreactive T cells. This is followed by an injection of pertussis toxin (PTX) to increase blood-brain barrier permeability, allowing the autoreactive T-cells to infiltrate the brain. This infiltration initiates a signaling cascade that ultimately results in myelin damage, mimicking the demyelinating condition of multiple sclerosis.

Induction of Experimental Autoimmune Encephalomyelitis in a Mouse Model

1. EAE Induction in SJL/J Mice

	Mice
	
		Use 11-week-old female SJL/J mice and allow them to habituate to the experimental room for about 7 days. Use n = ...

Begin by subcutaneously injecting an emulsion containing a neuronal peptide and a heat-killed bacterial adjuvant into an anesthetized mouse.
In the subcutaneous tissue, the adjuvant attracts antigen-presenting cells, or APCs, which process and present the neuronal peptides on major histocompatibility complexes.
The activated APCs reach the lymph nodes and interact with na&#239;ve T cells, inducing their differentiation into autoreactive T cells.
These T cells circulate and reach the blood-brain barrier or BBB.
Next, intraperitoneally inject bacteria-derived exotoxins.
The exotoxins reach the BBB, increasing its permeability and allowing T cell infiltration into the central nervous system (CNS).
In the CNS, T cells interact with resident APCs presenting the neuronal peptide, promoting cytokine release that induces further immune cell infiltration.
The immune cells release pro-inflammatory cytokines, causing myelin damage.
Additionally, B-cells interact with T cells and secrete autoantibodies that further damage myelin, establishing experimental autoimmune encephalomyelitis (EAE).

induction-experimental-autoimmune-encephalomyelitis-mouse

Universidad de Cartagena UDEC

Research

JoVE Encyclopedia of Experiments

Neuroscience

Neuropathology

Neuromuscular Disorders

37 Views. Source: Schmitz, K., et al. Bioluminescence and Near-infrared Imaging of Optic Neuritis and Brain Inflammation in the EAE Model of Multiple Sclerosis in Mice. J. Vis. Exp (2017). In this video, a mouse model of multiple sclerosis is established using experimental autoimmune encephalomyelitis (EAE). The mouse is injected with an emulsion containing a neuronal peptide and an adjuvant, triggering an immune response that activates autoreactive T cells. This is followed by an injection of pertussis ...

Video: Induction of Experimental Autoimmune Encephalomyelitis in a Mouse Model - Concept

Bioluminescence Imaging to Detect Late Stage Infection of African Trypanosomiasis

Human African trypanosomiasis (HAT) is a multi-stage disease that manifests in two stages; an early blood stage and a late stage when the parasite invades the central nervous system (CNS). In vivo study of the late stage has been limited as traditional methodologies require the removal of the brain to determine the presence of the parasites.
Bioluminescence imaging is a non-invasive, highly sensitive form of optical imaging that enables the visualization of a luciferase-transfected pathogen in real-time. By using a transfected trypanosome strain that has the ability to produce late stage disease in mice we are able to study the kinetics of a CNS infection in a single animal throughout the course of infection, as well as observe the movement and dissemination of a systemic infection.
Here we describe a robust protocol to study CNS infections using a bioluminescence model of African trypanosomiasis, providing real time non-invasive observations which can be further analyzed with optional downstream approaches.

This manuscript describes the use of a bioluminescent strain of African trypanosomes to enable the tracking of late stage infection and demonstrates how in vivo live imaging can be used to visualize infections within the central nervous system in real-time.

Human African trypanosomiasis (HAT) is a multi-stage disease that manifests in two stages; an early blood stage and a late stage when the parasite invades ...

JoVE Journal

Immunology and Infection

Bioluminescence Imaging of an Immunocompetent Animal Model for Glioblastoma

In contrast to commonly reported human glioma xenograft animal models, GL261 murine glioma xenografts recapitulate nearly all relevant clinical and histopathologic features of the human disease. When GL261 cells are implanted intracranially in syngeneic C57BL/6 mice, the model has the added advantage of maintaining an intact immune microenvironment. Stable expression of luciferase in GL261 cells allows non-invasive cost effective bioluminescence monitoring of intracranial tumor growth. We have recently demonstrated that luciferase expression in GL261 cells does not affect the tumor growth properties, tumor cell immunomodulatory cytokine expression, infiltration of immune cells into the tumor, or overall survival of animals bearing the intracranial tumor. Therefore, it appears that the GL261 luciferase glioma model can be useful in the study of novel chemotherapeutic and immunotherapeutic modalities. Here we report the technique for generating stable luciferase expression in GL261 cells and how to study the in vitro and in vivo growth of the tumor cells by bioluminescence imaging.

GL261 glioma cells provide a useful immunocompetent animal model of glioblastoma. The goals of this protocol are to demonstrate proper techniques for monitoring intracranial tumor growth using in vivo bioluminescence imaging, and to verify the utility of luciferase-modified GL261 cells for studying tumor immunology and immunotherapeutic approaches for treating glioblastoma.

In contrast to commonly reported human glioma xenograft animal models, GL261 murine glioma xenografts recapitulate nearly all relevant clinical and ...

Medicine

A Simple Approach to Induce Experimental Autoimmune Neuritis in C57BL/6 Mice for Functional and Neuropathological Assessments

Experimental autoimmune neuritis (EAN) is a well-appreciated experimental model of autoimmune peripheral demyelinating diseases. EAN disease is induced by immunizing mice with neurogenic peptides to direct an inflammatory attack toward components of the peripheral nervous system (PNS). Recent advances have enabled the induction of EAN in the relatively resistant C57BL/6 mouse line using myelin protein zero (P0)106-125 or P0180-199 peptides delivered in adjuvant combined with the injection of pertussis toxin. The ability to induce EAN in the C57BL/6 strain allows for the use of the numerous genetic tools that exist on this mouse background, and thus allows the sophisticated study of disease pathogenesis and interrogation of the mechanistic action of novel therapeutics in combination with transgenic approaches. In this study, we demonstrate a simple approach to successfully induce EAN using the P0180-199 peptide in C57BL/6 mice. We also outline a protocol for the assessment of functional deficits that occur in this model, accompanied by an array of neuropathological features. Thus, this model is a powerful experimental model to study the pathogenesis of human peripheral demyelinating neuropathies, and to determine the efficacy of potential therapies that aim to promote myelin repair and protect against nerve damage in autoimmune neuritis.

This report outlines a simple approach to successfully induce experimental autoimmune neuritis (EAN) using the myelin protein zero (P0)180-199 peptide in combination with Freund&#39;s complete adjuvant and pertussis toxin. We present a sophisticated paradigm capable of accurately assessing the extent of functional deficits and neuropathology that occur in this EAN.

Experimental autoimmune neuritis (EAN) is a well-appreciated experimental model of autoimmune peripheral demyelinating diseases. EAN disease is induced by ...

Induction of Experimental Autoimmune Encephalomyelitis in a Mouse Model

Transcript

Induction of Experimental Autoimmune Encephalomyelitis in a Mouse Model

Bioluminescence Imaging to Detect Late Stage Infection of African Trypanosomiasis

Bioluminescence Imaging of an Immunocompetent Animal Model for Glioblastoma

A Simple Approach to Induce Experimental Autoimmune Neuritis in C57BL/6 Mice for Functional and Neuropathological Assessments