Este trabajo fue apoyado por el NIH subvención DK080351 para PC.

Protocolo experimental El primer paso de este protocolo experimental es recoger un gran número de glándulas pituitarias de ratón. El segundo paso consiste en homogeneizar las glándulas para preparar un extracto de proteínas. El tercer paso es para emulsionar estas proteínas con una mezcla de hidrocarburos. Paso 1: Recolección y almacenamiento de ratón glándulas pituitarias La glándula pituitaria del ratón se encuentra en la base del cráneo en una depresión del hueso esfenoides llamada silla turca, rodeada lateralmente por los nervios trigémino y anteriormente por el quiasma óptico. La glándula está compuesta de un lóbulo anterior más grande y más pequeño lóbulo posterior y medio, y pesa un promedio de 1,9 mg. La glándula pituitaria son recogidos de los ratones de cepas mixta, sexos, edades, y destinado a ser sacrificado por el centro de atención animal. Para aislar la glándula pituitaria, el ratón es la eutanasia, la piel sobre el cráneo disecado, y el hueso del cráneo a cielo abierto con unas tijeras. El cerebro se levanta para exponer la glándula pituitaria, la cual es entonces aislado de los alrededores de la duramadre, recogió del hueso esfenoides y se almacena en un tubo de plástico con hielo seco. En general, se utilizan 300 glándulas pituitaria del ratón para la preparación de una proteína, un número que puede ser recogido por una persona en 9 horas. Una vez que la colección se completa, el tubo que contiene las glándulas pituitarias se almacenan a -80 ° C hasta que esté listo para el siguiente paso. Paso 2. La extracción de las proteínas del ratón pituitaria <ol><li> Añadir el tampón de homogeneización a un tubo Falcon de 15 ml (catálogo no 352.059), mantenidos en un baño de hielo, con 250 l de tampón por cada 100 glándulas pituitarias recogidos. Utilice un tampón de fuerza iónica similar a la que es fisiológicamente el interior del citoplasma de la célula (0,2 M) y de pH fisiológico (7,4). Usamos tampón fosfato salino (0,161 M, pH 7,4), complementado con un cóctel de inhibidores de la proteasa (Sigma-Aldrich, el catálogo no P8340-1 ml). </li><li> Añadir la glándula pituitaria en el búfer antes de la homogeneización, y luego colocar el generador (5 mm de diámetro) de la homogeneizador Polytron en la parte inferior del tubo Falcon. Homogeneizar a máximas rpm durante 30 segundos, moviendo suavemente el tubo Falcon arriba y hacia abajo mientras lo mantiene en el baño de hielo. Luego, el resto del homogeneizado en hielo durante 1 min. Repita la homogeneización y refrigeración pasos dos veces más. Esta homogeneización suave debe romper las células y liberar las proteínas citosólicas sin interrumpir los núcleos. </li><li> Para eliminar los núcleos, el tejido roto y materiales insolubles (por ejemplo, tejido conectivo y los complejos de proteínas desnaturalizadas), se centrifuga el homogeneizado a baja velocidad (1000 g) durante 10 min a 4 ° C. </li><li> Transferir el sobrenadante (que ahora se llama post-nuclear sobrenadante, PNS) a un nuevo tubo Falcon, teniendo cuidado de dejar la bolita sin tocar, y almacenar el SNP en el hielo. </li><li> Resuspender el precipitado en el volumen original de tampón de homogeneización (250 l de tampón por cada 100 glándulas pituitarias recogidos), homogeneizar el anterior, se centrifuga como antes, y luego combinar el SNP segundo con el primero. </li><li> Alícuota de los dos combinados PNS muestras en tubos de microcentrífuga y se almacena a -80 ° C hasta que esté listo para el siguiente paso. Esta preparación de proteínas puede ser almacenado a -80 ° C, pero poco a poco pierde su potencia, lo que se recomienda para utilizar antes de 6 meses a partir de la preparación. Use una pequeña alícuota para determinar el rendimiento de proteína y la concentración por el ensayo BCA (Pierce catálogo no. 23.225), y la calidad de las proteínas por electroforesis en gel. En general, se obtienen alrededor de 60 mg de proteína de 300 hipófisis de ratón en 1,5 ml de tampón de homogeneización, a una concentración de alrededor de 40 mg / ml. </li></ol> Paso 3. Preparación de la emulsión para la inmunización <ol><li> Descongele rápidamente las alícuotas congeladas proteína necesaria para el experimento y ajustar la concentración de proteínas y 20 mg / ml. Este extracto de proteínas se emulsiona 1:1 con adyuvante completo de Freund (CFA, Sigma F-5881) que contiene 5 mg / mL de calor mató a Mycobacterium tuberculosis (Becton Dickinson, 231 141). Cada ratón recibirá 1 mg de extracto de pituitaria en 100 L de la emulsión. </li><li> El ejemplo anterior es para inmunizar a 10 ratones, en la que cada ratón inyectado con 100 L de la emulsión que contiene 1 mg de proteínas de la hipófisis. Permitir que dos ratones más por la pérdida de material durante la preparación, a fin de utilizar 12 mg de proteínas de la hipófisis. Aspire 600 l de los 20 mg / ml de extracto de proteína de la pituitaria anterior describe con una jeringa de 2,5 ml Hamilton gas. Resuspender el Comité de Libertad Sindical por agitación, y aspiran 600 l de CFA en otra jeringa de 2,5 ml Hamilton. Adjuntar y asegurar una de calibre 22 micro-emulsión aguja en la jeringa llena de CFA. Empuje lentamente el émbolo de la jeringa CFA de tal manera que la aguja está lleno de CFA, a continuación, colocar y fijar el otro extremo de lala aguja de la jeringa llena con extracto de pituitaria. </li><li> Avanzar lentamente el émbolo de la jeringa CFA para impulsar el componente de aceite en el extracto acuoso pituitaria, de tal manera que sólo una pequeña cantidad se mezcla. A continuación, empuje el émbolo de la jeringa extracto de pituitaria en la jeringa CFA. Repita esta acción aumentando gradualmente la cantidad de material mezclado. Continúe hasta que todo el contenido de las dos jeringas se mezclan. Usted obtendrá una solución blanquecina y uniformemente distribuida que representa la emulsión de agua en aceite. </li><li> Una vez que la emulsión se forma, repita el movimiento hacia adelante y hacia atrás por lo menos 500 veces para asegurarse de que la emulsión se mezcla bien. Después de que el ciclo de mezclado final, la 1:01 de agua en emulsión de aceite está listo para usar y contiene una concentración de proteína pituitaria de 10 mg / ml. La emulsión se inyectó el mismo día por vía subcutánea en ratones anestesiados SJL, como se detalla en el artículo JoVe compañero. </li></ol>

<ol>
	<li>Caturegli, P., Newschaffer, C., Olivi, A., Pomper, M. G., Burger, P. C., Rose, N. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Autoimmune Hypophysitis Endocr Rev. 26, 599-614 (2005).</li><li>Blansfield, J. A., Beck, K. E., Tran, K., Yang, J. C., Hughes, M. S., Kammula, U. S., Royal, R. E., Topalian, S. L., Haworth, L. R., Levy, C., Rosenberg, S. A., Sherry, R. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cytotoxic+T-lymphocyte-associated+antigen-4+blockage+can+induce+autoimmune+hypophysitis+in+patients+with+metastatic+melanoma+and+renal+cancer.">Cytotoxic T-lymphocyte-associated antigen-4 blockage can induce autoimmune hypophysitis in patients with metastatic melanoma and renal cancer.</a> J Immunother. 28, 593-598 (2005).</li><li>Tzou, S. C., Lupi, I., Landek, M., Gutenberg, A., Tzou, Y. M., Kimura, H., Pinna, G., Rose, N. R., Caturegli, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Autoimmune+Hypophysitis+of+SJL+mice:+Clinical+Insights+from+a+New+Animal+Model.">Autoimmune Hypophysitis of SJL mice: Clinical Insights from a New Animal Model.</a> Endocrinology. 149, 3461-3469 (2008).</li><li>Goudie, R. B., Pinkerton, P. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Anterior+hypophysitis+and+Hashimoto's+disease+in+a+woman.">Anterior hypophysitis and Hashimoto's disease in a woman.</a> J Pathol Bacteriol. 83, 584-585 (1962).</li><li>Caturegli, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Autoimmune+hypophysitis:+an+underestimated+disease+in+search+of+its+autoantigen(s).">Autoimmune hypophysitis: an underestimated disease in search of its autoantigen(s).</a> J Clin Endocrinol Metab. 92, 2038-2040 (2007).</li></ol>

A pesar de que el primer paciente con hipofisitis autoinmune se informó en 1962 4, los autoantígenos patógenos hipófisis aún no se han identificado 5. Los modelos animales pueden ser muy útiles para identificar a estos autoantígenos, ayudar al descubrimiento de biomarcadores que son traducibles al cuidado del paciente. Este artículo ha demostrado un procedimiento sencillo para la preparación de proteínas de la hipófisis en una forma que es adecuada para la inducción de hipofisitis autoi...

<table><tbody><tr><td>15-ml Falcon tubes</td><td></td><td></td><td></td></tr><tr><td>homogenization buffer</td><td></td><td></td><td></td></tr><tr><td>phosphate buffered saline</td><td></td><td></td><td></td></tr><tr><td>protease inhibitor cocktail</td><td></td><td></td><td></td></tr><tr><td>Polytron homogenizer</td><td></td><td></td><td></td></tr><tr><td>BCA assay</td><td>Pierce, Thermo Scientific</td><td>23225</td><td></td></tr><tr><td>Complete Freund&amp;rsquo;s Adjuvant (CFA)</td><td>Sigma-Aldrich</td><td>F-5881</td><td></td></tr><tr><td>Mycobacterium tuberculosis</td><td>BD Biosciences</td><td>231141</td><td></td></tr><tr><td>2.5 mL Hamilton gastight syringe</td><td></td><td></td><td></td></tr><tr><td>22-gauge micro-emulsifying needle</td><td></td><td></td><td></td></tr></tbody></table>

preparation of mouse pituitary immunogen for the induction of experimental autoimmune hypophysitis

Hipofisitis autoinmune es una inflamación crónica de la glándula pituitaria causado o acompañado por una autoinmunidad. Tradicionalmente se ha considerado una enfermedad rara, pero la información se ha incrementado notablemente en los últimos años. Hipofisitis, de hecho, no es raro desarrolla como un &quot;efecto secundario&quot; en pacientes con cáncer tratados con anticuerpos que bloquean los receptores inhibitorios expresa en los linfocitos T, tales como CTLA-4 2 y los receptores de PD-1. Hipofisitis autoinmune puede ser inducida experimentalmente mediante la inyección de ratones con las proteínas de la hipófisis se mezcla con un adyuvante 3. En este artículo de vídeo que muestra cómo extraer las proteínas de ratón glándulas pituitaria y la forma de prepararlos de forma adecuada para la inducción de hipofisitis autoinmune en ratones SJL.

Watch this Scientific Journal Video about Preparation of Mouse Pituitary Immunogen for the Induction of Experimental Autoimmune Hypophysitis at JoVE.com

Preparation of Mouse Pituitary Immunogen for the Induction of Experimental Autoimmune Hypophysitis

Hipofisitis autoinmune se puede reproducir en ratones mediante la inyección de un extracto de proteínas de ratón pituitaria.

Preparación de inmunógeno ratón hipófisis para la inducción de la hipofisitis autoinmune experimental

Autoimmune hypophysitis is a chronic inflammation of the pituitary gland caused or accompanied by autoimmunity1.  It has traditionally been considered a ...

preparation-mouse-pituitary-immunogen-for-induction-experimental

Universidad de Cartagena UDEC

Research

JoVE Journal

Immunology and Infection

14.5K vistas.  The Johns Hopkins University. Hipofisitis autoinmune se puede reproducir en ratones mediante la inyección de un extracto de proteínas de ratón pituitaria.

Video: Preparation of Mouse Pituitary Immunogen for the Induction of Experimental Autoimmune Hypophysitis

Induction of Experimental Autoimmune Hypophysitis in SJL Mice

Autoimmune hypophysitis can be reproduced experimentally by the injection of pituitary proteins mixed with an adjuvant into susceptible mice1. Mouse models allow us to study how diseases unfold, often providing a good replica of the same processes occurring in humans. For some autoimmune diseases, like type 1A diabetes, there are models (the NOD mouse) that spontaneously develop a disease similar to the human counterpart. For many other autoimmune diseases, however, the model needs to be induced experimentally. A common approach in this regard is to inject the mouse with a dominant antigen derived from the organ being studied. For example, investigators interested in autoimmune thyroiditis inject mice with thyroglobulin2, and those interested in myasthenia gravis inject them with the acetylcholine receptor3. If the autoantigen for a particular autoimmune disease is not known, investigators inject a crude protein extract from the organ targeted by the autoimmune reaction. For autoimmune hypophysitis, the pathogenic autoantigen(s) remain to be identified4, and thus a crude pituitary protein preparation is used. In this video article we demonstrate how to induce experimental autoimmune hypophysitis in SJL mice.

This video shows how to induce autoimmune hypophysitis in SJL mice and how to assess its severity by histopathology.

La inducción de la hipofisitis autoinmune experimental en ratones SJL

Autoimmune hypophysitis can be reproduced experimentally by the injection of pituitary proteins mixed with an adjuvant into susceptible mice1. Mouse ...

Investigación

Inmunología e Infección

Overcoming Unresponsiveness in Experimental Autoimmune Encephalomyelitis (EAE) Resistant Mouse Strains by Adoptive Transfer and Antigenic Challenge

Experimental autoimmune encephalomyelitis (EAE) is an inflammatory disease of the central nervous system (CNS) and has been used as an animal model for study of the human demyelinating disease, multiple sclerosis (MS). EAE is characterized by pathologic infiltration of mononuclear cells into the CNS and by clinical manifestation of paralytic disease. Similar to MS, EAE is also under genetic control in that certain mouse strains are susceptible to disease induction while others are resistant. Typically, C57BL/6 (H-2b) mice immunized with myelin basic protein (MBP) fail to develop paralytic signs. This unresponsiveness is certainly not due to defects in antigen processing or antigen presentation of MBP, as an experimental protocol described here had been used to induce severe EAE in C57BL/6 mice as well as other reputed resistant mouse strains. In addition, encephalitogenic T cell clones from C57BL/6 and Balb/c mice reactive to MBP had been successfully isolated and propagated.
The experimental protocol involves using a cellular adoptive transfer system in which MBP-primed (200 &mu;g/mouse) C57BL/6 donor lymph node cells are isolated and cultured for five days with the antigen to expand the pool of MBP-specific T cells. At the end of the culture period, 50 million viable cells are transferred into naive syngeneic recipients through the tail vein. Recipient mice so treated normally do not develop EAE, thus reaffirming their resistant status, and they can remain normal indefinitely. Ten days post cell transfer, recipient mice are challenged with complete Freund adjuvant (CFA)-emulsified MBP in four sites in the flanks. Severe EAE starts to develop in these mice ten to fourteen days after challenge. Results showed that the induction of disease was antigenic specific as challenge with irrelevant antigens did not induce clinical signs of disease. Significantly, a titration of the antigen dose used to challenge the recipient mice showed that it could be as low as 5 &mu;g/mouse. In addition, a kinetic study of the timing of antigenic challenge showed that challenge to induce disease was effective as early as 5 days post antigenic challenge and as long as over 445 days post antigenic challenge. These data strongly point toward the involvement of a "long-lived" T cell population in maintaining unresponsiveness. The involvement of regulatory T cells (Tregs) in this system is not defined.

Overcoming Unresponsiveness in Experimental Autoimmune Encephalomyelitis EAE Resistant Mouse Strains by Adoptive Transfer and Antigenic Challenge

Certain mouse strains are able to resist induction of experimental autoimmune encephalomyelitis (EAE) with myelin basic protein. Described here is a simple immunization protocol that reverses the unresponsiveness and induces paralytic disease in several typical EAE resistant mouse stains.

Superar la falta de respuesta en encefalomielitis autoinmune experimental (EAE) cepas de ratones resistentes a través de transferencia adoptiva y la estimulación antigénica

Experimental autoimmune encephalomyelitis (EAE) is an inflammatory disease of the central nervous system (CNS) and has been used as an animal model for ...

Systemic Injection of Neural Stem/Progenitor Cells in Mice with Chronic EAE

Neural stem/precursor cells (NPCs) are a promising stem cell source for transplantation approaches aiming at brain repair or restoration in regenerative neurology. This directive has arisen from the extensive evidence that brain repair is achieved after focal or systemic NPC transplantation in several preclinical models of neurological diseases.
These experimental data have identified the cell delivery route as one of the main hurdles of restorative stem cell therapies for brain diseases that requires urgent assessment. Intraparenchymal stem cell grafting represents a logical approach to those pathologies characterized by isolated and accessible brain lesions such as spinal cord injuries and Parkinson's disease. Unfortunately, this principle is poorly applicable to conditions characterized by a multifocal, inflammatory and disseminated (both in time and space) nature, including multiple sclerosis (MS). As such, brain targeting by systemic NPC delivery has become a low invasive and therapeutically efficacious protocol to deliver cells to the brain and spinal cord of rodents and nonhuman primates affected by experimental chronic inflammatory damage of the central nervous system (CNS).
This alternative method of cell delivery relies on the NPC pathotropism, specifically their innate capacity to (i) sense the environment via functional cell adhesion molecules and inflammatory cytokine and chemokine receptors; (ii) cross the leaking anatomical barriers after intravenous (i.v.) or intracerebroventricular (i.c.v.) injection; (iii) accumulate at the level of multiple perivascular site(s) of inflammatory brain and spinal cord damage; and (i.v.) exert remarkable tissue trophic and immune regulatory effects onto different host target cells in vivo.
Here we describe the methods that we have developed for the i.v. and i.c.v. delivery of syngeneic NPCs in mice with experimental autoimmune encephalomyelitis (EAE), as model of chronic CNS inflammatory demyelination, and envisage the systemic stem cell delivery as a valuable technique for the selective targeting of the inflamed brain in regenerative neurology.

The transplantation of neural stem/progenitor cells (NPCs) holds great promises in regenerative neurology. The systemic delivery of NPCs has turned into effective, low invasive, and therapeutically very efficacious protocol to deliver stem cells in the brain and spinal cord of rodents and nonhuman primates affected by experimental chronic inflammatory damage of the central nervous system.

Inyección sistémica de Neuronales células madre / progenitoras en ratones con EAE crónica

Neural stem/precursor cells (NPCs) are a promising stem cell source for transplantation approaches aiming at brain repair or restoration in regenerative ...

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.

Un enfoque Simple para inducir Neuritis Autoinmune Experimental en ratones C57BL/6 para las evaluaciones funcionales y Neuropathological

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

Neurociencias

Experimental Autoimmune Uveitis: An Intraocular Inflammatory Mouse Model

Experimental Autoimmune Uveitis (EAU) is driven by immune cells responding to self-antigens. Many features of this non-infectious, intraocular inflammatory disease model recapitulate the clinical phenotype of posterior uveitis affecting humans. EAU has been used reliably to study the efficacy of novel inflammatory therapeutics, their mode of action and to further investigate the mechanisms that underpin disease progression of intraocular disorders. Here, we provide a detailed protocol on EAU induction in the C57BL/6J mouse - the most widely used model organism with susceptibility to this disease. Clinical assessment of disease severity and progression will be demonstrated using fundoscopy, histological examination and fluorescein angiography. The induction procedure involves subcutaneous&#160;injection of an emulsion containing a peptide (IRBP1-20) from the ocular protein interphotoreceptor retinoid binding protein (also known as retinol binding protein 3), Complete Freund&#39;s Adjuvant (CFA) and supplemented with killed Mycobacterium tuberculosis. Injection of this viscous emulsion on the back of the neck is followed by a single intraperitoneal injection of Bordetella pertussis toxin. At the onset of symptoms (day 12-14) and under general anesthesia, fundoscopic images are taken to assess disease progression through clinical examination. These data can be directly compared with those at later timepoints and peak disease (day 20-22) with differences analyzed. At the same time, this protocol allows the investigator to assess potential differences in vessel permeability and damage using fluorescein angiography. EAU can be induced in other mouse strains - both wildtype or genetically modified - and combined with novel therapies offering flexibility for studying drug efficacy and/or disease mechanisms.

In this report we present a protocol that allows the investigator to generate a mouse model of intraocular uveitis. More commonly referred to as experimental autoimmune uveitis (EAU), this established model captures many aspects of human disease. Herein, we will describe how to induce and monitor disease progression using several readouts.

Uveítis autoinmune experimental: un modelo de ratón inflamatorio intraocular

Experimental Autoimmune Uveitis (EAU) is driven by immune cells responding to self-antigens. Many features of this non-infectious, intraocular ...

A Rapid, Simple, and Standardized Homogenization Method to Prepare Antigen/Adjuvant Emulsions for Inducing Experimental Autoimmune Encephalomyelitis

Experimental autoimmune encephalomyelitis (EAE) shares similar immunological and clinical features with multiple sclerosis (MS), and is therefore widely used as a model to identify new drug targets for better patient treatment. MS is characterized by several different disease courses: relapsing-remitting MS (RRMS), primary progressive MS (PPMS), secondary progressive MS (SPMS), and a rare progressive-relapsing form of MS (PRMS). Although animal models do not accurately mimic all of these contrasting human disease phenotypes, there are EAE models that reflect some of the different clinical manifestations of MS. For example, myelin oligodendrocyte glycoprotein (MOG)-induced EAE in C57BL/6J mice mimics human PPMS, while myelin proteolipid protein (PLP)-induced EAE in SJL/J mice resembles RRMS. Other autoantigens, such as myelin basic protein (MBP), and a number of different mouse strains are also used to study EAE. To induce disease in these autoantigen-immunization EAE models, a water-in-oil emulsion is prepared and injected subcutaneously. The majority of EAE models also require an injection of pertussis toxin for the disease to develop. For consistent and reproducible EAE induction, a detailed protocol to prepare the reagents to produce antigen/adjuvant emulsions is necessary. The method described here takes advantage of a standardized method to generate water-in-oil emulsions. It is simple and fast and uses a shaking homogenizer instead of syringes to prepare quality-controlled emulsions.

To induce experimental autoimmune encephalomyelitis, an animal model of multiple sclerosis, mice are immunized with a water-in-oil emulsion containing an autoantigen and complete Freund's adjuvant. While several protocols exist for the preparation of these emulsions, a rapid, simple, and standardized homogenization protocol for emulsion preparation is presented here.

Un método de homogeneización rápido, simple y estandarizado para preparar emulsiones de antígeno/adyuvante para inducir encefalomielitis autoinmune experimental

Experimental autoimmune encephalomyelitis (EAE) shares similar immunological and clinical features with multiple sclerosis (MS), and is therefore widely ...

Generation of a Mouse Spontaneous Autoimmune Thyroiditis Model

In recent years, Hashimoto's thyroiditis (HT) has become the most common autoimmune thyroid disease. It is characterized by lymphocyte infiltration and the detection of specific serum autoantibodies. Although the potential mechanism is still not clear, the risk of Hashimoto's thyroiditis is related to genetic and environmental factors. At present, there are several types of models of autoimmune thyroiditis, including experimental autoimmune thyroiditis (EAT) and spontaneous autoimmune thyroiditis (SAT).
EAT in mice is a common model for HT, which is immunized with lipopolysaccharide (LPS) combined with thyroglobulin (Tg) or supplemented with complete Freund's adjuvant (CFA). The EAT mouse model is widely established in many types of mice. However, the disease progression is more likely associated with the Tg antibody response, which may vary in different experiments.
SAT is also widely used in the study of HT in the NOD.H-2h4 mouse. The NOD.H2h4 mouse is a new strain obtained from the cross of the nonobese diabetic (NOD) mouse with the B10.A(4R), which is significantly induced for HT with or without feeding iodine. During the induction, the NOD.H-2h4 mouse has a high level of TgAb accompanied by lymphocyte infiltration in the thyroid follicular tissue. However, for this type of mouse model, there are few studies to comprehensively evaluate the pathological process during the induction of iodine.
A SAT mouse model for HT research is established in this study, and the pathologic changing process is evaluated after a long period of iodine induction. Through this model, researchers can better understand the pathological development of HT and screen new treatment methods for HT.

Several types of animal models of Hashimoto's thyroiditis have been established, as has spontaneous autoimmune thyroiditis in the NOD mouse. H-2h4 mice are a simple and reliable model for HT induction. This article describes this approach and evaluates the pathological process for a better understanding of the SAT murine model.

Generación de un modelo de tiroiditis autoinmune espontánea en ratones

In recent years, Hashimoto's thyroiditis (HT) has become the most common autoimmune thyroid disease. It is characterized by lymphocyte infiltration and ...

Inducción y evaluación de la encefalomielitis autoinmune experimental en ratones

Modeling Multiple Sclerosis in the Two Sexes: MOG35-55-Induced Experimental Autoimmune Encephalomyelitis

Multiple Sclerosis (MS) is a chronic autoimmune inflammatory disease affecting the central nervous system (CNS). It is characterized by different prevalence in the sexes, affecting more women than men, and different outcomes, showing more aggressive forms in men than in women. Furthermore, MS is highly heterogeneous in terms of clinical aspects, radiological, and pathological features. Thus, it is necessary to take advantage of experimental animal models that allow the investigation of as many aspects of the pathology as possible. Experimental autoimmune encephalomyelitis (EAE) represents one of the most used models of MS in mice, modeling different disease features, from the activation of the immune system to CNS damage. Here we describe a protocol for the induction of EAE in both male and female C57BL/6J mice using myelin oligodendrocyte glycoprotein peptide 35-55 (MOG35-55) immunization, which leads to the development of a chronic form of the disease. We also report the evaluation of the daily clinical score and motor performance of these mice for 28 days post immunization (28 dpi). Lastly, we illustrate some basic histological analysis at the CNS level, focusing on the spinal cord as the primary site of disease-induced damage.

Autor destacado: Creación de un modelo experimental versátil de encefalomielitis autoinmune relevante para ratones machos y hembras 

Experimental autoimmune encephalomyelitis is one of the most widely used murine models of multiple sclerosis. In the current protocol, C57BL/6J mice of both sexes are immunized with myelin oligodendrocyte glycoprotein peptide, resulting mainly in ascending paresis of the tail and limbs. Here we discuss the protocol of EAE induction and evaluation.

Preparación de inmunógeno ratón hipófisis para la inducción de la hipofisitis autoinmune experimental

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