معتمدة في جزء من المنح المقدمة من المعاهد الوطنية للصحة (R01 NS37253 وNS055167 N01) والجمعية الوطنية لمرض التصلب العصبي المتعدد (RG-3288 A6).

<ol>
	<li>Swanborg, R. H., Sabato, G. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Experimental+allergic+encephalomyelitis.">Experimental allergic encephalomyelitis.</a> Methods in Enzymology. 162, 413-421 (1988).</li><li>Shaw, M. K., Kim, C., Hao, H. W., Chen, F., Tse, H. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Induction+of+myelin+basic+protein-specific+experimental+autoimmune+encephalomyelitis+in+C57BL/6+mice:+Mapping+of+T+cell+epitopes+and+T+cell+Vb+gene+segment+usage.">Induction of myelin basic protein-specific experimental autoimmune encephalomyelitis in C57BL/6 mice: Mapping of T cell epitopes and T cell Vb gene segment usage.</a> J. Neuroscience Research. 45, 690-699 (1996).</li><li>McCarron, R., McFarlin, D. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Adoptively+transferred+experimental+autoimmune+encephalomyelitis+in+SJL/J,+PL/J,+and+(SJL/J+x+PL/J)F1+mice.+Influence+of+I-A+haplotypes+on+encephalitogenic+epitope+of+myelin+basic+protein.">Adoptively transferred experimental autoimmune encephalomyelitis in SJL/J, PL/J, and (SJL/J x PL/J)F1 mice. Influence of I-A haplotypes on encephalitogenic epitope of myelin basic protein.</a> J. Immunol. 141, 1143-1149 (1988).</li><li>Shaw, M. K., Kim, C., Ho, K. -. L., Lisak, R. P., Tse, H. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+combination+of+adoptive+transfer+and+antigenic+challenge+induces+consistent+murine+experimental+autoimmune+encephalomyelitis+in+C57BL/6+mice+and+other+reputed+resistant+strains.">A combination of adoptive transfer and antigenic challenge induces consistent murine experimental autoimmune encephalomyelitis in C57BL/6 mice and other reputed resistant strains.</a> J. Neuroimmunol. 39, 139-150 (1993).</li><li>Krishnamoorthy, G., Saxena, A., Mars, L. T., Domingues, H. S., Mentele, R., Ben-Nun, A., Lassmann, H., Dornmair, K., Kurschus, F. C., Liblau, R. S., Wekerle, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Myelin-specific+T+cells+also+recognize+neuronal+autoantigen+in+a+transgenic+mouse+model+of+multiple+sclerosis.">Myelin-specific T cells also recognize neuronal autoantigen in a transgenic mouse model of multiple sclerosis.</a> Nat. Med. 15, 626-632 (2009).</li><li>Anderson, A. C., Reddy, J., Nazareno, R., Sobel, R. A., Nicholson, L. B., Kuchroo, V. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=IL-10+plays+an+important+role+in+the+homeostatic+regulation+of+the+autoreactive+reperteroire+in+na&#239;ve+mice.">IL-10 plays an important role in the homeostatic regulation of the autoreactive reperteroire in na&#239;ve mice.</a> J. Immunol. 173, 828-834 (2004).</li><li>Anderton, S. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Treg+and+T-effector+cells+in+autoimmune+CNS+inflammation:+a+delicate+balance+easily+disturbed.">Treg and T-effector cells in autoimmune CNS inflammation: a delicate balance easily disturbed.</a> Eur. J. Immunol. 40, 332-334 (2010).</li><li>Arnon, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Experimental+allergic+encephalomyelitis+-+susceptibility+and+suppression.">Experimental allergic encephalomyelitis - susceptibility and suppression.</a> Immunol. Rev. 55, 5 (1981).</li><li>Billiau, A., Heremans, H., Vandekerckhove, F., Dijkmans, R., Sobis, H., Meulepas, E., Carton, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Enhancement+of+experimental+allergic+encephalomyelitis+in+mice+by+antibodies+against+IFN-g.">Enhancement of experimental allergic encephalomyelitis in mice by antibodies against IFN-g.</a> J. Immunol. 140, 1506-1510 (1988).</li><li>Arbomson-Leeman, S., Alexander, J., Bronson, R., Corroll, J., Southwood, S., Dorf, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Experimental+autoimmune+encephalomyelitis-resistant+mice+have+highly+encephalitogenic+myelin+basic+protein+(MBP)-specific+T+cell+clones+that+recognize+a+MBP+peptide+with+high+affinity+for+MHC+class+II.">Experimental autoimmune encephalomyelitis-resistant mice have highly encephalitogenic myelin basic protein (MBP)-specific T cell clones that recognize a MBP peptide with high affinity for MHC class II.</a> J. Immunol. 154, 388-398 (1995).</li><li>Reddy, J., Illes, Z., Zhang, X., Encinas, J., Nicholson, L., Sobel, R. A., Wucherpfennig, K. W., Kuchroo, V. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Myelin+proteolipid+protein-specific+CD4+CD25++regulatory+cells+mediate+genetic+resistance+to+experimental+autoimmune+encephalomyelitis.">Myelin proteolipid protein-specific CD4+CD25+ regulatory cells mediate genetic resistance to experimental autoimmune encephalomyelitis.</a> Proc. Natl. Acad. Sci. U.S.A. 101, 15434-15439 (2004).</li><li>Korn, T., Reddy, J., Gao, W., Bettelli, E., Awasthi, A., Petersen, T. R., Backstrom, B. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Myelin-+specific+regulatory+T+cells+accumulate+in+the+CNS+but+fail+to+control+autoimmune+inflammation.">Myelin- specific regulatory T cells accumulate in the CNS but fail to control autoimmune inflammation.</a> Nature Med. 13, 423-431 (2007).</li><li>Shaw, M. K., Li, J., Ho, P. P., Hao, H., Lisak, R. P., Tse, H. Y., Broglie, P. V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Induction+of+myelin+basic+protein-specific+adoptive+experimental+autoimmune+encephalomyelitis+in+C57BL/6+mice.+Longevity+of+donor+T+cells+and+lack+of+disease+relapses.">Induction of myelin basic protein-specific adoptive experimental autoimmune encephalomyelitis in C57BL/6 mice. Longevity of donor T cells and lack of disease relapses.</a> Neuroimmunology Research Focus. , 175-191 (2007).</li></ol>

لا الإفصاحات المالية.

دراسات من EAE في الفئران غالبا ما تستخدم في neuroantigens، المايلين الأساسية البروتين (MBP)، بروتين شحم بروتيني (حزب العمال التقدمي)، أو بروتين النخاعين oligodendrocyte (موج). دراسات سابقة تستخدم في الغالب MBP. حزب العمال التقدمي يحفز استجابات قوية وثابتة من الفئران SJL. وفي الآونة الأخي?...

<table border="1" style=";text-align:right;direction:rtl"><tbody><tr><td> اسم كاشف </td><td> شركة </td><td> فهرس العدد </td><td> تعليقات </td></tr><tr><td> خنزير غينيا النخاع الشوكي </td><td> روكلاند المناعية للمواد الكيميائية، وشركة Gilbertsville، PA 19525 <a href="http://www.rockland-inc.com" target="_blank">www.rockland-inc.com</a> </td><td> GP-T065 </td><td> تجميد </td></tr><tr><td> مساند فرويند، H37Ra كاملة </td><td> Difco مختبرات ديترويت، ميتشيغن </td><td> 231131 </td><td></td></tr><tr><td> Multifit تبادل، وقادر على محقنة </td><td> دينار بحريني <a href="http://www.bd.com/us" target="_blank">www.bd.com / لنا</a> </td><td> 512133 </td><td></td></tr><tr><td> RPMI 1640 </td><td> Mediatechالمؤتمر الوطني العراقي. ماناساس، فرجينيا <a href="http://www.cellgro.com" target="_blank">www.cellgro.com</a> </td><td> 10-040-CM </td><td></td></tr><tr><td> OVA </td><td> سيغما الدريخ <a href="http://www.sigmaaldrich.com" target="_blank">www.sigmaaldrich.com</a> </td><td> A5503 </td><td></td></tr><tr><td> الفئران </td><td> مختبر جاكسون بار هاربور، مين </td><td> B6 الفئران: 0000664 </td><td></td></tr></tbody></table>

overcoming unresponsiveness in experimental autoimmune encephalomyelitis (eae) resistant mouse strains by adoptive transfer and antigenic challenge

التهاب الدماغ والنخاع الذاتية التجريبية (EAE) هو مرض التهاب الجهاز العصبي المركزي (CNS)، وقد استخدمت في نموذج حيواني لدراسة هذا المرض المزيل للبشرية، والتصلب المتعدد (MS). ويتميز EAE بواسطة تسلل مرضية للخلايا وحيدات النوى في الجهاز العصبي المركزي والمظاهر السريرية للمرض مشلول. مشابهة لمرض التصلب العصبي المتعدد، EAE هو أيضا تحت السيطرة الوراثية في ذلك سلالات الفئران معينة تكون عرضة للتحريض المرض والبعض الآخر مقاومة. عادة، C57BL / 6 (H-2 ب) تحصين الفئران مع البروتين الأساسي المايلين (MBP) فشل لتطوير علامات مشلول. هذا تجاوب هي بالتأكيد ليست بسبب عيوب في التصنيع أو مستضد تقديم المستضد من MBP، كما تم استخدام بروتوكول تجريبي وصفها هنا للحث على EAE حاد في C57BL / 6 الفئران فضلا عن غيرها من سمعته الطيبة سلالات الفئران المقاومة. وبالإضافة إلى ذلك، قد دماغي المنشأ استنساخ الخلايا التائية من C57BL / 6 و BALB / ج الفئران على رد الفعل إلى MBP كان نجاحمعزولة تماما وترويجها. بروتوكول تجريبي ينطوي على استخدام الخلوي نظام نقل بالتبني الذي MBP-معبي (200 ميكروغرام / الماوس) معزولة C57BL / 6 خلايا العقدة الليمفاوية المانحة وتربيتها لمدة خمسة أيام مع مستضد إلى توسيع مجموعة خلايا تي MBP محددة. في نهاية الفترة والثقافة، ويتم نقل 50000000 خلايا قابلة للحياة إلى المتلقين مسانج ساذج عن طريق الوريد الذيل. الفئران المعالجة لذلك المتلقي عادة لا تتطور EAE، مما يؤكد وضعهم المقاومة، وأنها يمكن أن تبقى إلى أجل غير مسمى طبيعي. عشرة أيام نقل الخلية آخر، هي الطعن الفئران المتلقي مع مساعد فرويند الكامل (CFA)، مستحلب MBP في أربعة مواقع في جنباته. EAE حاد يبدأ في تطوير هذه الفئران 10-14 يوما بعد التحدي. وأظهرت النتائج أن الاستقراء من مرض معين كان المستضدية كما تحد مع مستضدات غير ذي صلة لم لحث على العلامات السريرية للمرض. بشكل ملحوظ، والمعايرة من الجرعة المستخدمة لمستضدتحدي أظهرت الفئران المتلقي أنه يمكن أن يكون ما يصل الى 5 ميكروغرام / الماوس. وبالإضافة إلى ذلك، أظهرت دراسة الحركية للتوقيت التحدي مستضدي هذا التحدي للحث على المرض كانت فعالة في أقرب وقت آخر 5 أيام التحدي المستضدية وطالما أن أكثر من 445 يوما آخر مستضدي التحدي. هذه البيانات نقطة بقوة نحو تورط &quot;طويلة الأمد&quot; سكان الخلية تي في الحفاظ على عدم استجابتها. لم يتم تعريف اشتراك الخلايا التائية التنظيمية (Tregs) في هذا النظام.

Watch this Scientific Journal Video about Overcoming Unresponsiveness in Experimental Autoimmune Encephalomyelitis EAE Resistant Mouse Strains by Adoptive Transfer and Antigenic Challenge at JoVE.com

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

سلالات الفئران معينة قادرة على مقاومة تحريض التهاب الدماغ والنخاع الذاتية التجريبية (EAE) مع البروتين الأساسي المايلين. الموصوفة هنا هو بروتوكول تحصين بسيط هو أن يعكس عدم تجاوب ويستحث مرض شلل في نموذجي EAE عدة بقع الفأر المقاوم.

التغلب على عدم التجاوب في التجارب الذاتية التهاب الدماغ (EAE) سلالات الفئران المقاومة للنقل بواسطة بالتبني والتحدي الأنتيجين

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

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Immunology and Infection

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

20.2K Views.  St. John-Providence Health System. سلالات الفئران معينة قادرة على مقاومة تحريض التهاب الدماغ والنخاع الذاتية التجريبية (EAE) مع البروتين الأساسي المايلين. الموصوفة هنا هو بروتوكول تحصين بسيط هو أن يعكس عدم تجاوب ويستحث مرض شلل في نموذجي EAE عدة بقع الفأر المقاوم.

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

Experimental Metastasis and CTL Adoptive Transfer Immunotherapy Mouse Model

Experimental metastasis mouse model is a simple and yet physiologically relevant metastasis model. The tumor cells are injected intravenously (i.v) into mouse tail veins and colonize in the lungs, thereby, resembling the last steps of tumor cell spontaneous metastasis: survival in the circulation, extravasation and colonization in the distal organs. From a therapeutic point of view, the experimental metastasis model is the simplest and ideal model since the target of therapies is often the end point of metastasis: established metastatic tumor in the distal organ. In this model, tumor cells are injected i.v into mouse tail veins and allowed to colonize and grow in the lungs. Tumor-specific CTLs are then injected i.v into the metastases-bearing mouse. The number and size of the lung metastases can be controlled by the number of tumor cells to be injected and the time of tumor growth. Therefore, various stages of metastasis, from minimal metastasis to extensive metastasis, can be modeled. Lung metastases are analyzed by inflation with ink, thus allowing easier visual observation and quantification.

An experimental lung metastasis and CTL immunotherapy mouse model for analysis of tumor cells-T cell interaction in vivo.

الإنبثاث التجريبية وCTL المناعي نقل بالتبني النموذجي ماوس

Experimental metastasis mouse model is a simple and yet physiologically relevant metastasis model. The tumor cells are injected intravenously (i.v) into ...

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.

تحريض التهاب النخامية الذاتية التجريبية في الفئران SJL

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

Accelerated Type 1 Diabetes Induction in Mice by Adoptive Transfer of Diabetogenic CD4+ T Cells

The nonobese diabetic (NOD) mouse spontaneously develops autoimmune diabetes after 12 weeks of age and is the most extensively studied animal model of human Type 1 diabetes (T1D). Cell transfer studies in irradiated recipient mice have established that T cells are pivotal in T1D pathogenesis in this model. We describe herein a simple method to rapidly induce T1D by adoptive transfer of purified, primary CD4+ T cells from pre-diabetic NOD mice transgenic for the islet-specific T cell receptor (TCR) BDC2.5 into NOD.SCID recipient mice. The major advantages of this technique are that isolation and adoptive transfer of diabetogenic T cells can be completed within the same day, irradiation of the recipients is not required, and a high incidence of T1D is elicited within 2 weeks after T cell transfer. Thus, studies of pathogenesis and therapeutic interventions in T1D can proceed at a faster rate than with methods that rely on heterogenous T cell populations or clones derived from diabetic NOD mice.

We provide a reproducible method to induce type 1 diabetes (T1D) in mice within two weeks by the adoptive transfer of islet antigen-specific, primary CD4+ T cells.

المعجل نوع 1 الحث السكري في الفئران عن طريق نقل بالتبني من CD4 + T خلايا سكري

The nonobese diabetic (NOD) mouse spontaneously develops autoimmune diabetes after 12 weeks of age and is the most extensively studied animal model of ...

Myelin Oligodendrocyte Glycoprotein (MOG35-55) Induced Experimental Autoimmune Encephalomyelitis (EAE) in C57BL/6 Mice

Multiple sclerosis is a chronic neuroinflammatory demyelinating disorder of the central nervous system with a strong neurodegenerative component. While the exact etiology of the disease is yet unclear, autoreactive T lymphocytes are thought to play a central role in its pathophysiology. MS therapy is only partially effective so far and research efforts continue to expand our knowledge on the pathophysiology of the disease and to develop novel treatment strategies. Experimental autoimmune encephalomyelitis (EAE) is the most common animal model for MS sharing many clinical and pathophysiological features. There is a broad diversity of EAE models which reflect different clinical, immunological and histological aspects of human MS. Actively-induced EAE in mice is the easiest inducible model with robust and replicable results. It is especially suited for investigating the effects of drugs or of particular genes by using transgenic mice challenged by autoimmune neuroinflammation. Therefore, mice are immunized with CNS homogenates or peptides of myelin proteins. Due to the low immunogenic potential of these peptides, strong adjuvants are used. EAE susceptibility and phenotype depends on the chosen antigen and rodent strain. C57BL/6 mice are the commonly used strain for transgenic mouse construction and respond among others to myelin oligodendrocyte glycoprotein (MOG). The immunogenic epitope MOG35-55 is suspended in complete Freund's adjuvant (CFA) prior to immunization and pertussis toxin is applied on the day of immunization and two days later. Mice develop a "classic" self-limited monophasic EAE with ascending flaccid paralysis within 9-14 days after immunization. Mice are evaluated daily using a clinical scoring system for 25-50 days. Special considerations for care taking of animals with EAE as well as potential applications and limitations of this model are discussed.

Myelin Oligodendrocyte Glycoprotein MOG35-55 Induced Experimental Autoimmune Encephalomyelitis EAE in C57BL/6 Mice

Experimental autoimmune encephalomyelitis (EAE) is an established animal model of multiple sclerosis. C57BL/6 mice are immunized with myelin oligodendrocyte glycoprotein (MOG) peptide 35-55 (MOG35-55), resulting in an ascending flaccid paralysis caused by autoreactive immune cells in the central nervous system. Protocols for disease induction and monitoring will be discussed.

المايلين دبقية قليلة التغصن جلايكوبروتين (MOG<sub&gt; 35-55</sub&gt;) المستحثة التجريبية المناعة الذاتية التهاب الدماغ (EAE) في C57BL / 6 الفئران

Multiple sclerosis is a chronic neuroinflammatory demyelinating disorder of the central nervous system with a strong neurodegenerative component. While ...

Induction of Murine Intestinal Inflammation by Adoptive Transfer of Effector CD4+CD45RBhigh T Cells into Immunodeficient Mice

There are many different animal models available for studying the pathogenesis of human inflammatory bowel diseases (IBD), each with its own advantages and disadvantages. We describe here an experimental colitis model that is initiated by adoptive transfer of syngeneic splenic CD4+CD45RBhigh T cells into T and B cell deficient recipient mice. The CD4+CD45RBhigh T cell population that largely consists of na&#239;ve effector cells is capable of inducing chronic intestinal inflammation, closely resembling key aspects of human IBD. This method can be manipulated to study aspects of disease onset and progression. Additionally it can be used to study the function of innate, adaptive, and regulatory immune cell populations, and the role of environmental exposures, i.e., the microbiota, in intestinal inflammation. In this article we illustrate the methodology for inducing colitis with a step-by-step protocol. This includes a video demonstration of key technical aspects required to successfully develop this murine model of experimental colitis for research purposes.

Induction of Murine Intestinal Inflammation by Adoptive Transfer of Effector CD4+CD45RBhigh T Cells into Immunodeficient Mice

Here, we present a protocol to induce colonic inflammation in mice by adoptive transfer of syngeneic CD4+CD45RBhigh T cells into T and B cell deficient recipients. Clinical and histopathological features mimic human inflammatory bowel diseases. This method allows the study of the initiation of colonic inflammation and progression of disease.

تحريض الفئران المعوية التهاب عن طريق نقل بالتبني من المستجيب CD4<sup&gt; +</sup&gt; CD45RB<sup&gt; عالية</sup&gt; خلايا T في الفئران العوز المناعي

There are many different animal models available for studying the pathogenesis of human inflammatory bowel diseases (IBD), each with its own advantages ...

Induction of Experimental Autoimmune Encephalomyelitis in Mice and Evaluation of the Disease-dependent Distribution of Immune Cells in Various Tissues

Multiple sclerosis is presumed to be an inflammatory autoimmune disease, which is characterized by lesion formation in the central nervous system (CNS) resulting in cognitive and motor impairment. Experimental autoimmune encephalomyelitis (EAE) is a useful animal model of MS, because it is also characterized by lesion formation in the CNS, motor impairment and is also driven by autoimmune and inflammatory reactions. One of the EAE models is induced with a peptide derived from the myelin oligodendrocyte protein (MOG)35-55 in mice. The EAE mice develop a progressive disease course. This course is divided into three phases: the preclinical phase (day 0 - 9), the disease onset (day 10 - 11) and the acute phase (day 12 - 14). MS and EAE are induced by autoreactive T cells that infiltrate the CNS. These T cells secrete chemokines and cytokines which lead to the recruitment of further immune cells. Therefore, the immune cell distribution in the spinal cord during the three disease phases was investigated. To highlight the time point of the disease at which the activation/proliferation/accumulation of T cells, B cells and monocytes starts, the immune cell distribution in lymph nodes, spleen and blood was also assessed. Furthermore, the levels of several cytokines (IL-1&#946;, IL-6, IL-23, TNF&#945;, IFN&#947;) in the three disease phases were determined, to gain insight into the inflammatory processes of the disease. In conclusion, the data provide an overview of the functional profile of immune cells during EAE pathology.

This manuscript describes the methods for induction and scoring of the experimental autoimmune encephalomyelitis (EAE) model, together with the assessment of immune cell distribution and mRNA cytokine levels in lymph nodes, spleen, blood and spinal cord using flow cytometry and quantitative PCR, respectively, at various disease phases.

تحريض التجريبية المناعة الذاتية التهاب الدماغ في الفئران وتقييم توزيع الأمراض التي تعتمد على خلايا المناعة في الأنسجة المختلفة

Multiple sclerosis is presumed to be an inflammatory autoimmune disease, which is characterized by lesion formation in the central nervous system (CNS) ...

Simple and Efficient Production and Purification of Mouse Myelin Oligodendrocyte Glycoprotein for Experimental Autoimmune Encephalomyelitis Studies

Multiple sclerosis (MS) is a chronic inflammatory disease of the central nervous system (CNS), thought to occur as a result of autoimmune responses targeting myelin. Experimental autoimmune encephalomyelitis (EAE) is the most common animal model of CNS autoimmune disease, and is typically induced via immunization with short peptides representing immunodominant CD4+ T cell epitopes of myelin proteins. However, B cells recognize unprocessed protein directly, and immunization with short peptide does not activate B cells that recognize the native protein. As recent clinical trials of B cell-depleting therapies in MS have suggested a role for B cells in driving disease in humans, there is an urgent need for animal models that incorporate B cell-recognition of autoantigen. To this end, we have generated a new fusion protein containing the extracellular domain of the mouse version of myelin oligodendrocyte glycoprotein (MOG) as well as N-terminal fusions of a His-tag for purification purposes and the thioredoxin protein to improve solubility (MOGtag). A tobacco etch virus (TEV) protease cleavage site was incorporated to allow the removal of all tag sequences, leaving only the pure MOG1-125 extracellular domain. Here, we describe a simple protocol using only standard laboratory equipment to produce large quantities of pure MOGtag or MOG1-125. This protocol consistently generates over 200 mg of MOGtag protein. Immunization with either MOGtag or MOG1-125 generates an autoimmune response that includes pathogenic B cells that recognize the native mouse MOG.

We describe a simple protocol using only basic lab equipment to generate and purify large quantities of a fusion protein that contains mouse Myelin Oligodendrocyte Glycoprotein. This protein can be used to induce experimental autoimmune encephalomyelitis driven by both T and B cells.

إنتاج بسيطة وفعالة وتنقية ماوس المايلين دبقية قليلة التغصن البروتين السكري لالتجريبية المناعة الذاتية الدراسات التهاب الدماغ

Multiple sclerosis (MS) is a chronic inflammatory disease of the central nervous system (CNS), thought to occur as a result of autoimmune responses ...

Visualizing Impairment of the Endothelial and Glial Barriers of the Neurovascular Unit during Experimental Autoimmune Encephalomyelitis In Vivo

The neurovascular unit (NVU) is composed of microvascular endothelial cells forming the blood-brain barrier (BBB), an endothelial basement membrane with embedded pericytes, and the glia limitans composed by the parenchymal basement membrane and astrocytic end-feed embracing the abluminal aspect of central nervous system (CNS) microvessels. In addition to maintaining CNS homeostasis the NVU controls immune cell trafficking into the CNS. During immunosurveillance of the CNS low numbers of activated lymphocytes can cross the endothelial barrier without causing BBB dysfunction or clinical disease. In contrast, during neuroinflammation such as in multiple sclerosis or its animal model experimental autoimmune encephalomyelitis (EAE) a large number of immune cells can cross the BBB and subsequently the glia limitans eventually reaching the CNS parenchyma leading to clinical disease. 
Immune cell migration into the CNS parenchyma is thus a two-step process that involves a sequential migration across the endothelial and glial barrier of the NVU employing distinct molecular mechanisms. If following their passage across the endothelial barrier, T cells encounter their cognate antigen on perivascular antigen-presenting cells their local reactivation will initiate subsequent mechanisms leading to the focal activation of gelatinases, which will enable the T cells to cross the glial barrier and enter the CNS parenchyma. Thus, assessing both, BBB permeability and MMP activity in spatial correlation to immune cell accumulation in the CNS during EAE allows to specify loss of integrity of the endothelial and glial barriers of the NVU. 
We here show how to induce EAE in C57BL/6 mice by active immunization and how to subsequently analyze BBB permeability in vivo using a combination of exogenous fluorescent tracers. We further show, how to visualize and localize gelatinase activity in EAE brains by in situ zymogaphy coupled to immunofluorescent stainings of BBB basement membranes and CD45+ invading immune cells.

Here, we present protocols to investigate impairment of the neurovascular unit during experimental autoimmune encephalomyelitis in vivo. We specifically address how to determine blood-brain barrier permeability and gelatinase activity involved in leukocyte migration across the glia limitans.&#160;

تصور ضعف الحواجز بطانية والدبقيه نيوروفاسكولار وحدة خلال النخاع الذاتية التجريبية في فيفو

The neurovascular unit (NVU) is composed of microvascular endothelial cells forming the blood-brain barrier (BBB), an endothelial basement membrane with ...

Adoptive Immunotherapy of iNKT Cells in Glucose-6-Phosphate Isomerase (G6PI)-Induced RA Mice

Rheumatoid arthritis (RA) is a complex chronic inflammatory autoimmune disease. The pathogenesis of the disease is related to invariant natural killer T (iNKT) cells. Patients with active RA present fewer iNKT cells, defective cell function, and excessive polarization of Th1. In this study, an RA animal model was established using a mixture of hGPI325-339 and hGPI469-483 peptides. The iNKT cells were obtained by in vivo induction and in vitro purification, followed by infusion into RA mice for adoptive immunotherapy. The in vivo imaging system (IVIS) tracking revealed that iNKT cells were mainly distributed in the spleen and liver. On day 12 after cell therapy, the disease progression slowed down significantly, the clinical symptoms were alleviated, the abundance of iNKT cells in the thymus increased, the proportion of iNKT1 in the thymus decreased, and the levels of TNF-&#945;, IFN-&#947;, and IL-6 in the serum decreased. Adoptive immunotherapy of iNKT cells restored the balance of immune cells and corrected the excessive inflammation of the body.

Adoptive Immunotherapy of iNKT Cells in Glucose-6-Phosphate Isomerase G6PI-Induced RA Mice

This protocol uses G6PI mixed peptides to construct rheumatoid arthritis models that are closer to that of human rheumatoid arthritis in CD4+ T cells and cytokines. High purity invariant natural killer T cells (mainly iNKT2) with specific phenotypes and functions were obtained by in vivo induction and in vitro purification for adoptive immunotherapy.

العلاج المناعي بالتبني من خلايا iNKT في الجلوكوز-6-الفوسفات Isomerase (G6PI) الناجمة عن الفئران RA

Rheumatoid arthritis (RA) is a complex chronic inflammatory autoimmune disease. The pathogenesis of the disease is related to invariant natural killer T ...

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.

تحليل تدفق Cytometric لتسلل الخلايا اللمفاوية في الجهاز العصبي المركزي أثناء التهاب الدماغ المناعي الذاتي التجريبي

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

التغلب على عدم التجاوب في التجارب الذاتية التهاب الدماغ (EAE) سلالات الفئران المقاومة للنقل بواسطة بالتبني والتحدي الأنتيجين

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