نحن مدينون لنانسي ر. شتورم، قسم علم المناعة، علم الأحياء الدقيقة والبيولوجيا الجزيئية، وديفيد جيفن في كلية الطب، جامعة كاليفورنيا في لوس انجليس، لقراءة نقدية للمخطوطة. وأيد المجلس الوطني للبحوث، CNPq، ومؤسسة البحوث للتنمية، FAPDF، والبرازيل، والدراسة. نشكر المساعدات الفنية من اليساندرو O. سوزا، ماريا Guimaro جيم، كورديرو سيرو، آنا دي روزا كاسيا، ألفيس Roseneide، واندرادي رافائيل، من جامعة برازيليا، البرازيل.

<ol>
	<li>Teixeira, A. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pathogenesis+of+chagas'+disease:+parasite+persistence+and+autoimmunity.">Pathogenesis of chagas' disease: parasite persistence and autoimmunity.</a> CMR. 24, 592-630 (2011).</li><li>Teixeira, A. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chagas+disease.">Chagas disease.</a> Postg. Med. J. 82, 788-798 (2006).</li><li>Teixeira, A. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Trypanosoma+cruzi+in+the+chicken+model:+Chagas-like+heart+disease+in+the+absence+of+parasitism.">Trypanosoma cruzi in the chicken model: Chagas-like heart disease in the absence of parasitism.</a> PLoS Negl. Trop. Dis. 5, e1000 (2011).</li><li>Hecht, M. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Inheritance+of+DNA+transferred+from+American+trypanosomes+to+human+hosts.">Inheritance of DNA transferred from American trypanosomes to human hosts.</a> PLoS One. 5, e9181 (2010).</li><li>Xing, Z. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Roles+of+the+ERK+MAPK+in+the+regulation+of+proinflammatory+and+apoptotic+responses+in+chicken+macrophages+infected+with+H9N2+avian+influenza+virus.">Roles of the ERK MAPK in the regulation of proinflammatory and apoptotic responses in chicken macrophages infected with H9N2 avian influenza virus.</a> J. Gen. Virol. 91, 343-351 (2010).</li><li>Kim, H. 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R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sensitive+detection+and+schizodeme+classification+of+Trypanosoma+cruzi+cells+by+amplification+of+kinetoplast+minicircle+DNA+sequences:+use+in+diagnosis+of+Chagas'+disease.">Sensitive detection and schizodeme classification of Trypanosoma cruzi cells by amplification of kinetoplast minicircle DNA sequences: use in diagnosis of Chagas' disease.</a> Mol. Biochem. Parasitol. 33, 205-214 (1989).</li><li>Teixeira, A. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evolution+and+pathology+in+chagas+disease--a+review.">Evolution and pathology in chagas disease--a review.</a> Mem. Inst. Oswaldo Cruz. 101, 463-491 (2006).</li><li>Yurchenko, V. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Structure+of+Leishmania+minicircle+kinetoplast+DNA+classes.">Structure of Leishmania minicircle kinetoplast DNA classes.</a> J. Clin. Microbiol. 37, 1656-1657 (1999).</li><li>Simpson, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+genomic+organization+of+guide+RNA+genes+in+kinetoplastid+protozoa:+several+conundrums+and+their+solutions.">The genomic organization of guide RNA genes in kinetoplastid protozoa: several conundrums and their solutions.</a> Mol. Biochem. Parasitol. 86, 133-141 (1997).</li><li>Thomas, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+non-universal+transcription+factor?+The+Leishmania+tarentolae+TATA+box-binding+protein+LtTBP+associates+with+a+subset+of+promoters.">A non-universal transcription factor? The Leishmania tarentolae TATA box-binding protein LtTBP associates with a subset of promoters.</a> Int J. Parasitol. 36, 1217-1226 (2006).</li></ol>

الإعلان عن أي تضارب في المصالح.

وعلى النقيض من الثدييات عرضة لمدى الحياة ت. التهابات الكروزية، دجاجة وهي لصهر إلى T. الكروزية عدوى. والميزة الرئيسية لنظام الدجاج النموذج هو القضاء على هذا المرض في وقت مبكر في تطوير الجهاز المناعي الجنينية. وبالتالي، تم دمج الحمض النووي طفيلي الوحيدة المتبقية داخل الجسم الدجاج في مواضع عدة. استخدم كمية الأمثل للT. ضراوة الكروزية trypomastigotes لتطعيم البيضة الخصبة هو خطوة حاسمة نحو الحصول على التكامل بين minicircles kDNA في الدجاج جينوم الجنين. معدل فقس بيض الدجاج الحي من تلقيح مع 100 trypomastigotes هو أربعة أضعاف أعلى من تلك التي تم الحصول عليها مع 500 الطفيليات. ينبغي الحرص على تطعيم تعليق طفيليات في 10 ميكرولتر من مستنبت داخل البويضة غرفة الهواء. وينبغي أن يكون هناك أي تسرب من بياض البيض. في الظروف المثالية، وإصابة الخلايا الطفيلية يأخذ واي مكانرقيق بعد بضع ساعات من الضرب وحضانة طفيلي داخل الخلايا المضيفة العائدات لمدة أسبوع، بعد ذلك يتم التخلص من العدوى من المناعة الفطرية. التكامل kDNA يتطلب عدوى المعيشة، وتلقيح minicircles عارية في وقت مبكر بيض الدجاج الجنين لا يؤدي إلى التكامل. وينبغي أن يضم الأجنة kDNA إيجابية والضوابط تحت ظروف خاضعة للرقابة على 37،5 درجة مئوية والرطوبة 65٪. يتم الاحتفاظ في أقفاص الدجاج لمدة أسبوعين عند 33 درجة مئوية درجة حرارة الغرفة. بعد ذلك يتم الاحتفاظ بها في أقفاص الدجاج على رفوف معلقة فصل بمقدار 1.5 متر في عرض الممرات غرفة في 22 درجة مئوية مع الهواء التي تمت تصفيتها وتحت ضغط إيجابي استنفاد مستمر لتأمين ظروف رعاية الحيوان. ويتم تغذية الكبار الدجاج طعام والشراب شرب المياه الجارية لتحقيق النمو والنضج الكامل، وضع البيض في خمسة أشهر من العمر. صيانة الإجراءات الصحية ضرورية لاستنساخ نتائج عند العمل مع ت. الكروزية تلقيحفي بيض الدجاج الخصبة. في الدجاج نظام النموذج T. يتم القضاء على العدوى الكروزية بعد تطوير نظام المناعة في الفترة المبكرة من نمو الجنين. وبالإضافة إلى ذلك إلى كونها خالية من العدوى، والدجاج الذي يفقس من T. الكروزية تلقيح البيض، في عدم وجود أجسام مضادة محددة، ومتسامحون للمستضدات الطفيلي. ويعتبر رفض خلايا القلب الهدف بواسطة الخلايا اللمفية السامة للخلايا (الحد الأدنى من وحدة الرفض، الشكل 3) في دجاجة kDNA-تحور تظهر التعديلات التركيب الوراثي وانهيار المراقبة المناعية 3. وتعديل genotypically الخلايا التائية رفض تقديم المتسارع للأنسجة النفس في الجسم. الموقع آفة الرئيسي هو القلب، والذي هو السمة المميزة لمرض شاغاس. ويعتبر الممر من الفسيولوجية (المراقبة) لدولة فيزيوباثي في الدجاج kDNA-تحور تظهر clonally انتشار الخلايا اللمفية السامة للخلايا 3. تينظام له نموذج transkingdom يظهر الطفيلي التي يسببها، وراثيا يحركها أمراض المناعة الذاتية (الشكل 6)، الناجمة عن التعديلات الوراثية التي ت. الكروزية kDNA minicircle التكامل. لا ينظر إلى هذه التعديلات في الكتاكيت فقس البيض من رطل تلقيح. هذه الظاهرة تشير إلى أن علاجا تجريبيا من اعتلال عضلة القلب والتهابات المناعة الذاتية في kDNA-تحور الدجاج قد تتطلب قمع المخدرات من السلف نخاع العظم من النمط الظاهري تي خلية محددة التسلل الى عضلة القلب، وزرع نخاع العظم متوافق نسيجيا صحي لمنع رفض الأنسجة الذاتي.

<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> طق بوليميريز الحمض النووي المؤتلف </td><td> إينفيتروجن </td><td> 11615-010 </td><td></td></tr><tr><td> البوليميراز البلاتين طق الحمض النووي </td><td> إينفيتروجن </td><td> 10966-030 </td><td></td></tr><tr><td> عشوائية وصفها DNA نظام الاشعال </td><td> إينفيتروجن </td><td> 18187-013 </td><td></td></tr><tr><td> الايكولوجية RI </td><td> إينفيتروجن </td><td> 15202-021 </td><td></td></tr><tr><td> أنا مبو </td><td> إينفيتروجن </td><td> 15248-016 </td><td></td></tr><tr><td> dNTP مجموعة، حلول 100mM </td><tد&gt; شركة جنرال الكتريك للرعاية الصحية </td><td> 28-4065-51 </td><td></td></tr><tr><td> أمرشام Hybond - N + - ن قطة. </td><td> جنرال إلكتريك للرعاية الصحية </td><td> RPN303B </td><td></td></tr><tr><td> PlasmidPrep البسيطة سبين عدة </td><td> جنرال إلكتريك للرعاية الصحية </td><td> 28-9042-70 </td><td></td></tr><tr><td> NSI أنا </td><td> SIGMA الدريخ </td><td> R5884 1KU </td><td></td></tr><tr><td> الحمض النووي، والسمك ملح الصوديوم الحيوانات المنوية </td><td> AMRESCO </td><td> 0644-10G </td><td></td></tr><tr><td> الماوس المضادة للدجاج بو-1B </td><td> SouthernBiotech </td><td> 8370-02 </td><td></td></tr><tr><td> الماوس المضادة للدجاج CD45 </td><td> SouthernBiotech </td><td> 8270-08 </td><td></td></tr><tr><td>الماوس المضادة للدجاج TCRγδ </td><td> SouthernBiotech </td><td> 8230-08 </td><td></td></tr><tr><td> الماوس المضادة للدجاج CD8α </td><td> SouthernBiotech </td><td> 9220-02 </td><td></td></tr><tr><td> الماوس المضادة للدجاج الوحيدات / بلعم </td><td> SouthernBiotech </td><td> 8420-02 </td><td></td></tr><tr><td> MyCycle Termocycler </td><td> الحيوي راد مختبرات </td><td> 580BR 5501 </td><td></td></tr></tbody></table>

parasite induced genetically driven autoimmune chagas heart disease in the chicken model

والكروزية الالتهابات الحادة المثقبية المكتسبة في سن الرضاعة والطفولة تبدو أعراض، ولكن ما يقرب من ثلث الحالات المصابة بشكل مزمن تظهر داء شاغاس تصل إلى ثلاثة عقود أو في وقت لاحق. المناعة الذاتية والمثابرة طفيلي تتنافس النظريات لتفسير الآلية المرضية لداء شاغاس 1 و 2. إلى أدوار منفصلة لعبت من قبل استمرار طفيلي والمناعة الذاتية في مرض شاغاس نحن تطعيم في T. الكروزية في غرفة الهواء من البويضات المخصبة. النظام المناعي الدجاج ناضج هو حاجز ضيق البيولوجية ضد ت. يتم القضاء على الكروزية والعدوى وعلى تطوير نظام المناعة في نهاية الأسبوع الأول من نمو 3. الكتاكيت هي خالية من طفيلي عند الفقس، ولكنها تحتفظ دمجها طفيلي الميتوكوندريا kinetoplast الحمض النووي (kDNA) minicircle ضمن الجينوم الخاصة بهم التي يتم نقلها إلى ذريتها. تم الحصول على وثائق من التكامل kDNA minicircle في جينوم الدجاج من قبل targeted رئيس الذيل PCR، التهجين الجنوبية، والاستنساخ، والتسلسل 3 و 4. وkDNA minicircle التكامل تمزق الإطارات القراءة مفتوحة للنسخ والعوامل الجهاز المناعي، والفوسفاتيز (GTPase)، محلقة الأدينيلات وphosphorylases (بي كي سي، NF-B كابا المنشط، PI-3K) المرتبطة فسيولوجيا الخلية، والنمو، والتمايز 3، 5 - 7 وظائف الجينات الأخرى. ويعتبر التهاب عضلة القلب شديدة بسبب الرفض من ألياف قلب الهدف بواسطة الخلايا اللمفية المستجيبات السامة للخلايا في الدجاج kDNA تحور، مبديا التهابات عضلة القلب شبيهة بتلك التي شوهدت في مرض شاغاس الإنسان. وجدير بالذكر أن فشل القلب والهيكل العظمي وضعف العضلات موجودة في الدجاج الكبار مع تمزق kDNA من الجين dystrophin في كروموسوم 1 8. وترتبط التعديلات genotipic مماثلة مع تدمير الأنسجة التي تقوم بها المستجيبات CD45 +، + CD8γδ، CD8α الخلايا اللمفية. وبالتالي يمكن لهذه العدوى الأوالي حمل أمراض المناعة الذاتية يحركها وراثيا.

Watch this Scientific Journal Video about Parasite Induced Genetically Driven Autoimmune Chagas Heart Disease in the Chicken Model at JoVE.com

Parasite Induced Genetically Driven Autoimmune Chagas Heart Disease in the Chicken Model

وتلقيح<em&gt; المثقبية الكروزية</em&gt; في البيض الخصبة قبل الحضانة يجعل kDNA طفيلي minicircle التكامل في جينوم خلايا الجنين. تهجين يكشف عن نقل عمودي من الطفرات إلى ذرية. وkDNA يدمج الترميز مناطق عدة في الصبغيات (الكروموسومات) والدجاجة تموت مع مرض القلب الذاتية للالتهابات.

المستحث الطفيلي قيادة وراثيا القلب المناعة الذاتية ومرض شاغاس في نموذج الدجاج

The Trypanosoma cruzi acute infections acquired in infancy and childhood seem asymptomatic, but approximately one third of the chronically infected cases ...

parasite-induced-genetically-driven-autoimmune-chagas-heart-disease

Research

JoVE Journal

Immunology and Infection

13.7K Views.  University of Brasilia. وتلقيح المثقبية الكروزية في البيض الخصبة قبل الحضانة يجعل kDNA طفيلي minicircle التكامل في جينوم خلايا الجنين. تهجين يكشف عن نقل عمودي من الطفرات إلى ذرية. وkDNA يدمج الترميز مناطق عدة في الصبغيات (الكروموسومات) والدجاجة تموت مع مرض القلب الذاتية للالتهابات.

Video: Parasite Induced Genetically Driven Autoimmune Chagas Heart Disease in the Chicken Model

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 ...

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 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) ...

Sexual Transmission of American Trypanosomes from Males and Females to Naive Mates

American trypanosomiasis is transmitted to humans by triatomine bugs through the ingestion of contaminated food, by blood transfusions or accidently in hospitals and research laboratories. In addition, the Trypanosoma cruzi infection is transmitted congenitally from a&#160;chagasic mother to her offspring, but the male partner&#39;s contribution to in utero contamination is unknown. The findings of nests and clumps of amastigotes and of trypomastigotes in the theca cells of the ovary, in the goniablasts and in the lumen of seminiferous tubules suggest that T. cruzi infections are sexually transmitted. The research protocol herein presents the results of a family study population showing parasite nuclear DNA in the diploid blood mononuclear cells and in the haploid gametes of human subjects. Thus, three independent biological samples collected one year apart confirmed that T. cruzi infections were sexually transmitted to progeny. Interestingly, the specific T. cruzi antibody was absent in the majority of family progeny that bore immune tolerance to the parasite antigen. Immune tolerance was demonstrated in chicken refractory to T. cruzi after the first week of embryonic growth, and chicks hatched from the flagellate-inoculated eggs were unable to produce the specific antibody.&#160;Moreover, the instillation of the human semen ejaculates intraperitoneally or into the vagina of naive mice yielded T. cruzi amastigotes in the epididymis, seminiferous tubule, vas deferens and uterine tube with an absence of inflammatory reactions in the immune privileged organs of reproduction. The breeding of T. cruzi-infected male and female mice with naive mates resulted in acquisition of the infections, which were later transmitted to the progeny. Therefore, a robust education, information and communication program that involves the population and social organizations is deemed necessary to prevent Chagas disease.

The Trypanosoma cruzi agent of Chagas disease produces long-lasting asymptomatic infections that abruptly develop into clinically recognized pathology. The following research protocol describes a short-run family-based epidemiological study to unravel the T. cruzi infection transmitted sexually from parent to progeny.

الاتصال الجنسي من المثقبيات الأمريكية من الذكور والإناث إلى الأصحاب ساذج

American trypanosomiasis is transmitted to humans by triatomine bugs through the ingestion of contaminated food, by blood transfusions or accidently in ...

An Ex Vivo Chicken Primary Bursal-cell Culture Model to Study Infectious Bursal Disease Virus Pathogenesis

Infectious bursal disease virus (IBDV) is a birnavirus of economic importance to the poultry industry. The virus infects B cells, causing morbidity, mortality, and immunosuppression in infected birds. In this study, we describe the isolation of chicken primary bursal cells from the bursa of Fabricius, the culture and infection of the cells with IBDV, and the quantification of viral replication. The addition of chicken CD40 ligand significantly increased cell proliferation fourfold over six days of culture and significantly enhanced cell viability. Two strains of IBDV, a cell-culture adapted strain, D78, and a very virulent strain, UK661, replicated well in the ex vivo cell cultures. This model will be of use in determining how cells respond to IBDV infection and will permit a reduction in the number of infected birds used in IBDV pathogenesis studies. The model can also be expanded to include other viruses and could be applied to different species of birds.

An Ex Vivo Chicken Primary Bursal-cell Culture Model to Study Infectious Bursal Disease Virus Pathogenesis

Here we describe the isolation of chicken primary bursal cells from the bursa of Fabricius, the culture and infection of the cells with infectious bursal disease virus, and the quantification of viral replication.

نموذج السابقين فيفو الابتدائي بورسال خلية ثقافة دجاج لدراسة الأمراض المعدية بورسال الفيروسات المرضية

Infectious bursal disease virus (IBDV) is a birnavirus of economic importance to the poultry industry. The virus infects B cells, causing morbidity, ...

A Murine Model of Dengue Virus-induced Acute Viral Encephalitis-like Disease

Dengue virus (DENV), an arthropod-borne virus transmitted by mosquitoes, may cause the severe disease known as dengue hemorrhagic fever, which is characterized by lethal complications due to plasma leakage, ascites, pleural effusion, respiratory distress, severe bleeding, and organ impairment. A few cases of DENV infection present neurological manifestations; however, studies have not explored DENV-induced neuropathogenesis further. In this study, we present a protocol to use an immunocompetent outbred ICR (Institute of Cancer Research) mouse for investigating the induction of central nervous system (CNS) infection with DENV, followed by the progression of acute viral encephalitis-like disease.

Here, we present a protocol for creating an immunocompetent ICR (Institute of Cancer Research) murine model of central nervous system infection to display the development of neuropathy. Monitoring acute viral encephalitic disorders by identical disease scores could be performed for showing dengue virus-induced neuropathy in vivo.

نموذج مورين من مرض التهاب الدماغ الفيروسي الحاد الناجم عن فيروس حمى الضنك

Dengue virus (DENV), an arthropod-borne virus transmitted by mosquitoes, may cause the severe disease known as dengue hemorrhagic fever, which is ...

Induction of Drug-Induced, Autoimmune Hepatitis in BALB/c Mice for the Study of Its Pathogenic Mechanisms

Drug-induced autoimmune hepatitis (DIH) is the most common hepatic drug-induced hypersensitization process observed in approximately 9 to 12% of patients with autoimmune hepatitis. The overwhelming majority of patients with DIH are women. The underlying mechanisms of these sex differences in prevalence are unclear because of the paucity of animal models that mimic human disease. Even so, underlying mechanisms are widely believed to be associated with human leukocyte antigen haplotypes and sex hormones. In contrast, using a DIH mouse model, we have uncovered that IL-4 initiated&#160;CD4+ T cells directed against an epitope of cytochrome P450 2E1 induces influx of neutrophils, macrophages and mast cells into the livers of female BALB/c mice. Using this model, we have also shown that IL-33-induced FoxP3+regulatory T cells confer protection against DIH in female and male mice. This DIH model is induced by immunizing mice with an epitope of CYP2E1 that has been covalently altered with a drug metabolite that has been associated with DIH. This epitope is recognized by patients with DIH. Our method induces robust and reproducible hepatitis and autoantibodies that can be utilized to study the pathogenesis of DIH. While in vivo studies can cause undue pain and distress in mice when done improperly, the advantage of an in vivo model is the ability to evaluate the pathogenesis of disease in a large number of mice. Additionally, biological effects of the altered liver proteins can be studied using invasive procedures. The addition of in vitro studies to the experimental design allows rapid repetition and mechanistic analysis at a cellular level. Thus, we will demonstrate our model protocol and how it can be utilized to study in vivo and in vitro mechanisms of DIH.

We describe an in vivo immunization, translational hepatitis model in BALB/c mice that can be utilized to study the pathogenesis of drug-induced autoimmune hepatitis including sex differences seen in this disease. We will describe how this model demonstrates reproducible analyses using in vivo and in vitro experimental techniques.

تحريض التهاب الكبد المناعي الذاتي الناجم عن الأدوية في الفئران BALB / c لدراسة آلياته المسببة للأمراض

Drug-induced autoimmune hepatitis (DIH) is the most common hepatic drug-induced hypersensitization process observed in approximately 9 to 12% of patients ...

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 ...

Propagation of the Microsporidian Parasite Edhazardia aedis in Aedes aegypti Mosquitoes

Edhazardia aedis is a microsporidian parasite of Aedes aegypti mosquitoes, a disease vector that transmits multiple arboviruses which cause millions of disease cases each year. E. aedis causes mortality and reduced reproductive fitness in the mosquito vector and has been explored for its potential as a biocontrol agent. The protocol we present for culturing E. aedis is based on its natural infection cycle, which involves both horizontal and vertical transmission at different life stages of the mosquito host. Ae. aegypti mosquitoes are exposed to spores in the larval stage. These infected larvae then mature into adults and transmit the parasite vertically to their offspring. Infected offspring are then used as a source of spores for future horizontal transmission. Culturing E. aedis can be challenging to the uninitiated given the complexities of the parasite&#8217;s life cycle, and this protocol provides detailed guidance and visual aids for clarification.

Propagation of the Microsporidian Parasite Edhazardia aedis in Aedes aegypti Mosquitoes

A protocol to culture the microsporidian parasite Edhazardia aedis. The parasite is passaged from one generation of Aedes aegypti mosquitoes to the next via horizontal transfer at the larval stage followed by vertical transmission at the adult stage. Live sporoplasms survive long-term in infected eggs.

انتشار الطفيلي Microsporidian Edhazardia aedis في Aedes aegypti البعوض

Edhazardia aedis is a microsporidian parasite of Aedes aegypti mosquitoes, a disease vector that transmits multiple arboviruses which cause millions of ...

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.