אנו חייבים תודה ננסי ר &#39;שטורם, המחלקה לאימונולוגיה, למיקרוביולוגיה מולקולרית ביולוגיה, דיוויד גפן בית הספר לרפואה, אוניברסיטת קליפורניה בלוס אנג&#39;לס, על קריאה ביקורתית של כתב היד. המועצה הלאומית למחקר-CNPq, והקרן למחקר ופיתוח-FAPDF, ברזיל, נתמך במחקר. אנו מודים על הסיוע הטכני של אלסנדרו א &#39;סוזה, מריה ג Guimaro, סירו Cordeiro, אנה דה Cassia רוזה, אלבס Roseneide, ורפאל אנדראדה, מהאוניברסיטה של ​​ברזיליה, ברזיל.

<ol>
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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. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=NIK+and+IKKbeta+interdependence+in+NF-kappaB+signalling--flux+analysis+of+regulation+through+metabolites.">NIK and IKKbeta interdependence in NF-kappaB signalling--flux analysis of regulation through metabolites.</a> Biosystems. 99, 140-149 (2010).</li><li>Karakhanova, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=ERK/p38+MAP-kinases+and+PI3K+are+involved+in+the+differential+regulation+of+B7-H1+expression+in+DC+subsets.">ERK/p38 MAP-kinases and PI3K are involved in the differential regulation of B7-H1 expression in DC subsets.</a> Eur. J. Immunol. 40, 254-266 (2010).</li><li>Finsterer, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+heart+in+human+dystrophinopathies.">The heart in human dystrophinopathies.</a> Cardiology. 99, 1-19 (2003).</li><li>Nitz, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Heritable+integration+of+kDNA+minicircle+sequences+from+Trypanosoma+cruzi+into+the+avian+genome:+insights+into+human+Chagas+disease.">Heritable integration of kDNA minicircle sequences from Trypanosoma cruzi into the avian genome: insights into human Chagas disease.</a> Cell. 118, 175-186 (2004).</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=Kinetoplast+DNA+in+trypanosomid+flagellates.">Kinetoplast DNA in trypanosomid flagellates.</a> Int. 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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>

אין ניגוד עניינים הצהיר.

בניגוד ליונקים רגישים כל החיים ט &#39; זיהומים cruzi, תרנגולות הם מגיבים ט cruzi זיהום. היתרון העיקרי של מערכת מודל עוף היא חיסול הזיהום מוקדם בפיתוח של מערכת החיסון העוברי. לכן, ה-DNA הטפיל היחיד שנותר בתוך הגוף עוף משולב ב לוקוסים מספר. השימוש כמות אופטימלית של ט ארסית cruzi trypomastigotes לחסן ביצה פורייה היא צעד קריטי לקראת קבלת שילוב של minicircles kDNA לתוך הגנום עובר עוף. שיעור הבקיעה הגוזלים חי ביצים מחוסן עם 100 trypomastigotes הוא פי ארבעה גבוה יותר מזה שהושג עם 500 טפילים. יש להקפיד לחסן את ההשעיה טפילים μL 10 בינוני התרבות לתוך תא האוויר ביצה. לא אמורה להיות דליפה של חלבון ביצה. בתנאים אופטימליים, זיהום טפילי תאיים מתרחש Wiדקה כמה שעות לאחר כפל הדגירה ואת הטפיל בתוך התמורה תאים המארח למשך שבוע, לאחר מכן את הזיהום מסולק על ידי חסינות מולדת. שילוב kDNA דורש זיהום חיים, ואת הרכבה של minicircles עירומים לתוך ביצי תרנגולת הראשונים העובר אינו תוצאה של שילוב. KDNA חיובי העוברים והבקרות יש שוכנו בתנאים מבוקרים על 37.5 מעלות צלזיוס והלחות 65%. הגוזלים מוחזקים בכלובים במשך שבועיים ב 33 ° C טמפרטורת החדר. לאחר מכן את התרנגולות מוחזקות בכלובים תלויים על מתלים נפרדים ב -1.5 מטר רוחב המעברים בחדר על 22 מעלות צלזיוס עם אוויר מסונן לחץ חיובי תחת עייפות מתמדת כדי להבטיח את תנאי הרווחה של בעלי חיים. המבוגרים נמאס עוף האוכל ושותה מים זורמים ראויים לשתייה להשיג צמיחה בגרות מלאה, להטיל ביצים על חמישה חודשים של גיל. ותחזוקה של נהלים היגיינה חיוניים שחזור של תוצאות בעבודה עם ט &#39; cruzi מחוסןבתוך ביצי תרנגולת פוריים. במערכת עוף מודל T. זיהומים cruzi הושרשו לאחר פיתוח של מערכת החיסון בתקופה המוקדמת של צמיחה לעובר. בנוסף להיותו זיהום חינם, הגוזלים הבוקעים מן ט cruzi מחוסן ביצים, בחוסר של נוגדנים ספציפיים, סובלניים כדי אנטיגנים הטפיל. דחייה של לב תאים היעד ידי לימפוציטים ציטוטוקסיים (יחידת דחייה מינימלית, איור 3) היא לראות את התרנגולות kDNA-מוטציה המראים שינויים גנוטיפ פירוט של מעקב החיסונית 3. את שונה genotypically מתאי T להציג דחייה מואצת של רקמות עצמי בגוף. האתר הנגע העיקרי הוא הלב, שהוא סימן ההיכר של המחלה Chagas. המעבר מ פיזיולוגי (מעקב) למצב physiopathologic נתפסת עוף kDNA-מוטציה מראה clonally ריבוי לימפוציטים ציטוטוקסיים 3. Tהמודל שלו transkingdom המערכת מציגה טפיל-Induced, גנטית מונע מחלה אוטואימונית (איור 6), הנובע שינויים הגנום של ט &#39; cruzi kDNA minicircle ואינטגרציות. שינויים אלה לא נראים הגוזלים שבקעו מן הביצים LB-מחוסן. תופעה זו עולה כי הטיפול הניסיוני של קרדיומיופתיה אוטואימונית דלקתית kDNA למוצרי מוטציה תרנגולות עשויים לדרוש תרופות דיכוי של מח העצם של אבי פנוטיפ T-cell ספציפיים חדירה שריר הלב, וכן השתלת מח עצם בריא histocompatible כדי למנוע את דחיית רקמות עצמית.

<table border="1" style=";text-align:right;direction:rtl"><tbody><tr><td> שם מגיב </td><td> חברה </td><td> מספר קטלוגי </td><td> תגובות </td></tr><tr><td> Taq פולימראז ה-DNA רקומביננטי </td><td> Invitrogen </td><td> 11615-010 </td><td></td></tr><tr><td> פלטינום Taq DNA פולימראז </td><td> Invitrogen </td><td> 10966-030 </td><td></td></tr><tr><td> Random סימון primers DNA מערכת </td><td> Invitrogen </td><td> 18187-013 </td><td></td></tr><tr><td> Eco RI </td><td> Invitrogen </td><td> 15202-021 </td><td></td></tr><tr><td> MBO אני </td><td> Invitrogen </td><td> 15248-016 </td><td></td></tr><tr><td> סט dNTP, פתרונות 100mm </td><tד&gt; GE Healthcare </td><td> 28-4065-51 </td><td></td></tr><tr><td> Amersham Hybond - N + - חתול n. </td><td> GE Healthcare </td><td> RPN303B </td><td></td></tr><tr><td> PlasmidPrep מיני ספין ערכת </td><td> GE Healthcare </td><td> 28-9042-70 </td><td></td></tr><tr><td> NSI אני </td><td> Sigma-Aldrich </td><td> R5884 1KU </td><td></td></tr><tr><td> , ה-DNA מלח נתרן דגים זרע </td><td> AMRESCO </td><td> 0644-10G </td><td></td></tr><tr><td> נגד עוף עכבר bu-1B </td><td> SouthernBiotech </td><td> 8370-02 </td><td></td></tr><tr><td> עכבר נגד עוף CD45 </td><td> SouthernBiotech </td><td> 8270-08 </td><td></td></tr><tr><td>עכבר נגד עוף TCRγδ </td><td> SouthernBiotech </td><td> 8230-08 </td><td></td></tr><tr><td> עכבר נגד עוף CD8α </td><td> SouthernBiotech </td><td> 9220-02 </td><td></td></tr><tr><td> עכבר נגד עוף מונוציטים / מקרופגים </td><td> SouthernBiotech </td><td> 8420-02 </td><td></td></tr><tr><td> MyCycle Termocycler </td><td> Bio-Rad Laboratories </td><td> 580BR 5501 </td><td></td></tr></tbody></table>

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

את cruzi Trypanosoma זיהומים חריפים שנרכשו בינקות ובילדות נראה ללא תסמינים, אך כשליש מן המקרים שנדבקו במחלה כרונית להראות Chagas עד שלוש שנים או במאוחר. אוטואימוניות והתמדה טפיל מתחרים תיאוריות כדי להסביר את הפתוגנזה של המחלה Chagas 1, 2. לתפקידים שונים שיחק על ידי התמדה טפיל אוטואימוניות במחלה Chagas לנו לחסן ט cruzi בחדר אווירה של ביציות מופרות. המערכת החיסונית היא עוף בוגר מחסום ביולוגי חזק נגד ט &#39; cruzi ואת הזיהום הוא נמחק על התפתחות המערכת החיסונית שלה עד סוף השבוע הראשון של הצמיחה 3. האפרוחים הם טפיל ללא ב הבקיעה, אבל הם שומרים משולב טפיל המיטוכונדריה kinetoplast DNA (kDNA) minicircle בתוך הגנום שלהם המועברים לצאצאים. תיעוד של אינטגרציה kDNA minicircle בגנום עוף הושג על ידי טארגeted ראש זנב PCR, הכלאות הדרום, שיבוט, וכן רצף 3, 4. KDNA minicircle ואינטגרציות הקרע מסגרות קריאה פתוחות עבור שעתוק גורמים במערכת החיסון, phosphatase (GTPase), adenylate cyclase ו phosphorylases (PKC, NF-kappa B activator, PI-3K) הקשורים לפיזיולוגיה התא, צמיחה, והבחנה 3, 5 - 7, ופונקציות גנים אחרים. דלקת שריר הלב חריפה עקב דחייה של סיבי הלב היעד על ידי לימפוציטים ציטוטוקסיים effectors היא לראות את העופות שעברו מוטציה kDNA, מראה cardiomyopathy דלקתית דומה לזה ביטוי המחלה Chagas האדם. יש לציין כי אי ספיקת לב, חולשה בשרירי השלד נמצאים עופות בוגרים עם הקרע kDNA של גן הדיסטרופין על כרומוזום 1 8. שינויים דומים genotipic קשורים להרס רקמות שביצע effectors CD45 +, CD8γδ +, CD8α לימפוציטים. כך זה יכול לגרום זיהום protozoan מחלה אוטואימונית מונע גנטית.

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

חיסון של<em&gt; Trypanosoma cruzi</em&gt; ביצים פוריות שקדמו הדגירה הופך kDNA טפיל minicircle אינטגרציה בגנום העובר תאים. הכלאה מגלה העברה אנכית של המוטציות על הצאצאים. KDNA משתלב קידוד אזורי ב כרומוזומים מספר התרנגולות למות ממחלות לב דלקתית אוטואימונית.

טפיל מושרה מונע גנטית אוטואימונית מחלות לב Chagas דגם עוף

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

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Immunology and Infection

13.7K Views.  University of Brasilia. חיסון של Trypanosoma cruzi ביצים פוריות שקדמו הדגירה הופך kDNA טפיל minicircle אינטגרציה בגנום העובר תאים. הכלאה מגלה העברה אנכית של המוטציות על הצאצאים. KDNA משתלב קידוד אזורי ב כרומוזומים מספר התרנגולות למות ממחלות לב דלקתית אוטואימונית.

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.

אינדוקציה של Hypophysitis אוטואימוניות הניסויי עכברים 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.

המיאלין oligodendrocyte גליקופרוטאין (MOG<sub&gt; 35-55</sub&gt;) מושרה ניסויי אוטואימוניות Encephalomyelitis (EAE) בעכברים C57BL / 6

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.

אינדוקציה של ניסיוני אוטואימוניות Encephalomyelitis עכברים וההערכה של הפצת מחלות התלויה של תאים חיסוניים ברקמות שונות

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.

העברה מינית של Trypanosomes האמריקאי של זכרים ונקבות כדי החברים נאיבי

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.

מודל תרבות Bursal-תא ראשי לשעבר Vivo עוף ללמוד פתוגנזה וירוס מחלה זיהומית Bursal

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.

מודל Murine של וירוס דנגה-הנגרמת חריפה דלקת קרום המוח-כמו מחלה

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.

אינדוקציה של הפטיטיס אוטואימונית הנגרמת על ידי תרופות בעכברי BALB/c לחקר המנגנונים הפתוגניים שלה

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.

חיסוני המאמצת של תאים iNKT ב גלוקוז-6-פוספט איזומראז (G6PI)-הנגרמת עכברים 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.

התפשטות הטפיל המיקרו-ספורידיאן אדהאזרדיה אדיס ב-Aedes aegypti יתושים

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.