我々はナンシーR.シュトゥルム、免疫学、微生物学および分子生物学科、原稿読み取りのために重要な医学部のデビッドゲフィン学校、カリフォルニア大学ロサンゼルス校、世話になっています。研究·CNPqの全国協議会、および研究開発 - FAPDF、ブラジル、財団は、研究をサポートしていました。我々は、ブラジリア、ブラジルの大学から、アレッサンドロO.ソウザ、マリアC. Guimaro、シロコルデイロ、アナ·デ·カシアローザ、Roseneide·アウベス、ラファエル·アンドラーデの技術的な援助に感謝します。

<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. 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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>

利害の衝突が宣言されません。

生涯T.への影響を受けやすい哺乳類とは対照的に、 cruziの感染症は 、鶏がTに難治性であるクルーズトリパノソーマ感染 。鶏のモデルシステムの主な利点は、胎児の免疫系の開発において、初期感染の除去です。したがって、鶏の本体内に残っている唯一の寄生虫のDNAは、いくつかの遺伝子座で統合されています。 病原性Tの最適な量の使用クルーズは、肥沃な卵に接種しtrypomastigotesニワトリ胚のゲノムにkDNAのminicirclesの統合を取得に向けた重要なステップです。 100 trypomastigotesを接種した卵からのライブ雛の​​孵化率は500寄生虫で得られたものよりも4倍高くなっています。注意は卵の空気室に培養液の10μLに寄生虫懸濁液を接種するために注意する必要があります。卵白の漏れがあってはなりません。最適な条件下で、細胞内寄生虫感染が行わwiを取るその後感染が自然免疫によって排除されています。数時間一週間のための宿主細胞に進み内部インキュベーションと寄生虫乗算した後、薄い。 kDNAの統合は、生きている感染症を必要とし、初期胚の鶏の卵に裸minicirclesの接種は、統合にはなりません。 kDNA陽性胚およびコントロールは、37.5°Cと65％の湿度で制御された条件下で飼育する必要があります。雛は33℃の室温で2週間のケージに保管されています。その後、鶏が22℃の部屋で1.5メートル幅の通路で区切られた懸濁ラックにケージに保管され℃で濾過空気と動物福祉の条件を確保するために一定の疲労下の正圧である。大人の鶏焼きそば供給され、生後5ヶ月で卵を産む、完全な成長と成熟を達成するために飲用水道水を飲んでています。 T.を扱う際の衛生手順のメンテナンスは、結果の再現性に不可欠であるクルーズでは、接種肥沃な鶏の卵に。 T.鶏のモデルシステムのcruziの感染症は 、胚の成長の早い時期の免疫システムの開発の後に根絶されています。さらに、感染フリー、T.から孵化雛であることにクルーズでは、寄生虫抗原に対する寛容である、特定の抗体の欠如に、卵を接種した。細胞傷害性リンパ球（最小限の除去装置は、 図3）によって目標心拍細胞の拒絶反応は、免疫学的監視の3遺伝子型の変更や故障を示すkDNA変異ニワトリで見られます。遺伝子型変更されたT細胞は、体内で自己組織の拒絶反応の加速を示す。主な病変部位では、シャーガス病の特徴である心です。 （サーベイランス）生理学からphysiopathologic状態への通路は、細胞傷害性リンパ球3のクローン増殖を示すkDNA変異ニワトリで見られます。 T彼transkingdomモデルシステムは、T.によるゲノムの修飾に起因する、寄生虫誘発性、遺伝的に主導型自己免疫疾患（ 図6）を示します。 クルーズ kDNAは、統合をminicircle。これらの変更は、LB-接種卵から孵化した雛で見ていません。 この現象は自己組織の拒絶反応を防ぐために薬物の心筋に浸潤特異的T細胞の表現型の骨髄前駆細胞の抑制、組織適合性健康な骨髄の移植が必要な場合がありますkDNA変異鶏の炎症性自己免疫性心筋症の実験的治療法を示唆している。

<table border="1"><tbody><tr><td> 試薬の名前 </td><td> 会社 </td><td> カタログ番号 </td><td> コメント </td></tr><tr><td>組換えTaq DNAポリメラーゼ</td><td>インビトロジェン</td><td> 11615-010 </td><td></td></tr><tr><td>プラチナTaq DNAポリメラーゼ</td><td>インビトロジェン</td><td> 10966-030 </td><td></td></tr><tr><td>ランダムプライマーDNAラベリングシステム</td><td>インビトロジェン</td><td> 18187-013 </td><td></td></tr><tr><td> エコ RI </td><td>インビトロジェン</td><td> 15202-021 </td><td></td></tr><tr><td> MBO I </td><td>インビトロジェン</td><td> 15248-016 </td><td></td></tr><tr><td> dNTPのセット、100mMのソリューション</td><tD&gt; GEヘルスケア</td><td> 28-4065-51 </td><td></td></tr><tr><td>アマシャムハイボンド- N + -猫のn。 </td><td> GEヘルスケア</td><td> RPN303B </td><td></td></tr><tr><td> PlasmidPrepミニスピンキット</td><td> GEヘルスケア</td><td> 28-9042-70 </td><td></td></tr><tr><td> NSI I </td><td>シグマアルドリッチ</td><td> R5884 1,000個</td><td></td></tr><tr><td> DNA、ナトリウム塩魚精子</td><td> AMRESCO </td><td> 0644-10G </td><td></td></tr><tr><td>マウス抗鶏富栄-1B </td><td> SouthernBiotech </td><td> 8370から02 </td><td></td></tr><tr><td>マウス抗ニワトリCD45 </td><td> SouthernBiotech </td><td> 8270から08 </td><td></td></tr><tr><td>マウス抗ニワトリTCRγδ </td><td> SouthernBiotech </td><td> 8230から08 </td><td></td></tr><tr><td>マウス抗ニワトリCD8α </td><td> SouthernBiotech </td><td> 9220から02 </td><td></td></tr><tr><td>マウス抗ニワトリの単球/マクロファージ</td><td> SouthernBiotech </td><td> 8420から02 </td><td></td></tr><tr><td> MyCycle Termocycler </td><td>バイオ·ラッドラボラトリーズ</td><td> 580BR 5501 </td><td></td></tr></tbody></table>

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

乳児期および小児期に取得したクルーズトリパノソーマ急性感染症は無症候性に見えますが、慢性感染例の約三分の一が三十年以降にシャーガス病を示しています。自己免疫と寄生虫の永続性は、シャーガス病1、2の病態を説明する理論を競っています。シャーガス病の原虫の永続性と自己免疫が果たす役割を分離するために、我々はTを接種する受精卵の空気室でクルーズ 。成熟した鶏の免疫システムは Tに対してタイトな生物学的障壁であるクルーズと感染症は、成長3の最初の週の終わりまでに、その免疫システムの開発に根絶されています。雛は孵化時に寄生虫フリーですが、彼らは寄生虫ミトコンドリアのキネトプラストDNA（kDNA）がその子孫に転送され、そのゲノム内に統合されたminicircle保持します。ニワトリゲノムのkDNA minicircle統合のドキュメントは、ターグによって得られたetedプライムTAIL-PCR、サザンハイブリダイゼーション、クローニング、シークエンシング3、4。 kDNAは、統合をminicircle破裂転写と免疫系因子のオープンリーディングフレームは、ホスファターゼ（GTPアーゼ）、アデニル酸シクラーゼとホスホリラーゼ（PKC、NF-κBの活性化、PI-3K）は、細胞生理学、成長、分化、3、5に関連付けられている- 7、および他の遺伝子の機能。エフェクター細胞傷害性リンパ球による目標心拍繊維の除去に起因する重篤な心筋炎は、ヒトのシャーガス病に見られるものと類似した炎症性心筋症を示すkDNA変異ニワトリで見られます。特に、心不全および骨格筋の弱点は、1番染色体8のジストロフィン遺伝子のkDNAの破裂と大人の鶏に存在しています。同様のgenotipic変化はエフェクターCD45 +、CD8γδ+、CD8αリンパ球によって行わ組織の破壊に関連付けられています。したがって、この原虫感染症は、遺伝的に駆動される自己免疫疾患を誘発することができます。

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

の接種<em&gt;クルーズトリパノソーマ</em&gt;インキュベーションの前に肥沃な卵に寄生kDNAは胚細胞のゲノムに統合をminicircleレンダリングします。交配は子孫への変異の垂直転送を明らかにする。 kDNAは、いくつかの染色体にコード領域に統合し、鶏が炎症性自己免疫性心疾患で死亡しています。

寄生虫は鶏のモデルに遺伝的に駆動される自己免疫性シャーガス心臓病を誘発される

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 ビュー.  University of Brasilia. の接種クルーズトリパノソーマインキュベーションの前に肥沃な卵に寄生kDNAは胚細胞のゲノムに統合をminicircleレンダリングします。交配は子孫への変異の垂直転送を明らかにする。 kDNAは、いくつかの染色体にコード領域に統合し、鶏が炎症性自己免疫性心疾患で死亡しています。

ビデオ: 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.

SJLマウスにおける実験的自己免疫性下垂体炎の誘導

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

免疫・感染学

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.

ミエリンオリゴデンドロサイト糖タンパク質（MOG<sub&gt; 35〜55</sub&gt;）C57BL / 6マウスにおける実験的自己免疫性脳脊髄炎（EAE）誘発

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.

マウスおよび種々の組織における免疫細胞の疾患に依存する分布の評価における実験的自己免疫性脳脊髄炎の誘導

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.

男性と女性からのナイーブの仲間にアメリカトリパノソーマの性行

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.

伝染性ファブリキウス嚢病ウイルスの病原性を研究するためのEx Vivo鶏ファブリキウス嚢細胞培養モデル

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.

デングウイルス誘発急性ウイルス脳炎様疾患のマウスモデル

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.

BALB/cマウスにおける薬物誘発性自己免疫性肝炎の誘導による発症機序の研究

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.

グルコース-6-リン酸イソメラーゼ(G6PI)誘発RAマウスにおけるiNKT細胞の導入免疫療法

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.

アエデスアエジプティ蚊における微小胞体寄生虫・エダリア・アダイスの伝播

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.