Name: chronic salmonella infection induced intestinal fibrosis
Uploaded: 2019-09-22T14:00:00
Description: Watch this Scientific Journal Video about Chronic Salmonella Infection Induced Intestinal Fibrosis at JoVE.com

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所有动物协议都得到不列颠哥伦比亚大学动物护理委员会的批准。
1. 为小鼠口服腹腔培养沙门氏菌的制备[AroA培养物]
<ol><li>从S的冷冻甘油库存。Typhimrium _AroA，使用含有100μg/mL链霉素的LB琼脂剂制备条纹板，具有无菌接种回路。在37°C下孵育过夜。条纹板可在 4°C 下存储长达一周。</li>
	<li>感染前一天，在2.5 mL水中溶解0.5克链霉素，制备抗生素。过滤灭菌链霉素?...

<ol>
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H., Powrie, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mouse+models+of+intestinal+inflammation+as+tools+to+understand+the+pathogenesis+of+inflammatory+bowel+disease.">Mouse models of intestinal inflammation as tools to understand the pathogenesis of inflammatory bowel disease.</a> European Journal of Immunology. 39 (8), 2021-2026 (2009).</li><li>Nell, S., Suerbaum, S., Josenhans, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+impact+of+the+microbiota+on+the+pathogenesis+of+IBD:+lessons+from+mouse+infection+models.">The impact of the microbiota on the pathogenesis of IBD: lessons from mouse infection models.</a> Nature Reviews Microbiology. 8 (8), 564-577 (2010).</li><li>Grassl, G. A., Finlay, B. B. <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+enteric+Salmonella+infections.">Pathogenesis of enteric Salmonella infections.</a> Current Opinion in Gastroenterology. 24 (1), 22-26 (2008).</li><li>Barthel, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pretreatment+of+mice+with+streptomycin+provides+a+Salmonella+enterica+serovar+Typhimurium+colitis+model+that+allows+analysis+of+both+pathogen+and+host.">Pretreatment of mice with streptomycin provides a Salmonella enterica serovar Typhimurium colitis model that allows analysis of both pathogen and host.</a> Infection and Immunity. 71 (5), 2839-2858 (2003).</li><li>Grassl, G. A., Valdez, Y., Bergstrom, K., Vallance, B. A., Finlay, B. 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B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Molecular+mechanisms+of+Salmonella+virulence+and+host+resistance.">Molecular mechanisms of Salmonella virulence and host resistance.</a> Current Topics in Microbiology and Immunology. 337, 93-127 (2009).</li><li>Valdez, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Nramp1+drives+an+accelerated+inflammatory+response+during+Salmonella-induced+colitis+in+mice.">Nramp1 drives an accelerated inflammatory response during Salmonella-induced colitis in mice.</a> Cellular Microbiology. 11 (2), 351-362 (2009).</li><li>Lo, B. C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+orphan+nuclear+receptor+RORalpha+and+group+3+innate+lymphoid+cells+drive+fibrosis+in+a+mouse+model+of+Crohn's+disease.">The orphan nuclear receptor RORalpha and group 3 innate lymphoid cells drive fibrosis in a mouse model of Crohn's disease.</a> Science Immunology. 1 (3), eaaf8864 (2016).</li><li>Burke, J. P., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fibrogenesis+in+Crohn's+disease.">Fibrogenesis in Crohn's disease.</a> American Journal of Gastroenterology. 102 (2), 439-448 (2007).</li><li>Hogan, B. L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Repair+and+regeneration+of+the+respiratory+system:+complexity,+plasticity,+and+mechanisms+of+lung+stem+cell+function.">Repair and regeneration of the respiratory system: complexity, plasticity, and mechanisms of lung stem cell function.</a> Cell Stem Cell. 15 (2), 123-138 (2014).</li><li>Fiocchi, C., Lund, P. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Themes+in+fibrosis+and+gastrointestinal+inflammation.">Themes in fibrosis and gastrointestinal inflammation.</a> American Journal of Physiology-Gastrointestinal and Liver Physiology. 300 (5), G677-G683 (2011).</li><li>Rieder, F., Fiocchi, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intestinal+fibrosis+in+IBD&#8212;a+dynamic,+multifactorial+process.">Intestinal fibrosis in IBD&#8212;a dynamic, multifactorial process.</a> Nature Reviews Gastroenterology &amp; Hepatology. 6 (4), 228-235 (2009).</li><li>Bouguen, G., Peyrin-Biroulet, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Surgery+for+adult+Crohn's+disease:+what+is+the+actual+risk?.">Surgery for adult Crohn's disease: what is the actual risk?.</a> Gut. 60 (9), 1178-1181 (2011).</li><li>Wynn, T. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cellular+and+molecular+mechanisms+of+fibrosis.">Cellular and molecular mechanisms of fibrosis.</a> The Journal of Pathology. 214 (2), 199-210 (2008).</li><li>Junqueira, L. C., Bignolas, G., Brentani, R. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Picrosirius+staining+plus+polarization+microscopy,+a+specific+method+for+collagen+detection+in+tissue+sections.">Picrosirius staining plus polarization microscopy, a specific method for collagen detection in tissue sections.</a> Histochem J. 11 (4), 447-455 (1979).</li><li>Lo, B. C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=IL-22+Preserves+Gut+Epithelial+Integrity+and+Promotes+Disease+Remission+during+Chronic+Salmonella+Infection.">IL-22 Preserves Gut Epithelial Integrity and Promotes Disease Remission during Chronic Salmonella Infection.</a> Journal of Immunology. 202 (3), 956-965 (2019).</li><li>Fichtner-Feigl, S., et al. <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+IL-13+triggers+TGF-beta1-dependent+tissue+fibrosis+in+chronic+2,4,6-trinitrobenzene+sulfonic+acid+colitis.">Induction of IL-13 triggers TGF-beta1-dependent tissue fibrosis in chronic 2,4,6-trinitrobenzene sulfonic acid colitis.</a> Journal of Immunology. 178 (9), 5859-5870 (2007).</li><li>Fichtner-Feigl, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=IL-13+signaling+via+IL-13R+alpha2+induces+major+downstream+fibrogenic+factors+mediating+fibrosis+in+chronic+TNBS+colitis.">IL-13 signaling via IL-13R alpha2 induces major downstream fibrogenic factors mediating fibrosis in chronic TNBS colitis.</a> Gastroenterology. 135 (6), e2001-e2007 (2013).</li><li>Johnson, L. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intestinal+fibrosis+is+reduced+by+early+elimination+of+inflammation+in+a+mouse+model+of+IBD:+impact+of+a+&amp;#34;Top-Down&amp;#34;+approach+to+intestinal+fibrosis+in+mice.">Intestinal fibrosis is reduced by early elimination of inflammation in a mouse model of IBD: impact of a &amp;#34;Top-Down&amp;#34; approach to intestinal fibrosis in mice.</a> Inflammatory Bowel Diseases. 18 (3), 460-471 (2012).</li><li>Darfeuille-Michaud, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=High+prevalence+of+adherent-invasive+Escherichia+coli+associated+with+ileal+mucosa+in+Crohn's+disease.">High prevalence of adherent-invasive Escherichia coli associated with ileal mucosa in Crohn's disease.</a> Gastroenterology. 127 (2), 412-421 (2004).</li><li>Small, C. L., Reid-Yu, S. A., McPhee, J. B., Coombes, B. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Persistent+infection+with+Crohn's+disease-associated+adherent-invasive+Escherichia+coli+leads+to+chronic+inflammation+and+intestinal+fibrosis.">Persistent infection with Crohn's disease-associated adherent-invasive Escherichia coli leads to chronic inflammation and intestinal fibrosis.</a> Nature Communications. 4, 1957 (2013).</li><li>Imai, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Flagellin-mediated+activation+of+IL-33-ST2+signaling+by+a+pathobiont+promotes+intestinal+fibrosis.">Flagellin-mediated activation of IL-33-ST2 signaling by a pathobiont promotes intestinal fibrosis.</a> Mucosal Immunology. , (2019).</li></ol>

我们对IBD发病机制的理解已经大大增强了小鼠的肠道炎症模型。虽然这些个体模型并不概括复杂和多因素人类疾病的所有特征，但它们有助于确定疾病进展的关键特征。与IBD相关的纤维化严格仍然是一个主要未满足的临床需要，因为目前的治疗在逆转疾病发展方面是无效的。此外，由于目前动物模型的限制，肠纤维化在实验室环境中很难研究。BALB/c小鼠长期接触TNBS已被证明诱导由IL-13和TGF-B1<sup cl...

溃疡性结肠炎（UC）和克罗恩病（CD）是IBD的两种主要形式，并被定性为胃肠道1、2的慢性和复发性炎症性疾病。这些疾病对患者的生活质量有重大影响。IBD的症状包括腹痛，腹泻，恶心，体重减轻，发烧和疲劳3。最近的研究已经确定了导致疾病发病机制的遗传和环境因素;据认为，这些危险因素导致上皮屏障的中断，导致发光抗原4的易位或过采样。因此，这启动一个异常的炎症反应，由肠道免疫细胞4介导的共体菌群。IBD相关并发症的特征可能延伸到胃肠道以外的部位，影响各种器官，包括关节、皮肤和肝脏1，2。UC的标记包括严重和弥漫性炎症，通常局部在结肠1。疾病病理学影响肠道的粘膜和亚粘膜，导致表面粘膜溃疡1。相反，CD可以影响胃肠道的任何部分，虽然疾病的证据常见于结肠和远端回肠2。此外，CD中的炎症是跨体，影响肠壁的所有层2。
已识别的几个IBD易感基因将表明上皮屏障或免疫调节失调是疾病进展5的关键贡献者。发现单核细胞表达的核苷酸寡聚域2（NOD2）中的突变与CD易感性增加有关;这突出了细菌成分的先天免疫检测与疾病6之间的联系。最近的全基因组关联研究（GWAS）揭示了其他可能参与IBD发病机制的通路，包括：STAT1、NKX2-3、IL2RA、IL23R依赖途径的遗传变异与适应性免疫、MUC1、MUC19和PTGER4在肠道屏障维护中，以及ATG16L介导自噬7，8，9。虽然这些基于人群的遗传学研究增强了我们对IBD的理解，但仅仅易感等位基因可能不足以引发和维持慢性疾病3。其他非遗传因素，包括肠道微生物群的改变和多样性的减少都与肠道炎症有关。然而，目前还不清楚肠道消化不良是先于或是否是免疫反应不良的后果3。虽然IBD的病因尚不明确，但通过10、11的实验性小鼠模型，我们加强了对疾病发病机制的了解。这些模型单独并不完全代表人类疾病的复杂性，但它们对于阐明与IBD相关的病理生理学途径和验证试探性治疗策略10具有价值。 11.这种小鼠模型通常依赖于通过化学诱导或感染、免疫细胞转移或基因操作引发炎症。此外，这些策略往往涉及上皮完整性的扰动或先天或适应性免疫的调制。
肠沙门氏菌是肠道病原体，可以感染人类和小鼠。摄入后，沙门氏菌可以通过直接侵入上皮细胞、M细胞或抗原呈现细胞12来殖民肠道。被S口头感染的老鼠。Typhimurium导致主要在系统位点的殖民化，如脾脏淋巴结，在GI区12中丰度相对较低。然而，对使用链霉素的小鼠进行预处理，通过减少正常微生物群13的宿主保护作用，提高了肠道沙门氏菌的殖民化效率。该模型的病理特征包括上皮屏障的中断或溃疡、粒细胞招募和严重水肿13。或者，感染S级疫苗。Typhimurium _AroA 突变体导致慢性殖民化的 cecum 和结肠，持续到感染14后的第 40 天。S.Typhimurium_AroA菌株在芳香氨基酸的生物合成中存在缺陷;这使得突变菌株具有阿维性，并可用作高效疫苗15。小鼠的口腔感染导致Th1-和Th17细胞因子相关的炎症反应、广泛的组织重塑和胶原蛋白沉积。组织病理学与亲纤维化因子（如TGF-β1、CTGF和IGF14）的升高有关。该模型中报告的跨壁画纤维化疤痕让人联想到IBD中经常观察到的严格性形成。沙门氏菌引起的纤维化需要由沙门氏菌致病性岛屿（SPI）-1和2 12编码的毒性。重要的是，这个S.Tymphimurium_AroA感染模型是研究在C57/Bl6背景下维持的突变小鼠纤维化反应的有用系统。C57/Bl6 应变对 S 极其敏感。由于基因编码自然抗药性相关巨噬平板蛋白（NRAMP）-116、17的功能丧失而感染。我们发现IL-17A和ROR®依赖性先天淋巴细胞是本模型18中发病机制的重要贡献者。
CD的一个主要并发症是细胞外基质（ECM）的调节失调和过度沉积，包括胶原蛋白2，19。虽然胃肠道具有相对较高的再生能力，纤维化疤痕可能会出现由于未解决的伤口愈合反应，与慢性和严重炎症20，21。在CD中，这导致对组织结构的有害影响，导致严重的器官损伤21，22。在CD中观察到的炎症的跨面性质最终先于肠壁的增厚与症状性狭窄或严格形成21相关。约三分之一的CD患者需要肠道切除这种并发症22。IBD没有有效的抗纤维化疗法，因为使用免疫抑制剂，如阿扎西普林或抗TNF®生物制剂没有影响，或只是适度减少手术干预的需求19，23.虽然纤维化被认为是慢性炎症的结果，但中质细胞（如成纤维细胞和围细胞）被认为是纤维化疤痕21、24中ECM的主要细胞来源。慢性S.Typhimurium _AroA 感染是肠道纤维化的一种强大的小鼠模型，可提供对 CD 样特征的发病机制的见解。

<table>
	<tbody>
		<tr>
			<td>2 ml round bottom safe lock tubes</td>
			<td>Eppendorf</td>
			<td>22363344</td>
			<td></td>
		</tr>
		<tr>
			<td>Stainless steel beads</td>
			<td>Qiagen</td>
			<td>69989</td>
			<td></td>
		</tr>
		<tr>
			<td>PBS</td>
			<td>Gibco</td>
			<td>10010031</td>
			<td></td>
		</tr>
		<tr>
			<td>Large-Orifice Pipet Tips</td>
			<td>Fisher</td>
			<td>2707134</td>
			<td></td>
		</tr>
		<tr>
			<td>2 mL megablock plates</td>
			<td>Sarstedt</td>
			<td>82.1972.002</td>
			<td></td>
		</tr>
		<tr>
			<td>Gavage needles</td>
			<td>FST</td>
			<td>18061-22</td>
			<td></td>
		</tr>
		<tr>
			<td>Streptomycin sulfate</td>
			<td>Sigma</td>
			<td>S9137</td>
			<td></td>
		</tr>
		<tr>
			<td>Mixer mill</td>
			<td>Retsch</td>
			<td>MM</td>
			<td></td>
		</tr>
	</tbody>
</table>

chronic salmonella infection induced intestinal fibrosis

组织纤维化的特点是细胞外基质（如胶原蛋白）的病理积累是持续炎症和失调修复的结果。在炎症性肠病（IBD）中，纤维化导致复发性严格性形成，除了手术切除，没有其他有效的治疗方法。由于其发病较晚，驱动纤维化的过程较少研究，而且在很大程度上是未知的。因此，纤维化并发症是IBD的一大挑战。在本议定书中，描述了肠道纤维化的强健体内模型，其中对C57Bl/6小鼠进行链霉素预处理，然后口服带有疫苗级沙门氏菌的口角，即阿罗阿突变体导致持久性病原体殖民化和纤维化。准备S.解释用于接种、定量囊肿和脾脏中的病原体负荷以及评估肠道组织中胶原蛋白沉积的Typhimurium_AroA。这种实验性疾病模型可用于检查增强或加剧CD样肠道纤维化的宿主因素。

链霉素治疗后，口服感染S.Typhimurium _AroA 导致强健的肠道炎症和纤维化，特别是在 cecum 中（图 1）。典型的病原体负担为每1克1克1至109 CFU，每1克脾脏可获104 CFU，可从受感染动物中恢复（图2）。在皮罗西里乌斯红染色剖腹产的纤维化评估表明，感染后21天出现纤维化，而大部分病理在第42 pi（图3和<s...

Watch this Scientific Journal Video about Chronic Salmonella Infection Induced Intestinal Fibrosis at JoVE.com

Chronic Salmonella Infection Induced Intestinal Fibrosis

该协议描述了沙门氏菌驱动的肠道纤维化的小鼠模型，类似于克罗恩病的关键病理特征，包括跨体炎症和纤维化。该方法可用于评估使用维持在C57Bl/6遗传背景的突变小鼠改变纤维化结果的宿主因素。

慢性沙门氏菌感染诱发肠纤维化

Tissue fibrosis characterized by the pathological accumulation of extracellular matrix such as collagen is the outcome of persistent inflammation and ...

组织纤维化是器官随着时间的推移而失效的主要方式，以响应慢性炎症或衰老。事实上，我认为，如果你不死于传染性病原体或癌症，你很可能会死于器官纤维化，随着年龄的增长。不幸的是，没有很多伟大的小鼠模型来模仿人体组织纤维化。因此，我们开发的是克罗恩病的小鼠模型，这是一种人类疾病，导致肠道纤维化，基本上无法治疗。事实上，唯一的治疗方式是通过手术切除纤维组织和肠的重新连接。我们模型的优点是，它在所有小鼠菌株中完全具有素化性，并且非常密切地模仿人类克罗恩病，纤维化持续一个多月，这真是一个健壮的模型。这个程序中最具挑战性的部分是小鼠的处理，特别是口腔的格夫，因为这需要大量的实践特殊认证。首次执行此程序时，应注意使用沙门氏菌需要适当的安全预防措施和无菌技术。在实验之前，应计划对废物和细菌进行处理。这个程序的视觉演示是非常重要的，因为它涉及到沙门氏菌的处理，需要特殊的安全预防措施，以及适当的无菌技术。为了开始这个程序，请用每毫升100微克的浓度设置一个含有链霉素的LB汞板。使用无菌接种回路，用S型三角箭头的冷冻甘油库存条纹板。在37摄氏度下孵育过夜。条纹板可以在四摄氏度下储存一周。感染前一天，在2.5毫升水中溶解0.5克链霉素，准备抗生素。过滤器对链霉素溶液进行消毒。然后，使用一个灯泡尖的22量表加夫针与一毫升注射器口头打磨小鼠与100微升链霉素溶液。接下来，将含有链霉素的三毫升 LB 肉汤加入培养管，浓度为每毫升 50 微克。使用接种循环，用单个菌落为LB汤接种。在37摄氏度下在37摄氏度下进行有氧孵育，在200转/小时时摇晃。在感染当天，连续两次使用无菌PBS稀释隔夜沙门氏菌培养物，以准备最终感染剂量。这导致一个100微升的内分，包含大约300万个菌落形成单位。在此之后，使用灯泡尖的22量量格维奇针与毫升注射器，用100微升的准备好沙门氏菌每只鼠标。首先，设置两毫升安全锁圆底微管，添加一毫升无菌PBS和一个自动 <3> <3>开锁的不锈钢珠到每个管。在组织收集之前预称管。对小鼠实施安乐死后，重新校正其脑电和脑组织，确保从单个动物身上收集组织，放入单独的管子中。称重每个管以确定组织重量。使用搅拌机设备以 30 赫兹使组织均质 15 分钟。然后，将 900 微升 PBS 转移到 96 井两毫升巨型块的每个井中。移液器100微升的组织均匀化成第一个井，并混合良好。通过向后续孔添加 100 微升来进行串行稀释，直到获得 10 到负的第六次稀释。板 10 微升每次稀释三钙到含有链霉素的 LB agar 上。然后，计算平均菌落形成单位，并确定每克组织形成的菌落单位，如文本协议中所述。打开斐济，这是一个开源的成像软件，并拖放到工具栏上的tif图像文件。在菜单栏上，选择"图像、类型"和"RGB"堆栈，将图像拆分为红色、绿色和蓝色通道。滑动面板底部的水平条，将通道设置为绿色。打开图像，调整，阈值工具。调整最小和最大限制以消除任何背景信号。设置所需阈值后，关闭阈值工具，然后转到"分析"、设置测量、检查"面积、面积分数、阈值限制"和"显示"标签。使用徒手选择或多边形选择工具获取组织选择，并单击"分析"，"测量"来测量胶原蛋白的正百分比区域。然后，将胶原蛋白染色的绝对区域正化为组织区域。链霉素治疗，然后口服感染S酪虫三角洲箭头导致强健的肠道炎症和纤维化，特别是在塞库姆。典型的病原体负担为1亿至10亿个菌落形成单位，可从受感染的动物身上每克聚居地中回收一万个菌群形成单位，而每克脾脏通常可回收1万个菌落形成单位。对皮罗西里乌斯红色染色的脑油部分的纤维化评估表明感染后21天出现纤维化高峰，而大部分病理学在感染后第42天就解决了。材料的彻底准备至关重要，因为这个实验需要多天时间进行。LB板和沙门氏菌和链霉素的制备都应该无菌完成。在实验之前，确保实验者熟悉动物协议以及安全隐患。在评估胶原蛋白负担时，实验者可以选择进行其他生物测定，如RNA测序，以评估基因表达特征或流动细胞学以评估免疫细胞子集或组织ELISA来评估细胞因子图谱。因此，我们的模型特别擅长模仿一种叫做克罗恩病的人类疾病，在那里你得到肠道纤维化，这在很大程度上是无法治疗的，唯一的治疗方式是去除肠道的一部分，并缝合在一起，希望它不会回来。我们的模型非常健壮，我们已经表明，针对先天淋巴细胞类型三，一个称为原始α的转录因子，或白细胞间17可刺激疾病，并防止纤维化。我们的模型特别善于识别克罗恩病和肠道纤维化发病机制中起作用的因素，我们已经在确定一些靶点的道路上，这些靶点可以通过治疗来缓解炎症性肠病。我们的模型已经进入相同的ILC3或α NIL17访问可能在其他纤维病发挥作用，其中包括像银屑病，多发性硬化症，特发性肺纤维化的东西。因此，针对这些相同的因素可能减轻这些疾病的纤维化。虽然突变沙门氏菌流不是致命的，但必须遵守工厂给出的标准生物危害安全协议。此外，由于甲醛蒸汽已知具有致癌性，因此在固定步骤中，在烟气罩后面工作非常重要。

Research

JoVE Journal

Immunology and Infection

Long Term Chronic Pseudomonas aeruginosa Airway Infection in Mice

Long Term Chronic Pseudomonas aeruginosa Airway Infection in Mice

长期慢性<em&gt;铜绿假单胞菌</em&gt;在呼吸道感染小鼠

A mouse model of chronic airway infection is a key asset in cystic fibrosis (CF) research, although there are a number of concerns regarding the model ...

科研

免疫与感染

A Mouse Model for Pathogen-induced Chronic Inflammation at Local and Systemic Sites

鼠标型号病原体引起的慢性炎症局部及全身站点

Chronic inflammation is a major driver of pathological tissue damage and a unifying characteristic of many chronic diseases in humans including ...

A Protocol to Infect Caenorhabditis elegans with Salmonella typhimurium

A Protocol to Infect Caenorhabditis elegans with Salmonella typhimurium

一个协议感染<em&gt;秀丽隐杆线虫</em&gt;与<em&gt;鼠伤寒沙门氏菌</em

In the last decade, C. elegans has emerged as an invertebrate organism to study interactions between hosts and pathogens, including the host defense ...

Quantification of Cytosolic vs. Vacuolar Salmonella in Primary Macrophages by Differential Permeabilization

Quantification of Cytosolic vs. Vacuolar Salmonella in Primary Macrophages by Differential Permeabilization

胞浆的定量与液泡<em&gt;沙门氏菌</em&gt;在原发性巨噬细胞差透

Intracellular bacterial pathogens can replicate in the cytosol or in specialized pathogen-containing vacuoles (PCVs). To reach the cytosol, bacteria like ...

A Multimodal Imaging Approach Based on Micro-CT and Fluorescence Molecular Tomography for Longitudinal Assessment of Bleomycin-Induced Lung Fibrosis in Mice

基于显微 CT 和荧光分子层析成像的多模态成像方法对博莱霉素诱导的小鼠肺纤维化的纵向评价

Idiopathic pulmonary fibrosis (IPF) is a fatal lung disease characterized by the progressive and irreversible destruction of lung architecture, which ...

Isolation of Salmonella typhimurium-containing Phagosomes from Macrophages

Isolation of Salmonella typhimurium-containing Phagosomes from Macrophages

巨噬细胞中含有吞噬的鼠伤寒沙门氏菌的分离

Salmonella typhimurium is a facultative intracellular bacterium that causes gastroenteritis in humans. After invasion of the lamina propria, S. ...

Using Human Induced Pluripotent Stem Cell-derived Intestinal Organoids to Study and Modify Epithelial Cell Protection Against Salmonella and Other Pathogens

Using Human Induced Pluripotent Stem Cell-derived Intestinal Organoids to Study and Modify Epithelial Cell Protection Against Salmonella and Other Pathogens

利用人类诱导的多能干细胞衍生的肠道有机体研究和改性上皮细胞对沙门氏菌和其他病原体的保护

The intestinal &#8216;organoid&#8217; (iHO) system, wherein 3-D structures representative of the epithelial lining of the human gut can be produced from ...

Mechanistic Insight into the Development of TNBS-Mediated Intestinal Fibrosis and Evaluating the Inhibitory Effects of Rapamycin

对TNBS介导肠纤维化发展的力学洞察与拉帕霉素抑制作用的评价

Significant studies have been carried out to understand effective management of intestinal fibrosis. However, the lack of better knowledge of fibrosis has ...

A Delayed Inoculation Model of Chronic Pseudomonas aeruginosa Wound Infection

A Delayed Inoculation Model of Chronic Pseudomonas aeruginosa Wound Infection

慢性假单位病伤口感染的延迟接种模型

Pseudomonas aeruginosa (P. aeruginosa) is a major nosocomial pathogen of increasing relevance to human health and disease, particularly in the setting of ...

Systematic Scoring Analysis for Intestinal Inflammation in a Murine Dextran Sodium Sulfate-Induced Colitis Model

穆林·德克斯特兰硫酸钠诱导结肠炎模型中肠道炎症的系统评分分析

Murine colitis models are tools that are extensively employed in studies focused on understanding the pathobiology of inflammatory intestinal disorders. ...