Name: induction of drug-induced, autoimmune hepatitis in balb/c mice for the study of its pathogenic mechanisms
Uploaded: 2020-05-29T12:30:00
Description: Watch this Scientific Journal Video about Induction of Drug-Induced, Autoimmune Hepatitis in BALB/c Mice for the Study of Its Pathogenic Mechanisms at JoVE.com

Njoku博士要感谢医学博士Noel R. Rose博士的指导和富有洞察力的讨论，这些指导和讨论导致了该模型的制定。

所有程序均由动物护理和使用委员会批准。
1. 肝 S-100 胞质蛋白或 CYP2E1 表位的三氟乙酰化
注意：首先，制备三氟乙酰化S100（TFA-S100）和三氟乙酰基化CYP2E1表位（TFA-JHDN5）。因为免疫需要同源S100蛋白，并且需要BALB / c小鼠来产生免疫原。该制剂产生大量的免疫原;因此，预计每年执行此部分大约四次。将使用相同的方法来制造TFA-JHDN5。环氧乙烷2E1表位（JHDN5?...

<ol>
	<li>Castiella, A., Zapata, E., Lucena, M. I., Andrade, R. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Drug-induced+autoimmune+liver+disease:+A+diagnostic+dilemma+of+an+increasingly+reported+disease.">Drug-induced autoimmune liver disease: A diagnostic dilemma of an increasingly reported disease.</a> World J. Hepatology. 6 (4), 160-168 (2014).</li><li>Bjornsson, E. S., Bergmann, O. M., Bjornsson, H. K., Kvaran, R. B., Olafsson, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Incidence,+presentation,+and+outcomes+in+patients+with+drug-induced+liver+injury+in+the+general+population+of+Iceland.">Incidence, presentation, and outcomes in patients with drug-induced liver injury in the general population of Iceland.</a> Gastroenterology. 144 (7), 1419-1425 (2013).</li><li>Castiella, A., Lucena, M. I., Zapata, E. M., Otazua, P., Andrade, R. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Drug-induced+autoimmune-like+hepatitis:+a+diagnostic+challenge.">Drug-induced autoimmune-like hepatitis: a diagnostic challenge.</a> Digestive Diseases and Sciences. 56 (8), 2501-2502 (2011).</li><li>Czaja, A. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Drug-induced+autoimmune-like+hepatitis.">Drug-induced autoimmune-like hepatitis.</a> Digestive Diseases and Sciences. 56 (4), 958-976 (2011).</li><li>Pohl, L. R., Thomassen, D., Pumford, N. R., Butler, L. E., Satoh, H., Ferrans, V. J., Perrone, 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=Hapten+carrier+conjugates+associated+with+halothane+hepatitis.">Hapten carrier conjugates associated with halothane hepatitis.</a> Advances in Experimental Medicine and Biology. 283, 111-120 (1991).</li><li>Njoku, D. B., Talor, M. V., Fairweather, D., Frisancho-Kiss, S., Odumade, O. A., Rose, N. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+novel+model+of+drug+hapten-induced+hepatitis+with+increased+mast+cells+in+the+BALB/c+mouse.">A novel model of drug hapten-induced hepatitis with increased mast cells in the BALB/c mouse.</a> Experimental and Molecular Pathology. 78 (2), 87-100 (2005).</li><li>Cottagiri, M., Nyandjo, M., Stephens, M., Mantilla, J., Saito, H., Mackay, I. R., 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=In+drug-induced,+immune-mediated+hepatitis,+interleukin-33+reduces+hepatitis+and+improves+survival+independently+and+as+a+consequence+of+FoxP3++T-cell+activity.">In drug-induced, immune-mediated hepatitis, interleukin-33 reduces hepatitis and improves survival independently and as a consequence of FoxP3+ T-cell activity.</a> Cellular and Molecular Immunology. , (2018).</li><li>Lohse, A. W., Manns, M., Dienes, H. P., Meyer zum Buschenfelde, K. H., Cohen, I. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Experimental+autoimmune+hepatitis:+disease+induction,+time+course+and+T-cell+reactivity.">Experimental autoimmune hepatitis: disease induction, time course and T-cell reactivity.</a> Hepatology. 11 (1), 24-30 (1991).</li><li>McCarthy, E. K., Vakos, A., Cottagiri, M., Mantilla, J. J., Santhanam, L., Thomas, D. 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=Identification+of+a+Shared+Cytochrome+p4502E1+Epitope+Found+in+Anesthetic+Drug-Induced+and+Viral+Hepatitis.">Identification of a Shared Cytochrome p4502E1 Epitope Found in Anesthetic Drug-Induced and Viral Hepatitis.</a> mSphere. 3 (5), (2018).</li><li>Cho, J., Kim, L., Li, Z., Rose, N. R., Talor, M. V., Njoku, D. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sex+bias+in+experimental+immune-mediated,+drug-induced+liver+injury+in+BALB/c+mice:+suggested+roles+for+Tregs,+estrogen,+and+IL-6.">Sex bias in experimental immune-mediated, drug-induced liver injury in BALB/c mice: suggested roles for Tregs, estrogen, and IL-6.</a> PLoS. One. 8 (4), 61186 (2013).</li><li>Satoh, H., Gillette, J. R., Takemura, T., Ferrans, V. J., Jelenich, S. E., Kenna, J. G., 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=Investigation+of+the+immunological+basis+of+halothane-induced+hepatotoxicity.">Investigation of the immunological basis of halothane-induced hepatotoxicity.</a> Advances in Experimental Medicine and Biology. 197, 657-673 (1986).</li><li>Eliasson, E., Kenna, J. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cytochrome+P450+2E1+is+a+cell+surface+autoantigen+in+halothane+hepatitis.">Cytochrome P450 2E1 is a cell surface autoantigen in halothane hepatitis.</a> Molecular Pharmacology. 50 (3), 573-582 (1996).</li><li>Bourdi, M., Chen, W., Peter, R. M., Martin, J. L., Buters, J. T., Nelson, S. D., 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=Human+cytochrome+P450+2E1+is+a+major+autoantigen+associated+with+halothane+hepatitis.">Human cytochrome P450 2E1 is a major autoantigen associated with halothane hepatitis.</a> Chemical Research in Toxicology. 9 (7), 1159-1166 (1996).</li><li>Njoku, D. B., Li, Z., Washington, N. D., Mellerson, J. L., Talor, M. V., Sharma, R., 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=Suppressive+and+pro-inflammatory+roles+for+IL-4+in+the+pathogenesis+of+experimental+drug-induced+liver+injury.">Suppressive and pro-inflammatory roles for IL-4 in the pathogenesis of experimental drug-induced liver injury.</a> European Journal of Immunology. 39 (6), 1652-1663 (2009).</li><li>Aithal, G. P., Ramsay, L., Daly, A. K., Sonchit, N., Leathart, J. B., Alexander, G., 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=Hepatic+adducts,+circulating+antibodies,+and+cytokine+polymorphisms+in+patients+with+diclofenac+hepatotoxicity.">Hepatic adducts, circulating antibodies, and cytokine polymorphisms in patients with diclofenac hepatotoxicity.</a> Hepatology. 39 (5), 1430-1440 (2004).</li><li>Higuchi, S., Kobayashi, M., Yoshikawa, Y., Tsuneyama, K., Fukami, T., Nakajima, 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=IL-4+mediates+dicloxacillin-induced+liver+injury+in+mice.">IL-4 mediates dicloxacillin-induced liver injury in mice.</a> Toxicology Letters. 200 (3), 139-145 (2011).</li><li>Rubtsova, K., Marrack, P., Rubtsov, A. V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sexual+dimorphism+in+autoimmunity.">Sexual dimorphism in autoimmunity.</a> Journal of Clinical Investigation. 125 (6), 2187-2193 (2015).</li><li>Satoh, H., Fukuda, Y., Anderson, D. K., Ferrans, V. J., Gillette, J. R., Pohl, L. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Immunological+studies+on+the+mechanism+of+halothane-induced+hepatotoxicity:+immunohistochemical+evidence+of+trifluoroacetylated+hepatocytes.">Immunological studies on the mechanism of halothane-induced hepatotoxicity: immunohistochemical evidence of trifluoroacetylated hepatocytes.</a> Journal of Pharmacology and Experimental Therapeutics. 233 (3), 857-862 (1985).</li><li>Habeeb, A. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Determination+of+free+amino+groups+in+proteins+by+trinitrobenzenesulfonic+acid.">Determination of free amino groups in proteins by trinitrobenzenesulfonic acid.</a> Analytical Biochemistry. 14 (3), 328-336 (1966).</li><li>Christen, U., Burgin, M., Gut, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Halothane+metabolism:+immunochemical+evidence+for+molecular+mimicry+of+trifluoroacetylated+liver+protein+adducts+by+constitutive+polypeptides.">Halothane metabolism: immunochemical evidence for molecular mimicry of trifluoroacetylated liver protein adducts by constitutive polypeptides.</a> Molecular Pharmacology. 40 (3), 390-400 (1991).</li><li>Christen, U., Quinn, J., Yeaman, S. J., Kenna, J. G., Clarke, J. B., Gandolfi, A. 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=Identification+of+the+dihydrolipoamide+acetyltransferase+subunit+of+the+human+pyruvate+dehydrogenase+complex+as+an+autoantigen+in+halothane+hepatitis.+Molecular+mimicry+of+trifluoroacetyl-lysine+by+lipoic+acid.">Identification of the dihydrolipoamide acetyltransferase subunit of the human pyruvate dehydrogenase complex as an autoantigen in halothane hepatitis. Molecular mimicry of trifluoroacetyl-lysine by lipoic acid.</a> European Journal of Biochemistry. 223 (3), 1035-1047 (1994).</li></ol>

该协议的优势在于其可重复性;因此，遵守建议的步骤至关重要。免疫原的配方对某些人来说可能是一个障碍;然而，我们已经使用文档中描述的表位重现了我们的模型，这消除了分离肝脏S100部分的需要。额外的表位或蛋白质很可能被改变，并在免疫接种后诱发肝炎;然而，我们描述了那些我们已经使用的蛋白质，并获得了可靠的结果。几种蛋白质已被证明在卤化麻醉剂暴露时是三氟乙酰化的。最有...

该方法的目的是描述在体内发展的药物诱导的自身免疫性肝炎的小鼠模型，并证明如何利用它来研究这种疾病的分子，免疫学和遗传基础。我们研究的长期目标是通过研究易感患者的DIH来揭示导致慢性肝脏炎症和损伤发展的机制。肝病和肝硬化是25至64岁成年人的第六大常见死因。特异质性 DILI，有时也称为药物诱发的自身免疫性肝炎 （DIH），是美国急性肝功能衰竭的第三大常见病因。DIH 是最常见的肝药物诱导的超敏化过程，见于约 9% 至 12% 的自身免疫性1 型肝炎患者。绝大多数 DIH 患者是女性 2，3，4。一种类型的DIH在给予卤化挥发性麻醉剂（例如异氟醚，七氟醚，去氟醚或氟烷）后在易感个体中发展。这些麻醉剂与其代谢的反应产物共价结合肝脏蛋白，从而产生能够引起过敏或自身免疫反应的新型自身抗原5。
对参与麻醉剂和任何形式的DIH发展的致病机制的研究以前一直受到缺乏密切模仿人类疾病诱导的动物模型的阻碍。我们开发了一种实验性的DIH小鼠模型，其特征类似于患者免疫介导的DILI。肝炎是通过用两种自身抗原之一的免疫诱导的，这两种自身抗原已被三氟乙酰氯（TFA）代谢物共价修饰，该代谢物是在细胞色素P450 2E1（CYP2E1）5的麻醉剂氧化代谢后形成的。一种自身抗原是肝细胞质 S100 肝脏部分，它是几种蛋白质6 的混合物，第二种自身抗原是 CYP2E1 的表位，由麻醉免疫介导的 DILI7 患者的血清识别。通过使用对实验性自身免疫性肝炎相对耐药的BALB / c小鼠，我们将我们的模型与C57Bl / 6J小鼠8中S100诱导的自身免疫性肝炎免疫模型区分开来。
由于其临床表现的多样性，DIH很难在患者中研究。转化实验模型提供了在体内和体外评估疾病发病机制的能力。目前，没有其他诱导DIH的替代方法可以在不使用动物的情况下充分检查体内或体外适应性或先天性免疫反应。此外，由于S-100或CYP2E1表位的三氟乙酰化似乎不会产生刺激性免疫原，并且我们通过用TFA改变的蛋白质免疫来诱导DIH，因此这些动物在免疫或其他程序之前不会接受乙醚，任何卤化麻醉剂，巴比妥酸盐或酒精，考虑到这些药物可能会改变我们正在研究的参数。即便如此，我们通过利用计算机模拟来确认我们发现的CYP2E1表位9 的结合偏好，并通过证明雌性BALB / c小鼠发展出更严重的DIH10来镜像涉及女性性别的人类DICH，从而减少了小鼠的使用量。
尽管 DIH 在患者中有不同的表现形式，并且在临床疾病研究中面临挑战，但反应性药物代谢物对天然蛋白的翻译后修饰是卤化麻醉剂11 之后的发病机制 DIH 中公认的关键机制。研究人员还承认CYP2E1是这一过程中的主要自原12，13。识别翻译后修饰的CYP2E1和其他肝脏蛋白的白细胞介素（IL）-4-上调CD4 + T细胞的作用是通过吸引嗜中性粒细胞，嗜酸性粒细胞和肥大细胞进入肝脏14来接受麻醉DIH的引发剂，并且该机制已在许多形式的DIH15，16中得到证实。诱导表达FoxP3的CD4 + CD25 + T细胞（Tregs）可降低DIH的严重程度，并且脾脏中这些细胞的相对缺乏会恶化DIH 10，7。因此，通过利用体内小鼠模型评估DIH在体内和体外的遗传，代谢和免疫机制，已经使理解DIH的大多数进展成为可能。
由于我们和其他研究人员已经发现了IL-4，嗜中性粒细胞和嗜酸性粒细胞在使用不同的小鼠模型启动DIH中的作用，我们相信这一观察结果支持了我们的论点，即无论使用DIH模型，肝炎和损伤都是由IL-4诱导的。我们方案的优势在于利用体内方法，包括雄性和雌性小鼠，以及重复组织学，CD4 + T细胞增殖测定和细胞因子。我们使用体外研究的优势在于，它们减少了所需的小鼠数量，同时提供了分离驱动DIH的细胞相互作用的方法。我们建议使用雄性和雌性小鼠，因为这降低了在解释结果时出现无意识偏倚的可能性，并增强了我们研究的转化潜力，因为DIH的发病率，患病率和严重程度在女性17中更高。我们建议从单一供应商处获得鼠标;然而，如果这是不可能的，从与转基因小鼠相同的供应商处获得窝交配对照或野生型小鼠。

<table border="0" cellpadding="0" cellspacing="0" style="width:572px;" width="573">
	<colgroup>
		<col />
		<col />
		<col />
		<col />
	</colgroup>
	<tbody>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">0.1% 2,4,6-trinitrobenzene sulfonic acid (TNBS)</td>
			<td style="width:71px;">ThermoFisher</td>
			<td align="right" style="width:119px;">28997</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">AKP Substrate Kit</td>
			<td style="width:71px;">BioRad</td>
			<td style="width:119px;">172-1063</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">BALB/c mice</td>
			<td style="width:71px;">Jackson</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">CellTrace&#8482; CFSE Cell Proliferation Kit</td>
			<td style="width:71px;">ThermoFisher</td>
			<td style="width:119px;">C34554</td>
			<td></td>
		</tr>
		<tr height="84">
			<td height="84" style="height:84px;width:216px;">CFA H37Ra</td>
			<td style="width:71px;">Becton Dickinson (Difco Bacto)</td>
			<td align="right" style="width:119px;">231131</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">FcR Blocking reagent</td>
			<td style="width:71px;">Milteyi</td>
			<td>130-092-575</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">General supplement</td>
			<td style="width:71px;">ThermoFisher</td>
			<td style="width:119px;">HPRG770</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">HepaRG&#8482; cells cryopreserved</td>
			<td style="width:71px;">ThermoFisher</td>
			<td style="width:119px;">HPR GC10</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Live/Dead Fixable Aqua Dead Cell stain kit&#160;</td>
			<td style="width:71px;">ThermoFisher</td>
			<td style="width:119px;">L34965</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">NaHC03</td>
			<td style="width:71px;">Millipore Sigma</td>
			<td style="width:119px;">S5761</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Percoll&#174;</td>
			<td style="width:71px;">Millipore Sigma</td>
			<td style="width:119px;">P1644-1L</td>
			<td></td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:216px;">Pertussis Toxin</td>
			<td style="width:71px;">List Biologicals</td>
			<td align="right" style="width:119px;">180</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Phosphate Buffered Saline pH 7.4</td>
			<td style="width:71px;">Various</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Pierce&#8482; Protease Inhibitor Mini Tablets, EDTA Free</td>
			<td style="width:71px;">ThermoFisher</td>
			<td align="right" style="width:119px;">88666</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Potassium Hydroxide</td>
			<td style="width:71px;">JT Baker</td>
			<td style="width:119px;">3140-01</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">S-ethyltrifluorothioacetate (S-ETFA)</td>
			<td style="width:71px;">Millipore Sigma</td>
			<td align="right" style="width:119px;">177474</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Slide-a-lyzer dialysis cassettes (10 K, 12 ml)</td>
			<td style="width:71px;">ThermoFisher</td>
			<td align="right" style="width:119px;">66810</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">UltraPure&#8482; SDS Solution, 10%</td>
			<td style="width:71px;">ThermoFisher</td>
			<td align="right" style="width:119px;">24730020</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Williams Media E, no phenol red</td>
			<td style="width:71px;">ThermoFisher</td>
			<td style="width:119px;">A1217601</td>
			<td></td>
		</tr>
	</tbody>
</table>

induction of drug-induced, autoimmune hepatitis in balb/c mice for the study of its pathogenic mechanisms

药物诱发的自身免疫性肝炎 （DIH） 是最常见的肝药物诱导的超敏化过程，见于约 9% 至 12% 的自身免疫性肝炎患者。绝大多数 DIH 患者是女性。由于缺乏模仿人类疾病的动物模型，这些性别差异的患病率的潜在机制尚不清楚。即便如此，人们普遍认为潜在的机制与人类白细胞抗原单倍型和性激素有关。相比之下，使用DIH小鼠模型，我们发现IL-4启动的CD4 + T细胞直接针对细胞色素P450 2E1的表位诱导嗜中性粒细胞，巨噬细胞和肥大细胞流入雌性BALB / c小鼠的肝脏。使用该模型，我们还表明IL-33诱导的FoxP3 +调节性T细胞在雌性和雄性小鼠中具有针对DIH的保护作用。该DIH模型是通过用CYP2E1表位对小鼠进行免疫诱导的，该表位已与与DIH相关的药物代谢物共价改变。这种表位被 DIH 患者识别。我们的方法诱导稳健且可重复的肝炎和自身抗体，可用于研究DIH的发病机制。虽然体内研究在处理不当时会在小鼠中引起过度的疼痛和痛苦，但体内模型的优点是能够评估大量小鼠的疾病发病机制。此外，可以使用侵入性程序研究改变的肝脏蛋白质的生物效应。在实验设计中添加体外研究允许在细胞水平上进行快速重复和机械分析。因此，我们将演示我们的模型方案以及如何利用它来研究DIH的体内和体外机制。

图1所示用于诱导DIH的免疫计划代表了颈部底部（第0天）和尾巴底部（第7天）所需的两次免疫接种。图2显示了使用CFSE响应CYP2E1，JHDN5，CYP2E1表位和麻醉剂的三氟乙酰基（TFA）代谢物在第14天获得的代表性增殖数据。图3显示了第14天获得的诱导CD4 + CD25 + FoxP3 + Tregs的门控策略和代表性流式细胞术分析。图4显示?...

Watch this Scientific Journal Video about Induction of Drug-Induced, Autoimmune Hepatitis in BALB/c Mice for the Study of Its Pathogenic Mechanisms at JoVE.com

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

我们描述了BALB / c小鼠中的体内免疫，转化性肝炎模型，可用于研究药物诱导的自身免疫性肝炎的发病机制，包括该疾病中看到的性别差异。我们将描述该模型如何使用体内和体外实验技术演示可重复的分析。

诱导BALB/c小鼠药物诱导的自身免疫性肝炎研究其致病机制

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

Research

JoVE Journal

Immunology and Infection

Preparation of Mouse Pituitary Immunogen for the Induction of Experimental Autoimmune Hypophysitis

鼠标垂体免疫原的制备诱导实验性自身免疫性垂体炎

Autoimmune hypophysitis is a chronic inflammation of the pituitary gland caused or accompanied by autoimmunity1.  It has traditionally been considered a ...

科研

免疫与感染

Induction of Experimental Autoimmune Hypophysitis in SJL Mice

SJL小鼠诱导实验性自身免疫性垂体炎

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

Determination of Tolerable Fatty Acids and Cholera Toxin Concentrations Using Human Intestinal Epithelial Cells and BALB/c Mouse Macrophages

使用人类肠道上皮细胞和BALB / C小鼠巨噬细胞的容忍脂肪酸和霍乱毒素浓度的测定

The positive role of fatty acids in the prevention and alleviation of non-human and  human diseases have been and continue to be extensively documented. ...

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

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

髓鞘少突胶质细胞糖蛋白（MOG<sub&gt; 35-55</sub&gt;）诱导实验性自身免疫性脑脊髓炎（EAE）的C57BL / 6小鼠

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

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

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

诱导小鼠肠道炎症由效应CD4的过继转移<sup&gt; +</sup&gt; CD45RB<sup&gt;高</sup&gt; T细胞免疫缺陷小鼠

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

Immunofluorescence to Monitor the Cellular Uptake of Human Lactoferrin and its Associated Antiviral Activity Against the Hepatitis C Virus

免疫监视人乳铁蛋白的细胞摄取及其相关抗病毒活性抗丙型肝炎病毒

Immunofluorescence is a laboratory technique commonly used to study many aspects of biology. It is typically used to visualize the distribution and/or ...

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

在免疫细胞在不同组织与疾病相关的分布和小鼠实验评价性自身免疫性脑脊髓炎的诱导

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

Development of a Hepatitis B Virus Reporter System to Monitor the Early Stages of the Replication Cycle

乙型肝炎病毒记者系统的开发监控复制周期的早期阶段

Currently, it is possible to construct recombinant forms of various viruses, such as human immunodeficiency virus 1 (HIV-1) and hepatitis C virus (HCV), ...

Increased Recovery Time and Decreased LPS Administration to Study the Vagus Nerve Stimulation Mechanisms in Limited Inflammatory Responses

增加了恢复时间，并减少LPS管理研究迷走神经刺激机制的有限炎症反应

Inflammation is a local response to infection and tissue damage mediated by activated macrophages, monocytes, and other immune cells that release ...

Induction of an Inflammatory Response in Primary Hepatocyte Cultures from Mice

从小鼠在原代肝细胞培养物的炎性反应的诱导

The liver plays a decisive role in the regulation of systemic inflammation. In chronic kidney disease in particular, the liver reacts in response to the ...