Name: chronic, acute, and reactivated hiv infection in humanized immunodeficient mouse models
Uploaded: 2019-12-03T12:30:00
Description: Watch this Scientific Journal Video about Chronic, Acute, and Reactivated HIV Infection in Humanized Immunodeficient Mouse Models at JoVE.com

这项工作得到了IHV临床部门内部资金对JCZ的支持。

在这项工作中，所有动物护理和程序都按照马里兰大学医学院机构动物护理和使用委员会（IACUC）审查和批准的协议进行（协议号1018017、1018018和0318009）。
1. 人类CD34=新生小鼠的HSC移植
<ol><li>始终使用一次性个人防护设备 （PPE），包括无菌磨砂、手套、专用鞋、鞋套、面罩、护目镜、头发/胡须发动机罩和无菌实验室外套。</li>
	<li>在 10 mL 的 RPMI 1640 介质 10% FBS 中，?...

<ol>
	<li>Shultz, L. D., Ishikawa, F., Greiner, D. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Humanized+mice+in+translational+biomedical+research.">Humanized mice in translational biomedical research.</a> Nature Reviews Immunology. 7, 118-130 (2007).</li><li>Koboziev, I., 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=Use+of+humanized+mice+to+study+the+pathogenesis+of+autoimmune+and+inflammatory+diseases.">Use of humanized mice to study the pathogenesis of autoimmune and inflammatory diseases.</a> Inflammatory Bowel Diseases. 21 (7), 1652-1673 (2015).</li><li>Ito, 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=NOD/SCID/&#947;cnull+mouse:+An+excellent+recipient+mouse+model+for+engraftment+of+human+cells.">NOD/SCID/&#947;cnull mouse: An excellent recipient mouse model for engraftment of human cells.</a> Blood. 100 (9), 3175-3182 (2002).</li><li>Ishikawa, F., 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=Development+of+functional+human+blood+and+immune+systems+in+NOD/SCID/IL2+receptor+{gamma}+chain(null)+mice.">Development of functional human blood and immune systems in NOD/SCID/IL2 receptor {gamma} chain(null) mice.</a> Blood. 106 (5), 1565-1573 (2005).</li><li>Kim, K. 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=A+Simple+Mouse+Model+for+the+Study+of+Human+Immunodeficiency+Virus.">A Simple Mouse Model for the Study of Human Immunodeficiency Virus.</a> AIDS research and human retroviruses. 32 (2), 194-202 (2016).</li><li>Wunderlich, 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=AML+xenograft+efficiency+is+significantly+improved+in+NOD/SCID-IL2RG+mice+constitutively+expressing+human+SCF,+GM-CSF+and+IL-3.">AML xenograft efficiency is significantly improved in NOD/SCID-IL2RG mice constitutively expressing human SCF, GM-CSF and IL-3.</a> Leukemia. 24 (10), 1785-1788 (2010).</li><li>Billerbeck, E., 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=Development+of+human+CD4+FoxP3++regulatory+T+cells+in+human+stem+cell+factor-,+granulocyte-macrophage+colony-stimulating+factor-,+and+interleukin-3-expressing+NOD-SCID+IL2R&#947;null+humanized+mice.">Development of human CD4+FoxP3+ regulatory T cells in human stem cell factor-, granulocyte-macrophage colony-stimulating factor-, and interleukin-3-expressing NOD-SCID IL2R&#947;null humanized mice.</a> Blood. 117 (11), 3076-3086 (2011).</li><li>Coughlan, A. 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在开发用于人性化的免疫缺陷小鼠菌株方面取得了重要进展，根据研究兴趣1，可以使用许多不同的选择。这里提供了一个一般方案，用于NS +链空小鼠和基因相似的菌株的人性化，用于三种不同的模型，用于研究艾滋病毒感染。在第一种实验方法中，辐照的新生小鼠被注射人类CD34+HSC，可以从脐带血、胎儿肝脏或动员的外周血3，21。<sup class="xref"...

人性化的NOD/SCID/白细胞素（IL）-2受体+链空（以下简称huNS+链空）小鼠模型已被广泛用于研究感染、自身免疫和癌症的发病机制，以及药物和基于人体细胞的疗法1、2的临床前研究。这些小鼠基于非肥胖糖尿病（NOD）背景，在IL-2受体β链位点（IL-2、IL-4、IL-7、IL-9、IL-15和IL-21）的共发β链上具有杀伤性突变和靶向突变，这在小鼠T-、B-和自然杀伤细胞（NK）的发育中引起严重损害。因此，它们支持人体组织、人类CD34+造血干细胞（HSCs）和人类外周血单核细胞（PBMC）3、4、5的移植。 此外，人类造血因子的转基因表达，如干细胞因子（SCF）、粒细胞因子/巨噬细胞菌群刺激因子（GM-CSF）和IL-3促进人类骨髓群6、7、8的移植。
在HIV研究中，描述了几种HuNS-链空小鼠模型，这些模型在小鼠菌株、使用的人类细胞类型、用于移植的组织类型以及细胞的来源（即健康与） 之间有所不同。HIV感染者）9日，10日。原始菌株，然而，被广泛使用，由于高水平的人体细胞移植和病毒复制后感染与参考HIV菌株11，12，13。具有人类造血因子（如NOG-EXL或NSG-SGM3）或植入人类肝脏和胸腺组织（骨髓-肝-胸腺[BLT]小鼠）的类似免疫缺陷小鼠菌株，对于评估骨髓群在抗HIV免疫反应中的作用、HIV对这些组织的影响及其作为病毒库的参与非常有用。此外，一些具有转基因表达的人类白细胞抗原（HLA）分子，以及BLT小鼠，可用于研究T细胞对HIV感染的反应16，17。
一般来说，在这些小鼠中，人性化取决于细胞来源、分娩途径（腹内、内肝、静脉内、心脏内）和小鼠年龄，在移植时为18、19、20。关于细胞来源，从脐带血、胎儿肝脏或动员的外周血中提取的人类CD34+HSC可注射在新生儿或幼鼠3、21。此外，成人β链空小鼠可以通过注射PBMC（这里称为hu-PBL-NS+链空小鼠）进行人体化，允许这些细胞在血液、继发性淋巴器官和发炎组织22、23、24中进行时间循环。
本文介绍了建立HUNS+链空鼠模型以研究HIV感染的详细方案。第一种是慢性模型，其中从健康供体脐带血中提取的人类CD34+HC被注射到新生小鼠中，随后在人类免疫系统重组14周后感染参考HIV菌株。此模型允许在感染后长达 36 周对小鼠进行监测。第二个模型是急性模型，其中从健康捐赠者中提取的PBMC注射到成年NS+链空小鼠中，随后在小鼠中人类T细胞扩张3周后感染参考HIV菌株。最后，第三个模型是重新激活模型，其中从受艾滋病毒感染的捐赠者在抑制性抗逆转录病毒疗法（ART）下获得的PBMC注射到成人NS+链空小鼠中。在这种情况下，无药物环境允许病毒重新激活和增加病毒载量。后两种模型允许在移植后长达 9 周的监测。
总体而言，这三种模型对于病毒学研究、新药临床前研究以及HIV感染对全球免疫反应的影响评估都很有用。还必须考虑，使用感染艾滋病毒的人性化小鼠需要机构生物安全委员会（IBC）以及机构动物护理和使用委员会（IACUC）在进行任何实验之前进行审查和批准。这确保了该研究遵循所有内部和外部机构条例，规定使用危险生物材料和人道处理实验动物。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>0. 5 ml Microcentrifuge tubes</td>
			<td>Neptune</td>
			<td>3735.S.X</td>
			<td></td>
		</tr>
		<tr>
			<td>1. 5 ml Microcentrifuge tubes</td>
			<td>Neptune</td>
			<td>3745.S.X</td>
			<td></td>
		</tr>
		<tr>
			<td>10 ml Serologial pipetes</td>
			<td>stellar sceintific</td>
			<td>VL-4090-0010</td>
			<td></td>
		</tr>
		<tr>
			<td>15 ml conical tubes</td>
			<td>Stellar scientific</td>
			<td>T15-600</td>
			<td></td>
		</tr>
		<tr>
			<td>25 ml Serologial pipetes</td>
			<td>stellar sceintific</td>
			<td>VL-4090-0025</td>
			<td></td>
		</tr>
		<tr>
			<td>5 ml Serologial pipetes</td>
			<td>stellar sceintific</td>
			<td>VL-4090-0005</td>
			<td></td>
		</tr>
		<tr>
			<td>50 ml conical tubes</td>
			<td>Stellar scientific</td>
			<td>T50-600</td>
			<td></td>
		</tr>
		<tr>
			<td>ACK lysis buffer</td>
			<td>Quality biological</td>
			<td>118-156-101</td>
			<td></td>
		</tr>
		<tr>
			<td>Alcohol prep pads</td>
			<td>Fisher scientific</td>
			<td>06-669-62</td>
			<td>Sterile</td>
		</tr>
		<tr>
			<td>Anti-Human CD3 clone UCHT1</td>
			<td>Biolegend</td>
			<td>300439</td>
			<td>APC conjugated</td>
		</tr>
		<tr>
			<td>Anti-Human CD4 clone OKT4</td>
			<td>Biolegend</td>
			<td>317420</td>
			<td>AF488 conjugated</td>
		</tr>
		<tr>
			<td>Anti-Human CD45 clone 2D1</td>
			<td>Biolegend</td>
			<td>368522</td>
			<td>BV421 conjugated</td>
		</tr>
		<tr>
			<td>Anti-Human CD8 clone SK1</td>
			<td>Biolegend</td>
			<td>344710</td>
			<td>PerCP-Cy5.5 conjugated</td>
		</tr>
		<tr>
			<td>Biosafaty cabinet level 2</td>
			<td></td>
			<td></td>
			<td>If posible connected to an exauste chimeny when handling Isoflurane</td>
		</tr>
		<tr>
			<td>Bonnet</td>
			<td>Fisher scientific</td>
			<td>17-100-900</td>
			<td>Single use cap for basic protection</td>
		</tr>
		<tr>
			<td>Cavicide</td>
			<td>Metrex</td>
			<td>13-1000</td>
			<td>Surface desinfectant</td>
		</tr>
		<tr>
			<td>CD34+ cells</td>
			<td>Lonza</td>
			<td>2C-101</td>
			<td>As many vials available from a single donor</td>
		</tr>
		<tr>
			<td>Centrifuge</td>
			<td>Beckman</td>
			<td>65-6KR</td>
			<td></td>
		</tr>
		<tr>
			<td>Clear jar</td>
			<td>Amazon</td>
			<td>77977</td>
			<td></td>
		</tr>
		<tr>
			<td>Cotton gauze pad</td>
			<td>Fisher scientific</td>
			<td>22-415-468</td>
			<td>Sterile</td>
		</tr>
		<tr>
			<td>Disposable lab coats</td>
			<td>Fisher scientific</td>
			<td>19-472-422</td>
			<td></td>
		</tr>
		<tr>
			<td>EDTA micro tubes</td>
			<td>Greiner bio-one</td>
			<td>450480</td>
			<td></td>
		</tr>
		<tr>
			<td>Face Mask</td>
			<td>Fisher scientific</td>
			<td>17-100-897</td>
			<td></td>
		</tr>
		<tr>
			<td>FACS lysing solution</td>
			<td>BD</td>
			<td>340202</td>
			<td></td>
		</tr>
		<tr>
			<td>FBS premium HI</td>
			<td>Atlanta biologicals</td>
			<td>S1115OH</td>
			<td></td>
		</tr>
		<tr>
			<td>Ficoll</td>
			<td>GE health one</td>
			<td>17-1440-02</td>
			<td></td>
		</tr>
		<tr>
			<td>Flow cytometer</td>
			<td></td>
			<td></td>
			<td>We used FACS Aria II</td>
		</tr>
		<tr>
			<td>Flow cytometry tubes</td>
			<td>Falcon</td>
			<td>352054</td>
			<td>5 ml polystyrene and round bottom</td>
		</tr>
		<tr>
			<td>HIV BaL</td>
			<td></td>
			<td></td>
			<td>Prepared in our uQUANT core facility</td>
		</tr>
		<tr>
			<td>Human PBMCs</td>
			<td></td>
			<td></td>
			<td>HIV positive and negative volunteers</td>
		</tr>
		<tr>
			<td>Infrared warming pad</td>
			<td>Venet scientific</td>
			<td>DCT-25</td>
			<td>Temporary therapeutic warming pad for small animals</td>
		</tr>
		<tr>
			<td>Isentress (Raltegravir)</td>
			<td>Merck</td>
			<td>NSC 0006-0227061</td>
			<td>Antiretroviral medication to treat human immunodeficiency virus (HIV)-Integrase inhibitor</td>
		</tr>
		<tr>
			<td>Isoflurane</td>
			<td>Henry Schein</td>
			<td>NDC 11695-6776-2</td>
			<td></td>
		</tr>
		<tr>
			<td>Mark I irradiator</td>
			<td></td>
			<td></td>
			<td>Equipment belonging to university of Maryland</td>
		</tr>
		<tr>
			<td>Micro pipettes</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Microcentrifuge</td>
			<td>Eppendorf</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Mouse ear tags</td>
			<td>National Band &#38; Tag company</td>
			<td>1005-1L1</td>
			<td></td>
		</tr>
		<tr>
			<td>Natelson blood collection tubes</td>
			<td>Fisher scientific</td>
			<td>02-668-10</td>
			<td></td>
		</tr>
		<tr>
			<td>NOG-EXL</td>
			<td>Taconic</td>
			<td>HSCFTL-13395-F</td>
			<td></td>
		</tr>
		<tr>
			<td>NSG mice</td>
			<td>Jackson</td>
			<td>5557</td>
			<td>Time pregnant females for CD34 engraftment and Juveniles for PBMCs engraftment</td>
		</tr>
		<tr>
			<td>NSG-SGM3</td>
			<td>Jackson</td>
			<td>13062</td>
			<td></td>
		</tr>
		<tr>
			<td>Paraformaldehyde 16%</td>
			<td>Electron microscopy sciences</td>
			<td>15710</td>
			<td></td>
		</tr>
		<tr>
			<td>PBS 1X pH 7.4</td>
			<td>Gibco</td>
			<td>100-10-023</td>
			<td></td>
		</tr>
		<tr>
			<td>Petri dishes</td>
			<td>Fisher scientific</td>
			<td>08-757-28</td>
			<td></td>
		</tr>
		<tr>
			<td>Quantistudio qPCR machine</td>
			<td>Thermo</td>
			<td>QS3</td>
			<td></td>
		</tr>
		<tr>
			<td>Reagent reservoirs</td>
			<td>Costar</td>
			<td>4870</td>
			<td></td>
		</tr>
		<tr>
			<td>RPMI media 1640 1X</td>
			<td>Gibco</td>
			<td>11875-093</td>
			<td></td>
		</tr>
		<tr>
			<td>Shoe covers</td>
			<td>Fisher scientific</td>
			<td>17-100-911</td>
			<td></td>
		</tr>
		<tr>
			<td>Sterile disposable Gloves</td>
			<td>Microflex</td>
			<td>SUF-524</td>
			<td></td>
		</tr>
		<tr>
			<td>SuperScript II First-Strand Synthesis SuperMix</td>
			<td>Invitrogen</td>
			<td>10080-400</td>
			<td>cDNA synthesis</td>
		</tr>
		<tr>
			<td>Syringes 28-G x 1/2</td>
			<td>BD</td>
			<td>329-461</td>
			<td></td>
		</tr>
		<tr>
			<td>Syringes 29-G x 1/2</td>
			<td>BD</td>
			<td>324-702</td>
			<td></td>
		</tr>
		<tr>
			<td>Truvada (Emtricitabine and Tenofovir</td>
			<td>Gilead</td>
			<td>NDC 61958-0701-1</td>
			<td>Antiretroviral medication to treat human immunodeficiency virus (HIV)-Nicleoside analog-transcriptase inhibitor</td>
		</tr>
		<tr>
			<td>Trypan blue</td>
			<td>Sigma</td>
			<td>T8154</td>
			<td>Cell count and viability</td>
		</tr>
		<tr>
			<td>Vick Vaporub</td>
			<td>School health</td>
			<td>43214</td>
			<td>Ointment based on menthol and eucalyptus</td>
		</tr>
		<tr>
			<td>Water molecular biology grade</td>
			<td>Quality biological</td>
			<td>351-029-131</td>
			<td></td>
		</tr>
	</tbody>
</table>

chronic, acute, and reactivated hiv infection in humanized immunodeficient mouse models

人性化NOD/SCID/IL-2受体+链空小鼠重述人体免疫的一些特征，在传染病的基础和临床前研究中可以加以利用。本文介绍了三种用于研究HIV感染动态的人性化免疫缺陷小鼠模型。第一种是基于CD34的肝内注射-在新生小鼠的造血干细胞，这允许重组几个血液和淋巴组织封闭细胞，其次是感染与参考HIV菌株。此模型允许在感染后长达 36 周的监测，因此称为慢性模型。第二和第三个模型称为急性和再激活模型，其中外周血单核细胞在成年小鼠中腹中注射。在急性模型中，来自健康供体的细胞通过腹管内途径移植，然后感染参考HIV菌株。最后，在复活模型中，来自抗逆转录病毒疗法的受艾滋病毒感染捐赠者的细胞通过腹内途径进行移植。在这种情况下，鼠标中的无药物环境允许病毒重新激活和增加病毒载量。此处提供的方案描述了对HIV感染的人性化免疫缺陷小鼠模型的传统实验方法。

如上所述，在HSC注射后14周（慢性模型）或PBMC注射后3周（急性和再活化模型）时，小鼠通过流动细胞测定筛选人体细胞移植水平。图1A显示了用于评估1人CD45+细胞重组和2）CD4+和CD8+T细胞百分比的代表性门控策略。通常，移植水平（人类CD45+细胞的百分比）在CD34+HSC注射后为10%-80%，取决于注射和小鼠菌株的路线，除上述其他因素外...

Watch this Scientific Journal Video about Chronic, Acute, and Reactivated HIV Infection in Humanized Immunodeficient Mouse Models at JoVE.com

Chronic, Acute, and Reactivated HIV Infection in Humanized Immunodeficient Mouse Models

本文介绍了三种研究人类小鼠HIV感染动态的实验方法。前者允许研究慢性感染事件，而后者允许研究原发感染或病毒重新激活后的急性事件。

人类免疫缺陷小鼠模型中的慢性、急性和再激活HIV感染

Humanized NOD/SCID/IL-2 receptor &#947;-chainnull mice recapitulate some features of human immunity, which can be exploited in basic and pre-clinical ...

该程序描述了三种用于研究HIV感染动态的人类免疫缺陷小鼠模型。NSG人性化小鼠综述了人类免疫力的特征。在这种情况下，允许在临床前环境中研究慢性、急性和重新激活的艾滋病毒感染。展示细胞制备程序的将是桑德拉·梅迪纳-莫雷诺，我们集团的实验室研究主管。执行此程序时，请始终使用一次性个人防护设备，包括无菌磨砂、手套、专用鞋、鞋套、面罩、护目镜、头发和胡须盖以及无菌实验室外套。对于慢性模型，在经过认证的生物安全柜下，在 10 毫升 RPMI 1640 介质中重新发送 100 万个冷冻 CD34® 人类干细胞，并保留无菌条件。使用悬浮液的10微升来计数和检查细胞的生存能力，通过 Trypan Blue 排除染色在血细胞计。然后在室温下以400倍g离心细胞悬浮液15分钟。丢弃上清液，在所需的浓度下在冷1X PBS中重新暂停细胞。将细胞放在冰上直到注射。双手之间擦干净的床上用品，以掩盖接受者幼崽身上任何其他异味。然后将幼崽放入无菌的 100 平方毫米培养皿中，以及来自饲养员笼中的少量床上用品。将 Petri 盘放入干净的运输笼中，然后将笼子放入 Carboy 运输笼中，以保护幼崽免受直接光线和噪音的干扰。在运输到辐照室时，用盖垫盖住运输笼，避免动物暴露在环境中。按照文本协议中描述的麻醉后，准备通过肝注射进行包。为了尽量减少鼠标约束的时间，让一名调查员用干细胞悬浮液加载注射器，让另一名调查员抱住幼崽并进行注射。将 CD34® 人体干细胞悬浮液的 50 微升装入注射器，在经过认证的生物安全柜下用连接针插入注射器。用拇指和食指约束幼崽，用70%的酒精清洁注射部位。使用浅针角将50微升的细胞直接送入肝脏，这样才能完全刺穿肝脏。在20摄氏度下为啮齿动物使用红外加热垫预热，以确保幼崽不会过度加热。将幼崽放在用无菌纱布覆盖的培养皿上一至五分钟，以便恢复。在将幼崽归还给父母之前，请立即使用拇指和食指涂抹少量薄荷醇和桉树软膏，以避免吃人或排斥幼崽。每天检查笼子，寻找任何移植与宿主疾病的迹象，在幼崽，如干燥的皮肤，没有喂养，皮疹，或脱发。在三周大时把幼崽打奶，把它们关在不同的笼子里。通过14周大的流动细胞切除术验证周围血液的内皮。显示了一种代表性的浇注策略，用于评估人类CD45+细胞重建和CD4+和CD8+T细胞的百分比。移植的成功率在80%至100%之间，使用来自急性模型的健康供体或接受抗逆转录病毒治疗的HIV感染患者为再活化模型产生的人类外周血单核细胞。将全血第15毫升，以5毫升的无菌密度梯度介质放入50毫升锥形管中。在室温下以 400 倍 g 离心 30 分钟，不休息，避免布衣与密度梯度介质混合。仔细收集密度梯度介质和上细之间单核细胞的分数。将布衣转移到含有 10 毫升 1X PBS 的 15 毫升离心管中。在室温下以300倍g离心10分钟。丢弃上流剂，通过与添加到颗粒细胞中的五毫升CK解菌缓冲液进行解化，去除剩余的红血球。在室温下孵育四分钟。再次离心后，丢弃上经剂，再用 10 毫升 1X PBS 或 RPMI 1640 介质重新暂停。使用细胞悬浮液的10微升来计算细胞，并检查通过血细胞计中的 Trypan Blue 排除染色的可行性。通常，每毫升血液获得100万到200万个细胞，其生存能力超过95%。离心后，像以前一样，丢弃上一提液，并调整细胞数到所需的浓度。将 200 个 1X PBS 微升中的 350 万个外周血单核细胞装入经认证的生物安全柜下带连接针的注射器中。将鼠标从笼子上取下，然后用尾巴握住它，这样它就可以抓住网眼，向后轻轻牵引。然后将食指和拇指放在动物的肩膀上，抓住颈部松动的皮肤，用中指稳定背部。向后滑动鼠标头，使鼠标背面位于其头部上方。这使得腹腔中的内脏向后置换，并降低注射期间刺穿内脏器官的风险。用 70% 酒精清洁注射部位。通过腹壁穿透准备好的注射器，在注射细胞前吸气。如果任何材料被吸入，请取出注射器并丢弃。否则，在取出注射器并丢弃注射器之前，将细胞缓慢注射到内腔中。将动物返回到其笼子，并在注射后三周内通过流动细胞测量验证周围血液的含量。通过耳朵标记识别小鼠。仔细观察这些实验中使用的小鼠，每次手术后每天两次出现危难症状。然后按照文本协议中所述进行艾滋病毒注射。在艾滋病毒注射之后，在重新激活模型中具有类似动力学的慢性和急性模型中，在感染后两到三周通常可检测到血浆病毒载量迅速增加。病毒载量的增加与CD4：CD8比率的下降相吻合。这些变化在对照小鼠中未观察到。值得注意的是，在人性化PBMC NSG成人小鼠模型中，观察到CD4：CD8比率的初始反转，并沿监测时间进行重组。当对艾滋病毒感染小鼠进行抗逆转录病毒治疗时，CD4：CD8比率抑制病毒载量和恢复情况如预期的那样达到与未受感染对科小鼠类似的水平。通常，经过两到三周的治疗后，在慢性、急性和重新激活模型中观察到病毒载量减少和CD4：CD8比率增加。使用推荐的辐照剂量，避免致命过量或排斥移植细胞。在注射过程中不要完全刺穿肝脏，并使用床上用品来掩盖大坝会认为异味的气味。这三种模型允许测试抗病毒化合物、抗HIV抗体或其他影响病毒复制的生物物质。此外，它们还允许艾滋病毒免疫治疗的适应性转移实验。NSG人性化小鼠可以部分或全部与人体细胞重组，以对病原体或药物作出反应，模仿人类免疫系统的复杂性。所有基本的生物措施都需要严格遵守，因为NSG人性化小鼠可能同时感染穆林和人类病原体，特别是在使用HIV病毒操纵的情况下。

Research

JoVE Journal

Immunology and Infection

Genotypic Inference of HIV-1 Tropism Using Population-based Sequencing of V3

V3的HIV - 1取向的基因型推理使用人口为基础的测序

Background: Prior to receiving a drug from CCR5-antagonist class in HIV therapy, a patient must undergo an HIV tropism test to confirm that his or her ...

科研

免疫与感染

In vivo Imaging Method to Distinguish Acute and Chronic Inflammation

In vivo Imaging Method to Distinguish Acute and Chronic Inflammation

在体内成像方法区分急性和慢性炎症

Inflammation is a fundamental aspect of many human diseases. In this video report, we demonstrate non-invasive bioluminescence imaging techniques that ...

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 ...

Humanized Mouse Model to Study Bacterial Infections Targeting the Microvasculature

人源化的小鼠模型来研究细菌感染瞄准微血管

Neisseria meningitidis causes a severe, frequently fatal sepsis when it enters the human blood stream. Infection leads to extensive damage of the blood ...

Rapid Screening of HIV Reverse Transcriptase and Integrase Inhibitors

HIV逆转录酶和整合酶抑制剂的快速筛选

Although a number of anti HIV drugs have been approved, there are still problems with toxicity and drug resistance. This demonstrates a need to identify ...

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 ...

Mouse Models Of Helicobacter Infection And Gastric Pathologies

Mouse Models Of Helicobacter Infection And Gastric Pathologies

幽门螺杆菌感染与胃病理的小鼠模型

Helicobacter pylori is a gastric pathogen that is present in half of the global population and is a significant cause of morbidity and mortality in ...

Humanized NOD/SCID/IL2r&#947;null (hu-NSG) Mouse Model for HIV Replication and Latency Studies

Humanized NOD/SCID/IL2rÃƒÆ’Ã…Â½Ãƒâ€šÃ‚Â³null hu-NSG Mouse Model for HIV Replication and Latency Studies

人性化的 nod/scid/il2γ空 (hu-nsg) 小鼠 hiv 复制和延迟研究模型

Ethical regulations and technical challenges for research in human pathology, immunology, and therapeutic development have placed small animal models in ...

CRISPR-Cas9-based Genome Engineering to Generate Jurkat Reporter Models for HIV-1 Infection with Selected Proviral Integration Sites

基于 crispr-cas9 的基因组工程, 用于生成 hiv-1 感染与选定的证明病毒整合点的 jurkat 报告模型

Human immunodeficiency virus (HIV) integrates its proviral DNA non-randomly into the host cell genome at recurrent sites and genomic hotspots. Here we ...

Establishment of the Dual Humanized TK-NOG Mouse Model for HIV-associated Liver Pathogenesis

建立与艾滋病毒相关的肝脏发病机制的双人性化TK-NOG小鼠模型

Despite the increased life expectancy of patients infected with human immunodeficiency virus-1 (HIV-1), liver disease has emerged as a common cause of ...