We would like to thank the vivarium manager and staff at San Diego State University for support with animal husbandry. During this work, K.A.P. was supported by an ARCS scholarship and a fellowship from the Inamori Foundation. K.S.D. is supported by an R01 grant, NS051247, from the National Institutes of Health.

所有的动物工作是经实验室动物护理在州立圣迭戈大学办公室和下接受兽医标准进行。雌性小鼠，年龄8 - 16周，用于该方法的发展。 1.β雌二醇的制备和腹腔注射<ol><li>测量出的称量纸而穿着适当的个人防护装备（PPE）β雌二醇（0.5毫克/鼠标）。 注意:β雌二醇可通过皮肤和粘膜表面吸收。 </li><li>转移β雌二醇到15毫升锥形管中，涡旋，直到所有的团块被去除和β雌二醇是细粉。拟定香油（100微升/小鼠）到10毫升注射器。注射器过滤芝麻油成用0.45微米的过滤器含有β雌二醇的15-ml锥形管中。涡15毫升锥形管，直到将β雌二醇处于芝麻油一个均匀的悬浮液。 </li><li>卷起β-EStradiol悬挂到一个新的10毫升注射器。随着18克，1项。针，等份加入100μl悬浮液加入到1毫升的结核菌素注射器。准备每只小鼠一个针筒。将一个新的，无菌26 G，½英寸的针上的每个结核菌素注射器。 </li><li>施用0.5毫克悬浮于100μl麻油（5毫克/毫升），以每只小鼠24细菌接种之前小时的β雌二醇。注入每只小鼠的腹腔内，在下腹象限，只需向右侧或左侧的中线，如前所述28。 注:注射部位的无清洁或削波是必要的。 </li></ol> 2.阴道接种GBS <ol><li>接种前一天，在37℃下生长的兴趣，如A909（血清型Ia）的GBS菌株的5毫升过夜液体培养，在Todd Hewitt肉汤（THB）。 </li><li>亚文化的GBS过夜培养按1:10的体积到新鲜的泰铢，并在37℃孵育。成长细菌对数中期（OD 600 = 0.4-0.5）。注意:这将通常需要2 - 3小时，视GBS的应变。 </li><li>传送传代培养到无菌的15毫升锥形管中，沉淀的细菌以3000×g下5分钟。吸出上清液。重悬在200μl的无菌磷酸盐缓冲盐水（PBS）的细菌沉淀。 </li><li>使用再悬浮颗粒，带来3 - 5毫升的PBS（1毫升，每10只）恰好OD 600 = 0.4在新的5毫升培养管。这将是〜1×10 8菌落形成单位（CFU）/ ml的浓度。在3000×g下转移到一个新的15毫升锥形管，并重新沉淀菌5分钟。吸出上清液。 </li><li>重悬在PBS中的沉淀在1/10初始体积。例如，如果在三毫升OD 600 = 0.4粒化，然后重悬它在300微升PBS中。注意:这是用于动物接种的最终菌悬液（约1×109 CFU /毫升）。 </li><li> - [Reserve 50微升此悬浮液连续稀释的和电镀THB琼脂以确定确切的接种物。 </li><li>接种每个小鼠用10微升的最终细菌悬浮液，使1×10 7 CFU给药于每只小鼠。 <ol><li>接种，通过在处理程序的拇指和食指，然后固定尾部之间的颈背保护的皮肤松弛手动抑制鼠标，如前面28所描述。 </li><li>卷起GBS的10微升在步骤2.6制备成200微升凝胶装载枪头。插入尖端5至10毫米到阴道腔和分配所述10微升的接种物。 注:凝胶放入提示优于标准的200微升提示，以最大限度地减少器官损伤或伤害的危险，尤其是在年轻的或更小的小鼠。 </li><li>紧接着接种，松开颈背和提升鼠标的后端，由尾以及w提起鼠标烯王在坚硬的表面约1分钟的前爪。 </li><li>目视检查接种任何回流阴道口。如果观察到回流，新鲜枪头可以用于操纵或放大阴道口，便于回流摄取进入管腔。此外，逆流可能经由吸移管吸出，并重新接种。 注意:如果施用外用剂，益生菌微生物，或感兴趣的蛋白质，一个体积多达20微升在生理缓冲液可在阴道给予。 </li></ol></li></ol> 3.抽汲阴道流明量化GBS负载<ol><li>通过加入100微升的PBS制备每鼠1.5毫升微量离心管中。之前，为了擦拭，预湿的PBS拭子。 </li><li>抑制鼠标按步骤2.7.1描述插入擦拭器10毫米，伸入阴道腔内。轻轻顺时针4倍旋转拭子和4倍逆时针，轻轻施加压力，以阴道壁。</li><li>转移棉签1.5 ml离心管用100微升的PBS。电镀前，涡旋〜15秒，从拭子释放细菌的离心管中。每个制造商的说明制备差培养基琼脂平板10000:连续稀释在PBS中的每个样品和板20微升稀释液1:10至1。孵育所述板在37℃下24小时。 GBS菌落将出现在颜色无论是明亮的粉红色或紫红色。其它内源性菌群将受到抑制，或将显示为蓝色，白色或灰色菌落。 </li></ol> 4.收集阴道灌洗液<ol><li>如步骤2.7.1所述抑制鼠标。用200微升凝胶装载枪头，吸管20微升的PBS到阴道腔内。轻轻吸取整个卷上下4倍的管腔内，然后撤回在同一枪头的整个体积。注:如果灌洗液是厚厚的粘液，一个标准的200微升枪头，可用于收集最终灌洗液。 </li><li>如果储蓄细胞因子分析和定量CFU灌洗液，分装到0.7毫升离心管中。如果确定发情的阶段，分配至少5微升灌洗液到载玻片并观察下放大100倍的单元上的光学显微镜。注:对于发情阶段的例子， 见图1。 </li></ol> 5.组织解剖和同质化<ol><li>对于每个小鼠，制备三个2毫升螺旋盖管（一个用于每个组织:阴道，子宫颈和子宫）。填充每个管具有足够（0.4 - 0.5克）1.0毫米氧化锆珠，以覆盖管的圆锥形底部。 </li><li>高压灭菌之前收集组织在121℃，30分钟，尤其是如果量化细菌负荷制备管中。加入500微升无菌PBS至每个管。高压灭菌和记录以供将来参考，计算回收组织重量称量后，每管。 </li><li&gt;牺牲使用经批准的机构的方法，例如CO 2窒息和颈脱位鼠标。喷向下用70％乙醇的腹侧的腹部。随着无菌剪刀，打开腹盆腔，提起背部皮肤和腹部肌肉，并移动从而使生殖道露出的肠子。 </li><li>消毒用70％乙醇的剪刀，必要擦拭干净，并切成两个子宫角子宫体和卵巢之间的中点的长度。用剪刀和镊子，独立的内脏脂肪，膜和膀胱从生殖道远，尾部移动。 </li><li>如步骤5.4中所述消毒剪刀。横向切割阴道尽可能靠近外阴尽可能生殖道从主体分离。提起并取出完整的生殖道，并将其放置在无菌培养皿中。 </li><li>使用新的刀片，在一个横向切割从宫颈分离子宫。消毒刀片用70％EtOH中，然后从一个横向切阴道分离子宫颈。注意:有将仍然附着在子宫颈的子宫组织的最小量。会有仍然附着在子宫颈阴道组织的最小量。 </li><li>使用无菌镊子，传送每个组织到含有PBS中并均质珠粒各自的2毫升试管中。清洁用70％乙醇钳子在处理各组织之间。 </li><li>称重各2毫升螺丝帽管中并减去管的原重量来确定组织重量。组织重量一般变化在20 - 100毫克。紧密地密封螺旋盖管和在组织匀浆器以最大速度均化1分钟的组织中。 </li><li>为了量化细菌量，连续稀释25微升组织匀浆和板块稀释1:10至1:在鉴别培养基琼脂平板10,000。如步骤3.3中所述孵育所述板。要存储细胞因子定量样品，冷冻组织匀浆 -20℃。 </li></ol>

作者什么都没有透露。

进一步的与主机的环境内的主机和其他微生物都GBS相互作用的认识的提高，需要的动物模型。这部作品描述了在小鼠建立阴道GBS定植的技术方面。该协议实现小鼠&gt; 90％定植不使用麻醉剂的接种细菌或收集拭子样品，免疫抑制剂，使定植，阴道预清洗，或添加剂增稠接种物。此外，该模型表明健壮再现性，具有适度的间实验变异GBS持久的长度和细菌负担。在这项研究中证明了代表性结果是独立的...

无乳链球菌 ，B组链球菌 （GBS），是一个封装的，革兰氏阳性细菌，其经常从肠道和健康成人的泌尿生殖道中分离。在20世纪70年代，GBS成为感染新生儿死亡率的主要代理，拥有超过7000例新生儿疾病每年1。早发性GBS病（EOD）发生在第一个小时内或生命天，出现肺炎或呼吸窘迫，常发展成败血症，而晚发性疾病（LOD）几个月后随之而来，并与菌血症，频繁地呈现进入脑膜炎2。截至2002年，该中心疾病控制和预防建议在妊娠后期和分娩期预防性使用抗生素（IAP），以GBS阳性母亲1 GBS定植阴道普遍筛查。尽管早发性疾病的减少到大约1000例，美国每年因IAP，GBS仍然是早发性新生儿败血症的主要原因，迟发发生不受影响1。无论是在宫内，分娩过程中，甚至迟发性的情况下，新生儿暴露于GBS需要生存，横向通过大量的主机环境和障碍，免疫逃避，并在脑膜炎，高度管制的血 - 的交接的情况下，脑屏障2。新生儿中的这些有毒相互作用的上游是产妇阴道的初始定植。产妇阴道GBS定植率从8-18％，在发达国家和发展中国家，12.7％，3,4估计的平均增长率。怀孕期间阴道内的定植GBS可能是恒定的，间歇性的，或短暂在个别妇女中5性质。有趣的是，产妇年龄&gt;36年与持续定植6有关。为GBS定植阴道大量生物和社会经济风险因素已经确定。生物因素包括胃肠道GBS殖民化和缺乏肠道内乳酸菌 。然而，种族，肥胖，卫生和性活动也与GBS阴道托架7相关联。 虽然臭名昭著，导致新生儿感染，GBS也导致了各种感染的孕产妇围产期都和产后的。 GBS托架在女性阴道炎8呈递增加，并且在某些情况下，甚至可能是疾病实体9。此外，孕期生殖道GBS的提升可能会导致羊膜腔内感染或绒毛膜羊膜炎10。此外，在高达怀孕3.5％，GBS传播到膀胱引起尿路感染或无症状菌11。怀孕期间GBS菌与产热，绒毛膜羊膜炎，早产，和prematur的风险增加有关膜12电子破裂。两者合计，GBS的阴道内的存在被链接到多个宿主组织的感染，并从这个利基消除GBS的能力是必要为孕产妇和新生儿的健康。 直到最近，大多数的工作检查与宫颈道GBS相互作用被限制为体外细胞模型13-15。 这些体外实验已表明，对于粘附重要细菌因素，包括表面蛋白例如一个菌毛和富含丝氨酸重复17,18，以及双组分调控系统15,19和阴道上皮的对全球转录反应GBS 19。然而，为了充分阐明阴道内的宿主 - 微生物相互作用，一个强大的动物模型是必要的。早期的工作表明，GBS可以从接种小鼠20,21和大鼠的阴道内被恢复&lt;SUP&gt;在两个怀孕和未怀孕的条件22。 2005年，短期GBS定植阴道小鼠为蓝本，研究噬菌体裂解酶的功效在24小时的时间内23治疗阴道GBS。事隔多年，长期阴道GBS定植小鼠模型的开发研究规范GBS持久主机和细菌因素。这种模式已经确定促进殖民GBS众多因素，包括地面附属物17,18和GBS双组分系统19,24。这种模式对宿主反应机制19,25的识别作出了贡献，是用来测试几种治疗策略，包括免疫调节肽26和益生菌27。该协议使接种GBS入小鼠阴道并随后跟踪殖民化和收集样品用于进一步分析的必要的指导。

<table><tbody><tr><td>Sesame oil&amp;nbsp;</td><td>Sigma Aldrich</td><td>S3547-250ML</td><td></td></tr><tr><td>&amp;beta;-Estradiol&amp;nbsp;</td><td>Sigma Aldrich</td><td>E8875-1G</td><td>CAUTION: Wear appropriate PPE. &amp;beta;-estradiol can be absorbed through the skin and mucosal surfaces.&amp;nbsp;</td></tr><tr><td>200 &amp;mu;l gel loading pipette tips&amp;nbsp;</td><td>USA Scientific</td><td>1252-0610</td><td></td></tr><tr><td>Urethro-genital, sterile, calcium alginate swabs</td><td>Puritan</td><td>25-801 A 50</td><td></td></tr><tr><td>CHROMagar StrepB</td><td>DRG International</td><td>SB282</td><td></td></tr><tr><td>Todd Hewitt Broth</td><td>Hardy Diagnostics</td><td>7161C</td><td></td></tr><tr><td>18 G, 1.5 inch needles</td><td>BD</td><td>305199</td><td></td></tr><tr><td>26 G, 0.5 inch needles</td><td>BD</td><td>305111</td><td></td></tr><tr><td>10 ml syringes</td><td>BD</td><td>309604</td><td></td></tr><tr><td>1 ml syringes</td><td>BD</td><td>309659</td><td></td></tr><tr><td>0.45 &amp;mu;m PVDF syringe filters</td><td>Whatman</td><td>6900-2504</td><td></td></tr><tr><td>Dulbecco&#039;s Phosphate-Buffered Salt Solution 1x</td><td>Corning</td><td>21-031-CV</td><td></td></tr></tbody></table>

a murine model of group b streptococcus vaginal colonization

无乳链球菌 （B组链球菌 ，GBS），是人体胃肠道和10个阴道的革兰氏阳性，无症状殖民者-成人的30％。在免疫受损的个体，包括新生儿，孕妇和老年人，GBS可以切换到病原体侵入引起败血症，关节炎，肺炎和脑膜炎。由于GBS是新生儿的领先细菌病原体，目前预防由妊娠后期筛查GBS阴道定植和GBS阳性母亲的后续围产期抗生素治疗。重GBS阴道负担为两个新生儿疾病和定植的危险因素。不幸的是，鲜为人知的是，主机和促进或允许GBS定植阴道细菌因素。这个协议描述了一种技术，使用单β雌二醇预处理和每日取样，以确定细菌洛阿建立持久的GBS阴道定植ð。它进一步详细信息的方法来管理其它疗法或感兴趣的试剂和收集阴道灌洗液和生殖道组织。这种小鼠模型将进一步阴道环境中的GBS宿主相互作用，这将导致潜在的治疗靶点怀孕期间母体控制阴道定植和避免传染给脆弱的新生儿的理解。它也将是感兴趣的，以增加在女性阴道我们的一般细菌 - 宿主相互作用的理解。

在这种模式的发展，多个观测结果就影响GBS定植阴道的时间因素使得。为了确定在接种的影响GBS细菌的持久性如何动情期，小鼠上演通过阴道灌洗液接种的一天。 图1示出的鼠标发情周期的四个阶段，由以下方法测定湿贴装阴道灌洗液，行之有效的方法29。小鼠分为基于该初始阶段组，并且经由阴道拭子随时间监测GBS持久性。在动情前期阶段接种的小?...

Watch this Scientific Journal Video about A Murine Model of Group B Streptococcus Vaginal Colonization at JoVE.com

A Murine Model of Group B Streptococcus Vaginal Colonization

此协议的目的是模仿人类的B组链球菌 （GBS）阴道定植在小鼠模型。此方法可用于研究宿主的免疫反应，并促进GBS阴道持久细菌因素，以及测试的治疗策略。

B组的小鼠模型<em&gt;链球菌</em&gt;阴道定植

Streptococcus agalactiae (group B Streptococcus, GBS), is a Gram-positive, asymptomatic colonizer of the human gastrointestinal tract and vaginal tract of ...

a-murine-model-of-group-b-streptococcus-vaginal-colonization

Research

JoVE Journal

Immunology and Infection

A Murine Model of Group B Streptococcus Vaginal Colonization

14.3K 次观看.  University of California San Diego School of Medicine. 此协议的目的是模仿人类的B组链球菌 （GBS）阴道定植在小鼠模型。此方法可用于研究宿主的免疫反应，并促进GBS阴道持久细菌因素，以及测试的治疗策略。 

视频: A Murine Model of Group B Streptococcus Vaginal Colonization

Protocols for Vaginal Inoculation and Sample Collection in the Experimental Mouse Model of Candida vaginitis

Vulvovaginal candidiasis (VVC), caused by Candida species, is a fungal infection of the lower female genital tract that affects approximately 75% of otherwise healthy women during their reproductive years18,32-34. Predisposing factors include antibiotic usage, uncontrolled diabetes and disturbance in reproductive hormone levels due to pregnancy, oral contraceptives or hormone replacement therapies33,34. Recurrent VVC (RVVC), defined as three or more episodes per year, affects a separate 5 to 8% of women with no predisposing factors33.
An experimental mouse model of VVC has been established and used to study the pathogenesis and mucosal host response to Candida3,4,11,16,17,19,21,25,37. This model has also been employed to test potential antifungal therapies in vivo13,24. The model requires that the animals be maintained in a state of pseudoestrus for optimal Candida colonization/infection6,14,23. Under such conditions, inoculated animals will have detectable vaginal fungal burden for weeks to months. Past studies show an extremely high parallel between the animal model and human infection relative to immunological and physiological properties3,16,21. Differences, however, include a lack of Candida as normal vaginal flora and a neutral vaginal pH in the mice.
Here, we demonstrate a series of key methods in the mouse vaginitis model that include vaginal inoculation, rapid collection of vaginal specimens, assessment of vaginal fungal burden, and tissue preparations for cellular extraction/isolation. This is followed by representative results for constituents of vaginal lavage fluid, fungal burden, and draining lymph node leukocyte yields. With the use of anesthetics, lavage samples can be collected at multiple time points on the same mice for longitudinal evaluation of infection/colonization. Furthermore, this model requires no immunosuppressive agents to initiate infection, allowing immunological studies under defined host conditions. Finally, the model and each technique introduced here could potentially give rise to use of the methodologies to examine other infectious diseases of the lower female genital tract (bacterial, parasitic, viral) and respective local or systemic host defenses.

Protocols for Vaginal Inoculation and Sample Collection in the Experimental Mouse Model of Candida vaginitis

Key techniques to be used in the evaluation of Candida vaginitis in an experimental animal model are described. The methods will allow rapid collection of vaginal specimens and lymphocytes from draining lumbar lymph nodes. These techniques could give rise to mouse models of other diseases in the female lower genital tract.

实验小鼠模型阴道接种和样品采集的协议念珠菌阴道炎

Vulvovaginal candidiasis (VVC), caused by Candida species, is a fungal infection of the lower female genital tract that affects approximately 75% of ...

科研

免疫与感染

Characterization of Inflammatory Responses During Intranasal Colonization with Streptococcus pneumoniae

Nasopharyngeal colonization by Streptococcus pneumoniae is a prerequisite to invasion to the lungs or bloodstream1. This organism is capable of colonizing the mucosal surface of the nasopharynx, where it can reside, multiply and eventually overcome host defences to invade to other tissues of the host. Establishment of an infection in the normally lower respiratory tract results in pneumonia. Alternatively, the bacteria can disseminate into the bloodstream causing bacteraemia, which is associated with high mortality rates2, or else lead directly to the development of pneumococcal meningitis. Understanding the kinetics of, and immune responses to, nasopharyngeal colonization is an important aspect of S. pneumoniae infection models.
Our mouse model of intranasal colonization is adapted from human models3 and has been used by multiple research groups in the study of host-pathogen responses in the nasopharynx4-7. In the first part of the model, we use a clinical isolate of S. pneumoniae to establish a self-limiting bacterial colonization that is similar to carriage events in human adults. The procedure detailed herein involves preparation of a bacterial inoculum, followed by the establishment of a colonization event through delivery of the inoculum via an intranasal route of administration. Resident macrophages are the predominant cell type in the nasopharynx during the steady state. Typically, there are few lymphocytes present in uninfected mice8, however mucosal colonization will lead to low- to high-grade inflammation (depending on the virulence of the bacterial species and strain) that will result in an immune response and the subsequent recruitment of host immune cells. These cells can be isolated by a lavage of the tracheal contents through the nares, and correlated to the density of colonization bacteria to better understand the kinetics of the infection.

Characterization of Inflammatory Responses During Intranasal Colonization with Streptococcus pneumoniae

Colonization of the murine nasopharynx with Streptococcus pneumoniae and the subsequent extraction of adherent or recruited cells is described. This technique involves flushing the nasopharynx and collection of the fluid through the nares and is adaptable for various readouts, including differential cell quantification and analysis of mRNA expression in situ.

肺炎链球菌在鼻内殖民期间炎症反应的特征

Nasopharyngeal colonization by Streptococcus pneumoniae is a prerequisite to invasion to the lungs or bloodstream1. This organism is capable of colonizing ...

Humanized Mouse Model to Study Bacterial Infections Targeting the Microvasculature

Neisseria meningitidis causes a severe, frequently fatal sepsis when it enters the human blood stream. Infection leads to extensive damage of the blood vessels resulting in vascular leak, the development of purpuric rashes and eventual tissue necrosis. Studying the pathogenesis of this infection was previously limited by the human specificity of the bacteria, which makes in vivo models difficult. In this protocol, we describe a humanized model for this infection in which human skin, containing dermal microvessels, is grafted onto immunocompromised mice. These vessels anastomose with the mouse circulation while maintaining their human characteristics. Once introduced into this model, N. meningitidis adhere exclusively to the human vessels, resulting in extensive vascular damage, inflammation and in some cases the development of purpuric rash. This protocol describes the grafting, infection and evaluation steps of this model in the context of N. meningitidis infection. The technique may be applied to numerous human specific pathogens that infect the blood stream.

Neisseria meningitidis is a human specific pathogen that infects blood vessels. In this protocol human microvessels are introduced into a mouse by grafting human skin onto immunocompromised mice. Bacteria adhere extensively to the human vessels, leading to vascular damage and development of the purpuric rash typically observed in human cases.

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 ...

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

Chronic inflammation is a major driver of pathological tissue damage and a unifying characteristic of many chronic diseases in humans including neoplastic, autoimmune, and chronic inflammatory diseases. Emerging evidence implicates pathogen-induced chronic inflammation in the development and progression of chronic diseases with a wide variety of clinical manifestations. Due to the complex and multifactorial etiology of chronic disease, designing experiments for proof of causality and the establishment of mechanistic links is nearly impossible in humans. An advantage of using animal models is that both genetic and environmental factors that may influence the course of a particular disease can be controlled. Thus, designing relevant animal models of infection represents a key step in identifying host and pathogen specific mechanisms that contribute to chronic inflammation.
Here we describe a mouse model of pathogen-induced chronic inflammation at local and systemic sites following infection with the oral pathogen Porphyromonas gingivalis, a bacterium closely associated with human periodontal disease. Oral infection of specific-pathogen free mice induces a local inflammatory response resulting in destruction of tooth supporting alveolar bone, a hallmark of periodontal disease. In an established mouse model of atherosclerosis, infection with P. gingivalis accelerates inflammatory plaque deposition within the aortic sinus and innominate artery, accompanied by activation of the vascular endothelium, an increased immune cell infiltrate, and elevated expression of inflammatory mediators within lesions. We detail methodologies for the assessment of inflammation at local and systemic sites. The use of transgenic mice and defined bacterial mutants makes this model particularly suitable for identifying both host and microbial factors involved in the initiation, progression, and outcome of disease. Additionally, the model can be used to screen for novel therapeutic strategies, including vaccination and pharmacological intervention.

Animal models have proven to be invaluable tools in defining host and pathogen specific mechanisms that contribute to the development of chronic inflammation. Here we describe a mouse model of oral infection with the human pathogen Porphyromonas gingivalis and detail methodologies to assess the progression of 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 ...

Non-Invasive Model of Neuropathogenic Escherichia coli Infection in the Neonatal Rat

Investigation of the interactions between animal host and bacterial pathogen is only meaningful if the infection model employed replicates the principal features of the natural infection. This protocol describes procedures for the establishment and evaluation of systemic infection due to neuropathogenic Escherichia coli K1 in the neonatal rat. Colonization of the gastrointestinal tract leads to dissemination of the pathogen along the gut-lymph-blood-brain course of infection and the model displays strong age dependency. A strain of E. coli O18:K1 with enhanced virulence for the neonatal rat produces exceptionally high rates of colonization, translocation to the blood compartment and invasion of the meninges following transit through the choroid plexus. As in the human host, penetration of the central nervous system is accompanied by local inflammation and an invariably lethal outcome. The model is of proven utility for studies of the mechanism of pathogenesis, for evaluation of therapeutic interventions and for assessment of bacterial virulence.

Non-Invasive Model of Neuropathogenic Escherichia coli Infection in the Neonatal Rat

Here, a procedure is described for the establishment of systemic infection in the neonatal rat with cultures of Escherichia coli K1. This non-invasive procedure permits colonization of the gastrointestinal tract, translocation of the pathogen to the systemic circulation, and invasion of the central nervous system at the choroid plexus.

Neuropathogenic的非侵入型<em&gt;大肠杆菌</em&gt;感染乳鼠

Investigation of the interactions between animal host and bacterial pathogen is only meaningful if the infection model employed replicates the principal ...

Recurrent Escherichia coli Urinary Tract Infection Triggered by Gardnerella vaginalis Bladder Exposure in Mice

Recurrent urinary tract infections (rUTI) caused by uropathogenic Escherichia coli (UPEC) are common and costly. Previous articles describing models of UTI in male and female mice have illustrated the procedures for bacterial inoculation and enumeration in urine and tissues. During an initial bladder infection in C57BL/6 mice, UPEC establish latent reservoirs inside bladder epithelial cells that persist following clearance of UPEC bacteriuria. This model builds on these studies to examine rUTI caused by the emergence of UPEC from within latent bladder reservoirs. The urogenital bacterium Gardnerella vaginalis is used as the trigger of rUTI in this model because it is frequently present in the urogenital tracts of women, especially in the context of vaginal dysbiosis that has been associated with UTI. In addition, a method for in situ bladder fixation followed by scanning electron microscopy (SEM) analysis of bladder tissue is also described, with potential application to other studies involving the bladder.

Recurrent Escherichia coli Urinary Tract Infection Triggered by Gardnerella vaginalis Bladder Exposure in Mice

A mouse model of uropathogenic E. coli (UPEC) transurethral inoculation to establish latent intracellular bladder reservoirs and subsequent bladder exposure to G. vaginalis to induce recurrent UPEC UTI is demonstrated. Also demonstrated are the enumeration of bacteria, urine cytology, and in situ bladder fixation and processing for scanning electron microscopy.

由加德纳菌阴道膀胱暴露在小鼠中引发的复发性大肠杆菌尿路感染

Recurrent urinary tract infections (rUTI) caused by uropathogenic Escherichia coli (UPEC) are common and costly. Previous articles describing models of ...

C-section of Preclinical Animal Model of Chorioamnionitis Triggered by Group B Streptococcus (GBS)

Group B Streptococcus (GBS) is one of the most common bacteria isolated during human pregnancy. It is a leading cause of placental infection/inflammation, termed chorioamnionitis. Chorioamnionitis exposes the developing fetus to a high risk of organ injuries, perinatal morbidity, and mortality, as well as life-long neurobehavioral impairments and other non-neurological developmental issues. The two most frequent subtypes of GBS isolates from maternal and fetal tissues are serotypes Ia (13%-23%) and III (25%-53%). Our lab has developed and characterized a rat model of GBS-induced chorioamnionitis to study subsequent impacts on the central nervous system of the developing fetus and to understand underlying mechanistic aspects. This article presents the design as well as uses of the preclinical rat model, which closely reproduces the hallmark of GBS-induced chorioamnionitis in humans. This article aims to help scientists reproduce the experimental design as well as to provide support through examples of troubleshooting. The present model may also contribute to potential discoveries through uncovering causes, mechanisms, and novel therapeutic avenues, which remain unsettled in many developmental impairments arising from chorioamnionitis. Furthermore, the use of this model may be extended to the studies of perinatal non-neurological common and severe morbidities affecting, for instance, the retina, bowel, lung, and kidney. The main interest of this research is in the field of GBS-induced fetal neurodevelopmental impairments such as cerebral palsy (CP), attention deficit hyperactivity disorder (ADHD), and autism spectrum disorder (ASD). The rationale supporting this model is presented in this article, followed by procedures and results.

<meta charset="utf8"></head><body>B 组链球菌 GBS 引发的绒毛膜羊膜炎临床前动物模型的剖腹产

The goal of this protocol is to describe a preclinical animal model of Group B Streptococcus (GBS)-induced chorioamnionitis. The study is designed to investigate mechanistic processes, potential causal links with developmental impairments, and finally to develop translational anti-inflammatory placento- and neuro-protective treatments.

B 组 链球菌 （GBS） 引发的绒毛膜羊膜炎临床前动物模型的剖腹产

Group B Streptococcus (GBS) is one of the most common bacteria isolated during human pregnancy. It is a leading cause of placental infection/inflammation, ...

用于研究阴道微生物组的微流体阴道芯片

Modeling Healthy and Dysbiotic Vaginal Microenvironments in a Human Vagina-on-a-Chip

Women's health, and particularly diseases of the female reproductive tract (FRT), have not received the attention they deserve, even though an unhealthy reproductive system may lead to life-threatening diseases, infertility, or adverse outcomes during pregnancy. One barrier in the field is that there has been a dearth of preclinical, experimental models that faithfully mimic the physiology and pathophysiology of the FRT. Current in vitro and animal models do not fully recapitulate the hormonal changes, microaerobic conditions, and interactions with the vaginal microbiome. The advent of Organ-on-a-Chip (Organ Chip) microfluidic culture technology that can mimic tissue-tissue interfaces, vascular perfusion, interstitial fluid flows, and the physical microenvironment of a major subunit of human organs can potentially serve as a solution to this problem. Recently, a human Vagina Chip that supports co-culture of human vaginal microbial consortia with primary human vaginal epithelium that is also interfaced with vaginal stroma and experiences dynamic fluid flow has been developed. This chip replicates the physiological responses of the human vagina to healthy and dysbiotic microbiomes. A detailed protocol for creating human Vagina Chips has been described in this article.

作者聚焦：使用阴道芯片彻底改变阴道微生物组相互作用的研究

This article describes a protocol for creating a microfluidic vagina-on-a-chip (Vagina Chip) culture device that enables the study of human host interactions with a living vaginal microbiome under microaerophilic conditions. This chip can be used as a tool to investigate vaginal diseases as well as to develop and test potential therapeutic countermeasures.

在人类阴道芯片中模拟健康和生态失调的阴道微环境

Women's health, and particularly diseases of the female reproductive tract (FRT), have not received the attention they deserve, even though an unhealthy ...

B组的小鼠模型

摘要

探索更多视频

实验小鼠模型阴道接种和样品采集的协议念珠菌阴道炎

肺炎链球菌在鼻内殖民期间炎症反应的特征

人源化的小鼠模型来研究细菌感染瞄准微血管

鼠标型号病原体引起的慢性炎症局部及全身站点

Neuropathogenic的非侵入型

由加德纳菌阴道膀胱暴露在小鼠中引发的复发性大肠杆菌尿路感染

B 组链球菌（GBS）引发的绒毛膜羊膜炎临床前动物模型的剖腹产

B 组链球菌（GBS）引发的绒毛膜羊膜炎临床前动物模型的剖腹产

在人类阴道芯片中模拟健康和生态失调的阴道微环境

在人类阴道芯片中模拟健康和生态失调的阴道微环境

B组的小鼠模型

摘要

探索更多视频

实验小鼠模型阴道接种和样品采集的协议念珠菌阴道炎

肺炎链球菌在鼻内殖民期间炎症反应的特征

人源化的小鼠模型来研究细菌感染瞄准微血管

鼠标型号病原体引起的慢性炎症局部及全身站点

Neuropathogenic的非侵入型

由加德纳菌阴道膀胱暴露在小鼠中引发的复发性大肠杆菌尿路感染

B 组 链球菌 （GBS） 引发的绒毛膜羊膜炎临床前动物模型的剖腹产

B 组 链球菌 （GBS） 引发的绒毛膜羊膜炎临床前动物模型的剖腹产

在人类阴道芯片中模拟健康和生态失调的阴道微环境

在人类阴道芯片中模拟健康和生态失调的阴道微环境

B 组链球菌（GBS）引发的绒毛膜羊膜炎临床前动物模型的剖腹产

B 组链球菌（GBS）引发的绒毛膜羊膜炎临床前动物模型的剖腹产