Name: intracranial subarachnoidal route of infection for investigating roles of streptococcus suis biofilms in meningitis in a mouse infection model
Uploaded: 2018-07-01T13:00:00.000+00:00
Duration: 7 min 26 s
Description: Watch this Scientific Journal Video about Intracranial Subarachnoidal Route of Infection for Investigating Roles of Streptococcus suis Biofilms in Meningitis in a Mouse Infection Model at JoVE.com

这项工作得到了中国国家重点研究开发计划 (2017YFD0500102) 的资助;中国国家自然科学基金 [31572544];兽医病因生物学国家重点实验室 [SKLVEB2016KFKT005];上海农业应用技术发展项目, 中国 [G2016060201]。

小鼠感染实验由中国江苏省实验动物监测委员会批准, 并在南京农业大学实验动物中心 (许可证编号: SYXK (Su) 2017-0007) 上进行。
1. 细菌的制备
注:猪血清2型毒株 P1/7 是从患病的猪与脑膜炎19分离出来的。P1/7 菌株生长在托德-休伊特汤 (THB, 公式每公升 THB: 心脏输液, 3.1 克; neopeptone, 20.0 克; 葡萄糖, 2.0 克; 氯化钠, 2.0 克; 磷酸二钠, 0.4 克; 碳酸钠, 2.5 克) 和电镀在托德-威特特琼脂 (临屋区, 公式每公升心脏输液, 3.1 克;neopeptone, 20.0 克;葡萄糖, 2.0 克;氯化钠, 2.0 克;磷酸二钠, 0.4 克;碳酸钠, 2.5 克;琼脂, 15.0 克) 在37°c 和 5% CO2。
<ol><li>应变 P1/7 浮游细胞的采集<ol>
<li>收集5毫升浮游细胞从中对数阶段培养 (OD600= 0.6), 离心机为3分钟在 8000 x g, 然后洗涤3次与 PBS。</li>
		<li>并用重悬的细胞与5毫升25% 甘油在 THB, 整除成5管, 然后存储在-80 摄氏度。</li>
	</ol>
</li>
	<li>应变 P1/7 生物膜状态细胞的采集<ol>
<li>采取20毫升隔夜文化和增加180毫升新鲜的 THB;将稀释过的文化分成10圆的培养板, 然后把盘子放在37摄氏度的孵化器中, 5% CO2为 24 h。</li>
		<li>孵化后, 轻轻摇动盘子, 并用重悬不粘在盘子上的细菌, 然后用吸入法丢弃上清。 注意: 轻轻摇动盘子, 避免 resuspending 沉淀物。</li>
		<li>加入5毫升的 PBS, 以收获生物膜状态细胞, 完全并用重悬沉积物, 然后将样品转移到新管上。</li>
		<li>油脂实验生物膜状态细胞从最后一步以以下参量:60 W, 4 个周期, 5 s 在和十年代。</li>
		<li>离心机为3分钟在 8000 x g 和存放上清在-80 °c。 注: 上清可能含有生物膜成分, 可用于在感染前并用重悬生物膜细菌, 如动物实验所述 (步骤 3)。</li>
		<li>在 THB 中并用重悬10毫升25% 甘油的沉淀物, 然后将生物膜状态细胞储存在-80 摄氏度。</li>
	</ol>
</li>
</ol>2. 扫描电镜 (SEM) 分析
<ol><li>用4% 多聚甲醛 12-24 小时在4摄氏度固定收集的生物膜状态细胞和浮游细胞。</li>
	<li>用蒸馏水快速冲洗样品, 并以乙醇 (25%、50%、70%、90% 和 100%) 的浓度增加为30分钟, 在每个溶液中依次脱水样品。</li>
	<li>将干试样粘附在带有双面胶带的金属夹上, 最后将其涂在带有金和钯的蒸发器中。</li>
	<li>用扫描电子显微镜观察样品, 使用以下参数: EHT (超高压) = 10 伏;WD (工作距离) = 7.0 或8.0 毫米;放大 = 5000 x;信号 A = SE1。</li>
</ol>3. 动物实验
<ol><li>在本研究中使用 SPF 6 周的雌性 BALB/c 小鼠。通过颅内 subarachnoidal 感染的所有小鼠, 其注射部位位于 bregma 3.5 毫米延髓。Chiavolini 和al17清楚地描述了注射部位的立体定向坐标。<ol>
<li>病理学分析, 感染两组小鼠 (每组5只小鼠), 使用浮游细胞或生物膜状态细胞, 剂量为 3 x 107菌落形成单位 (CFU), 另有5只老鼠注射 PBS 作为控制。</li>
		<li>为检测 TLR2 和细胞因子在脑中的 mRNA 表达, 在 3 x 107 CFU 中使用浮游细胞或生物膜状态细胞感染两组小鼠 (每组6只小鼠)。</li>
		<li>通过电镀连续稀释来确定活菌的数量。</li>
		<li>弄死所有小鼠在12小时后感染后进行进一步的研究。 注: 在感染前必须清除在培养基中的甘油。从-80 °c 恢复的浮游细胞和生物膜状态细胞被洗涤3次与 PBS。然后, 浮游细胞悬浮与 PBS, 稀释到适当剂量的感染;生物膜状态细胞与储存的上清液悬浮 (从步 1.2.5) 和稀释到适当剂量的感染。感染量不应超过50µL。</li>
	</ol>
</li>
	<li>脑组织中 TLR2 和细胞因子 mRNA 表达的检测<ol>
<li>使用 rna 提取试剂盒从脑组织中提取总 RNA。<ol>
<li>对于每只老鼠, 使用全脑组织提取 RNA。把整个脑组织分成5管。服用多达100毫克的脑组织, 并添加1毫升的裂解液, 每管含有株溶藻基质提供的试剂盒。</li>
			<li>在6.0 的设置中, 在一个均质机上处理管, 然后离心管在 12000 x g 和4°c 为5分钟。</li>
			<li>离心后, 将上相转移到一个新的离心管。</li>
			<li>在室温下孵化转移样品5分钟;添加300µL 的氯仿, 漩涡十年代, 然后孵化5分钟室温。</li>
			<li>离心管在 12000 x g 和4°c 5 分钟转移上部阶段到新的管子。</li>
			<li>将500µL 的冷绝对乙醇添加到管中, 然后将其反转5次, 然后将样品贮存在-20 摄氏度, 至少为1小时。</li>
			<li>离心管在 12000 x g 和4°c 为15分钟和去除上清。用500µL 冷75% 乙醇在 RNase 的 H2O 上清洗颗粒。</li>
			<li>除去乙醇, 空气干燥的颗粒在室温下5分钟, 并并用重悬 RNA 在100µL RNase 无2O。</li>
			<li>在室温下孵化 rna 5 分钟, 用 rna bioanalyzer 测定 rna 浓度。</li>
		</ol>
</li>
		<li>进行 cDNA 合成, 包括消除基因组 DNA (gDNA), 使用 thermocycler 与反向转录 (RT) 试剂盒。<ol>
<li>将1µg 的 RNA 添加到含有2µL 5x gDNA 消除缓冲液的管中, 1 µL gDNA 消除酶和 RNase 的 H2O, 直到体积达到10µL. 在2摄氏度孵育42分钟。</li>
			<li>添加10µL 的主混合 (1 µL 的 rt 酶混合 I, 1 µL 的 rt 底漆混合, 4 µL 5x 缓冲器 2, 4 µL RNase 免费 H2O) 到反应解决方案从最后一步, 然后轻轻混合。立即进行 RT 反应:37 °c 为15分钟, 然后85°c 5 s。</li>
			<li>使用 SYBR qPCR 试剂盒实时 pcr 机进行定量实时 pcr (RT qPCR) 分析。添加10µL 的2x 酶, 0.8 µL 的前底漆, 0.8 µL 的反向底漆, 0.4 µL 50x 火箭参考染料 II, 2 µL 的 cDNA 模板, 和 RNase 免费 H2O, 直到总容量20µL。</li>
			<li>使用以下热参数运行每个样本: 三十年代在94°c, 跟随40个周期 5 s 在95°c 和三十四年代在60°c, 以十五年代的最后阶段在95°c, 1 分钟在60°c, 十五年代在95摄氏度。qPCR 的底漆列于表 1。管家基因B2m和18s rRNA被用作内部控制。</li>
		</ol>
</li>
		<li>根据 2-ΔΔCt方法计算相对褶皱变化。</li>
	</ol>
</li>
	<li>
组织学切片观察
	<ol>
<li>24 h 固定10% 福尔马林后, 将固定的脑组织重建成适当大小的组织块, 在 2-3 毫米。</li>
		<li>把纸巾放在嵌入的卡带里, 浸泡在一个新的固定液中, 用自动真空脱水脱水。</li>
		<li>取出固定组织, 并在每种溶液中依次脱水以增加乙醇 (70%、80%、95%、95% 和 100%) 的浓度。</li>
		<li>将该样品与二甲苯 Transparentize 15 分钟, 将其浸在液体石蜡中60°c 90 分钟, 然后用嵌入机嵌入。</li>
		<li>将石蜡块嵌入组织切成3µm 片, 在45摄氏度的水上平铺切片, 然后在空气中晾干。</li>
		<li>在60摄氏度孵育3小时的切片, 用 haematoxylin 和伊红 (他) 方法染色, 并用中性胶封住。</li>
		<li>用光学显微镜观察大脑的组织病理学变化。采用四点分级系统评价组织学变化。使用未配对的t检验进行统计分析。 充血/出血:缺席, 评分 0;小面积的局灶充血/出血, 评分 1;大面积的局灶充血/出血, 或多处小面积的局灶充血/出血, 评分 2;多达三个大焦距充血/出血部位, 得分 3;弥漫性充血/出血, 评分4。 坏死:缺席, 评分 0;病灶坏死, 评分 1;弥漫性发生 1-10 坏死细胞, 评分 2;汇合坏死灶, 评分 3;广泛的汇合坏死, 评分4。 炎症细胞浸润:缺席, 评分 0;少数和焦点渗透, 得分 1;多焦浸润或渗透形成聚集, 得分 2;最多三集, 分数 3;超过四集, 分数4。</li>
	</ol>
</li>
</ol>

<ol>
	<li>Gottschalk, M., Xu, J., Calzas, C., Segura, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Streptococcus+suis:+a+new+emerging+or+an+old+neglected+zoonotic+pathogen?.">Streptococcus suis: a new emerging or an old neglected zoonotic pathogen?.</a> Future Microbiology. 5, 371-391 (2010).</li><li>Goyette-Desjardins, G., Auger, J. P., Xu, J., Segura, M., Gottschalk, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Streptococcus+suis,+an+important+pig+pathogen+and+emerging+zoonotic+agent-an+update+on+the+worldwide+distribution+based+on+serotyping+and+sequence+typing.">Streptococcus suis, an important pig pathogen and emerging zoonotic agent-an update on the worldwide distribution based on serotyping and sequence typing.</a> Emerging Microbes &amp; Infections. 3 (6), e45 (2014).</li><li>Kerdsin, 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=Emergence+of+Streptococcus+suis+serotype+9+infection+in+humans.">Emergence of Streptococcus suis serotype 9 infection in humans.</a> Journal of Microbiology, Immunology and Infection. 50 (4), 545-546 (2017).</li><li>Hatrongjit, 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=First+human+case+report+of+sepsis+due+to+infection+with+Streptococcus+suis+serotype+31+in+Thailand.">First human case report of sepsis due to infection with Streptococcus suis serotype 31 in Thailand.</a> BMC Infect Diseases. 15, 392 (2015).</li><li>Fittipaldi, N., Segura, M., Grenier, D., Gottschalk, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Virulence+factors+involved+in+the+pathogenesis+of+the+infection+caused+by+the+swine+pathogen+and+zoonotic+agent+Streptococcus+suis.">Virulence factors involved in the pathogenesis of the infection caused by the swine pathogen and zoonotic agent Streptococcus suis.</a> Future Microbiology. 7 (2), 259-279 (2012).</li><li>Hall-Stoodley, L., Costerton, J. W., Stoodley, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bacterial+biofilms:+from+the+natural+environment+to+infectious+diseases.">Bacterial biofilms: from the natural environment to infectious diseases.</a> Nature Reviews Microbiology. 2 (2), 95-108 (2004).</li><li>Fuxman Bass, J. 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=Extracellular+DNA:+a+major+proinflammatory+component+of+Pseudomonas+aeruginosa+biofilms.">Extracellular DNA: a major proinflammatory component of Pseudomonas aeruginosa biofilms.</a> Journal of Immunology. 184 (11), 6386-6395 (2010).</li><li>Isiaku, A. 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=Biofilm+is+associated+with+chronic+streptococcal+meningoencephalitis+in+fish.">Biofilm is associated with chronic streptococcal meningoencephalitis in fish.</a> Microbial Pathogenesis. 102, 59-68 (2017).</li><li>Oggioni, M. 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=Switch+from+planktonic+to+sessile+life:+a+major+event+in+pneumococcal+pathogenesis.">Switch from planktonic to sessile life: a major event in pneumococcal pathogenesis.</a> Molecular Microbiology. 61 (5), 1196-1210 (2006).</li><li>Xiao, 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=Streptococcus+suis+small+RNA+rss04+contributes+to+the+induction+of+meningitis+by+regulating+capsule+synthesis+and+by+inducing+biofilm+formation+in+a+mouse+infection+model.">Streptococcus suis small RNA rss04 contributes to the induction of meningitis by regulating capsule synthesis and by inducing biofilm formation in a mouse infection model.</a> Veterinary Microbiology. 199, 111-119 (2017).</li><li>Dominguez-Punaro, M. 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=Streptococcus+suis+serotype+2,+an+important+swine+and+human+pathogen,+induces+strong+systemic+and+cerebral+inflammatory+responses+in+a+mouse+model+of+infection.">Streptococcus suis serotype 2, an important swine and human pathogen, induces strong systemic and cerebral inflammatory responses in a mouse model of infection.</a> Journal of Immunology. 179 (3), 1842-1854 (2007).</li><li>Seitz, 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=A+novel+intranasal+mouse+model+for+mucosal+colonization+by+Streptococcus+suis+serotype+2.">A novel intranasal mouse model for mucosal colonization by Streptococcus suis serotype 2.</a> Journal of Medical Microbiology. 61 (Pt 9), 1311-1318 (2012).</li><li>Busque, P., Higgins, R., Caya, F., Quessy, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Immunization+of+pigs+against+Streptococcus+suis+serotype+2+infection+using+a+live+avirulent+strain.">Immunization of pigs against Streptococcus suis serotype 2 infection using a live avirulent strain.</a> Canadian Journal of Veterinary Research. 61 (4), 275-279 (1997).</li><li>Williams, A. E., Blakemore, W. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pathology+of+Streptococcal+meningitis+following+intravenous+intracisternal+and+natural+routes+of+infection.">Pathology of Streptococcal meningitis following intravenous intracisternal and natural routes of infection.</a> Neuropathology and Applied Neurobiology. 4 (4), 345-356 (1990).</li><li>Dominguez-Punaro, M. 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=Severe+cochlear+inflammation+and+vestibular+syndrome+in+an+experimental+model+of+Streptococcus+suis+infection+in+mice.">Severe cochlear inflammation and vestibular syndrome in an experimental model of Streptococcus suis infection in mice.</a> European Journal of Clinical Microbiology &amp; Infectious Diseases. 31 (9), 2391-2400 (2012).</li><li>Auger, J. P., Fittipaldi, N., Benoit-Biancamano, M. O., Segura, M., Gottschalk, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Virulence+Studies+of+Different+Sequence+Types+and+Geographical+Origins+of+Streptococcus+suis+Serotype+2+in+a+Mouse+Model+of+Infection.">Virulence Studies of Different Sequence Types and Geographical Origins of Streptococcus suis Serotype 2 in a Mouse Model of Infection.</a> Pathogens. 5 (3), (2016).</li><li>Chiavolini, 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=Method+for+inducing+experimental+pneumococcal+meningitis+in+outbred+mice.">Method for inducing experimental pneumococcal meningitis in outbred mice.</a> BMC Microbiology. 4, 36 (2004).</li><li>Madsen, L. W., Svensmark, B., Elvestad, K., Jensen, H. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Otitis+interna+is+a+frequent+sequela+to+Streptococcus+suis+meningitis+in+pigs.">Otitis interna is a frequent sequela to Streptococcus suis meningitis in pigs.</a> Veterinary Pathology. 38 (2), 190-195 (2001).</li><li>Holden, M. T., 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=Rapid+evolution+of+virulence+and+drug+resistance+in+the+emerging+zoonotic+pathogen+Streptococcus+suis.">Rapid evolution of virulence and drug resistance in the emerging zoonotic pathogen Streptococcus suis.</a> PLoS One. 4 (7), e6072 (2009).</li><li>Dominguez-Punaro Mde, 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=In+vitro+characterization+of+the+microglial+inflammatory+response+to+Streptococcus+suis,+an+important+emerging+zoonotic+agent+of+meningitis.">In vitro characterization of the microglial inflammatory response to Streptococcus suis, an important emerging zoonotic agent of meningitis.</a> Infection and Immunity. 78 (12), 5074-5085 (2010).</li><li>Hassan, 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=Evaluation+of+different+detection+methods+of+biofilm+formation+in+the+clinical+isolates.">Evaluation of different detection methods of biofilm formation in the clinical isolates.</a> Brazilian Journal of Infectious Diseases. 15 (4), 305-311 (2011).</li><li>Vanier, 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=New+putative+virulence+factors+of+Streptococcus+suis+involved+in+invasion+of+porcine+brain+microvascular+endothelial+cells.">New putative virulence factors of Streptococcus suis involved in invasion of porcine brain microvascular endothelial cells.</a> Microbial Pathogenesis. 46 (1), 13-20 (2009).</li><li>Takeuchi, 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=The+contribution+of+suilysin+to+the+pathogenesis+of+Streptococcus+suis+meningitis.">The contribution of suilysin to the pathogenesis of Streptococcus suis meningitis.</a> Journal of Infectious Diseases. 209 (10), 1509-1519 (2014).</li><li>Tenenbaum, T., 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=Polar+bacterial+invasion+and+translocation+of+Streptococcus+suis+across+the+blood-cerebrospinal+fluid+barrier+in+vitro.">Polar bacterial invasion and translocation of Streptococcus suis across the blood-cerebrospinal fluid barrier in vitro.</a> Cellular Microbiology. 11 (2), 323-336 (2009).</li></ol>

作者没有什么可透露的。

这里描述的颅内 subarachnoidal 感染途径与其他感染途径相比有明显的优势。它允许调查人员研究宿主-细菌相互作用和细菌成分对宿主免疫应答的影响直接在大脑, 这模仿细菌进入中枢神经系统。因此, 这种感染途径可以扩展, 以调查其他细菌引起的脑膜炎机制。此外, 还可用于检测药物对细菌性脑膜炎的疗效。
为了获得良好的结果, 使用这个模型, 以下关键步骤是明确的。生物膜...

猪链球菌(猪) 是全世界猪的主要细菌病原体, 造成严重疾病, 包括脑膜炎、肺炎、败血症、心内膜炎和关节炎1。它也是一个新兴的人畜共患病的特工。到目前为止, 据报道, 九型血清型可能导致人类感染, 包括血清型 2, 4, 5, 9, 14, 16, 21, 24, 312,3,4。在人类和猪中, 脑膜炎是感染猪的主要临床征象之一。在越南和泰国,美国猪是5成人脑膜炎的主要病因。微生物生物膜是粘在一起的微生物, 并集中在界面上;它们对细菌的毒力、不同环境下的生存和耐药性5至关重要。生物膜通常被一个细胞外基质所包围, 通常含有多糖、蛋白质和 DNA6。后者能够诱发宿主炎症反应和细胞因子的产生7。在以前的研究中, 生物膜的形成已被用于链球菌脑膜炎。生物膜有助于罗非鱼鱼模型中的链球菌脑膜炎链球菌, 生物膜的形成已经通过腹腔内接种8在体内的脑组织和脑膜表面周围显露出来。在脑膜炎期间,肺炎链球菌处于生物膜状状态, 而这种生物膜状态下的细菌更有效地诱发了老鼠感染模型9中的脑膜炎。此外, 在我们以前的研究中, 在小鼠脑中与猪的相关的生物膜状态通过生存分析导致细菌毒力10。然而, 直接证据的生物膜参与的猪链球菌脑膜炎需要进一步的调查。
采用腹腔 (ip)11、鼻腔 (i.n.)12、静脉注射 (静脉注射)13和 intracisternal (中) 感染14条的方法, 研制出了猪鼠感染动物模型.15,16. 然而, 感染的 ip、i.n. 和静脉注射途径不适合于直接在脑部研究在脑膜炎中猪的表面成分的作用。这些包括生物膜胞外基质。虽然在美国猪感染中使用了内源接种, 但是这些文件中没有描述精确的注射部位。与此相反, 颅内 subarachnoidal 接种的注射部位的立体定向坐标在前一项研究中已经明确描述过17。这允许容易地承认接种点和更加简单的实验性协议。此外, 颅内 subarachnoidal 感染的途径, 模拟细菌进入中枢神经系统从鼻窦或中耳17, 中耳和脑膜炎的关系, 由猪.已经证明了马德森et al18。此外, 通过应用颅内 subarachnoidal 感染的小鼠, 我们已经证明了猪的小 RNA rss04 导致脑膜炎在我们以前的研究10。
在本研究中, 采用颅内 subarachnoidal 感染途径对小鼠进行了研究, 探讨了生物膜在猪链球菌脑膜炎中的作用。通过这种感染途径, 小鼠感染了浮游细胞或猪的生物膜状态细胞。生物膜对小鼠脑组织中 TLR2 和细胞因子表达的组织病理学分析清楚地表明,猪的生物膜对脑膜炎有促进作用。

<table><tbody><tr><td>Todd Hewitt Broth(THB)</td><td>Becton, Dickinson and Company</td><td>DF0492078</td><td>Dissolve 30 g of the powder in 1 L of purified water. Autoclave at 121&amp;deg; for 15 min.</td></tr><tr><td>Agar</td><td>DSBIO</td><td>16C0050</td><td>Dissolve 15 g of the powder in 1 L of THB. Autoclave at 121&amp;deg; for 15 min.</td></tr><tr><td>Milli-Q Reference Water Purification System</td><td>Merck KGaA</td><td>Z00QSVCUS</td><td>Without Dnase/ Rnase</td></tr><tr><td>NaCl</td><td>Tianjin Kemiou Chemical Reagent Co., Ltd</td><td>10019318</td><td>Dissolve 8 g NaCl, 0.2 g KCl, 1.42 g Na2HPO3 , 0.27 g KH2PO4 in 1 L of purified water. Autoclave at 121&amp;deg; for 15 min. Use KOH to adjust pH to 7.4.</td></tr><tr><td>Na2HPO3</td><td>Xilong Scientific Co., Ltd</td><td>9009012-01-09</td><td>Dissolve 8 g NaCl, 0.2 g KCl, 1.42 g Na2HPO3 , 0.27 g KH2PO4 in 1 L of purified water. Autoclave at 121&amp;deg; for 15 min. Use KOH to adjust pH to 7.4.</td></tr><tr><td>KCl</td><td>Xilong Scientific Co., Ltd</td><td>9009017-01-09</td><td>Dissolve 8 g NaCl, 0.2 g KCl, 1.42 g Na2HPO3 , 0.27 g KH2PO4 in 1 L of purified water. Autoclave at 121&amp;deg; for 15 min. Use KOH to adjust pH to 7.4.</td></tr><tr><td>KH2PO4</td><td>Xilong Scientific Co., Ltd</td><td>9009019-01-09</td><td>Dissolve 8 g NaCl, 0.2 g KCl, 1.42 g Na2HPO3 , 0.27 g KH2PO4 in 1 L of purified water. Autoclave at 121&amp;deg; for 15 min. Use KOH to adjust pH to 7.4.</td></tr><tr><td>KOH</td><td>Xilong Scientific Co., Ltd</td><td>9009014-01-09</td><td>Dissolve 8 g NaCl, 0.2 g KCl, 1.42 g Na2HPO3 , 0.27 g KH2PO4 in 1 L of purified water. Autoclave at 121&amp;deg; for 15 min. Use KOH to adjust pH to 7.4.</td></tr><tr><td>Glycerol</td><td>Sionpharm Chemical Reagent Co., Ltd</td><td>10010618</td><td>Diluted with equal volumu of purified water, autoclave at 121&amp;deg; for 15 min</td></tr><tr><td>4% paraformaldehyde</td><td>Sionpharm Chemical Reagent Co., Ltd</td><td>80096675</td><td></td></tr><tr><td>25% Glutaraldehyde</td><td>Sionpharm Chemical Reagent Co., Ltd</td><td>30092436</td><td>10-fold diluted with purified water for fixation.</td></tr><tr><td>Ethanol</td><td>Sionpharm Chemical Reagent Co., Ltd</td><td>10009218</td><td></td></tr><tr><td>Chloroform</td><td>Sionpharm Chemical Reagent Co., Ltd</td><td>10006818</td><td></td></tr><tr><td>Spctrophotometre</td><td>DeNovix Inc.</td><td>DS-11+</td><td></td></tr><tr><td>Ultrasound cell crusher</td><td>NingBo Scientz Biotechnology Co.,Ltd</td><td>JY96-IIN</td><td></td></tr><tr><td>Centrifuge</td><td>Hitachi Koki Co., Ltd</td><td>CT15RE</td><td></td></tr><tr><td>Refrigerator</td><td>Aucma Co., Ltd</td><td>DW-86L500</td><td></td></tr><tr><td>Scanning electron microscope</td><td>Zeiss</td><td>EVO-LS10</td><td></td></tr><tr><td>FastRNA Pro Green Kit</td><td>MP Biomedicals</td><td>#6045-050</td><td></td></tr><tr><td>FastPrep-24 Instrument</td><td>MP Biomedicals</td><td>116005500</td><td></td></tr><tr><td>Instrument for PCR</td><td>SensoQuest GmbH</td><td>1124310110</td><td></td></tr><tr><td>QuantStudio 6 Flex</td><td>Thermo Fisher Scientific</td><td>4485689</td><td></td></tr><tr><td>SYBR Premix Ex Taq II</td><td>Takara Biomedical Technology (Beijing) Co., Ltd</td><td>RR820A</td><td></td></tr><tr><td>PrimeScript RT reagent kit with gDNA Eraser</td><td>Takara Biomedical Technology (Beijing) Co., Ltd</td><td>RR047A</td><td></td></tr><tr><td>Fully Enclosed Tissue Processor</td><td>Leica Biosystems Nussloch GmbH</td><td>ASP200S</td><td></td></tr><tr><td>Heated Paraffin Embedding Module</td><td>Leica Biosystems Nussloch GmbH</td><td>EG1150H</td><td></td></tr><tr><td>Semi-Automated Rotary Microtome</td><td>Leica Biosystems Nussloch GmbH</td><td>RM2245</td><td></td></tr><tr><td>Water bath for paraffin sections</td><td>Leica Biosystems Nussloch GmbH</td><td>HI1210</td><td></td></tr><tr><td>Autostainer XL</td><td>Leica Biosystems Nussloch GmbH</td><td>ST5010</td><td></td></tr><tr><td>Agilent 2100</td><td>Agilent Technologies</td><td>G2939A</td><td></td></tr><tr><td>Optical microscope</td><td>Olympus</td><td>BX51</td><td></td></tr></tbody></table>

intracranial subarachnoidal route of infection for investigating roles of streptococcus suis biofilms in meningitis in a mouse infection model

猪链球菌不仅是全球猪的主要细菌病原体, 而且是一种新兴的人畜共患病剂。在人类和猪中, 脑膜炎是感染猪的主要表现。适当的感染模型是了解病原体致病机制的基本工具。研究了小鼠感染的几种途径, 探讨了猪感染的发病机制。然而, 腹腔, 鼻腔和静脉感染的途径不适合研究猪的表面成分在脑膜炎直接在脑中的作用, 如生物膜胞外基质。虽然 intracisternal 接种已被用于猪的感染, 但精确注射部位尚未描述。本文用小鼠模型描述了颅内 subarachnoidal 感染的途径, 探讨了生物膜在猪链球菌脑膜炎中的作用。通过位于 bregma 3.5 毫米延髓的注射部位, 将猪的浮游细胞或生物膜状态细胞直接注入小鼠蛛网膜下腔。用生物膜状态细胞注射小鼠脑组织中 TLR2 和细胞因子的病理学分析和 mRNA 表达明显表明,猪的生物膜在猪的脑膜炎中起着决定性的作用。这种感染途径明显优于其他感染途径, 允许研究宿主-细菌相互作用。此外, 它允许细菌成分对宿主免疫应答的影响直接在大脑被评估, 并模仿细菌进入中枢神经系统。这种感染途径可以扩展, 以调查其他细菌引起的脑膜炎机制。此外, 还可用于检测药物对细菌性脑膜炎的疗效。

在实验条件下对生物膜的形成进行了 SEM 分析。如图 1所示, 浮游细胞 (图 1a) 与生物膜状态细胞之间的生物膜形成有显著差异 (图 1B)。SEM 分析表明, 生物膜细菌在团簇和多层, 它们被包裹在细胞外基质, 而浮游细菌的密度更低, 主要分散单独。
<p class="jove_content" fo:keep-together.within-...

Watch this Scientific Journal Video about Intracranial Subarachnoidal Route of Infection for Investigating Roles of Streptococcus suis Biofilms in Meningitis in a Mouse Infection Model at JoVE.com

Intracranial Subarachnoidal Route of Infection for Investigating Roles of Streptococcus suis Biofilms in Meningitis in a Mouse Infection Model

本文描述了小鼠颅内 subarachnoidal 感染的途径, 研究了生物膜在猪链球菌脑膜炎中的作用。这种感染模型也适用于研究其他细菌性脑膜炎的发病机制和新药对细菌性脑膜炎的疗效。

颅内 Subarachnoidal 感染对猪链球菌生物膜在小鼠感染模型中作用的研究

Streptococcus suis is not only a major bacterial pathogen of pigs worldwide but also an emerging zoonotic agent. In humans and pigs, meningitis is a major ...

intracranial-subarachnoidal-route-infection-for-investigating-roles

Universidad de Cartagena UDEC

Research

JoVE Journal

Immunology and Infection

Intracranial Subarachnoidal Route of Infection for Investigating Roles of Streptococcus suis Biofilms in Meningitis in a Mouse Infection Model

7.2K 次观看.  Nanjing Agricultural University. 本文描述了小鼠颅内 subarachnoidal 感染的途径, 研究了生物膜在猪链球菌脑膜炎中的作用。这种感染模型也适用于研究其他细菌性脑膜炎的发病机制和新药对细菌性脑膜炎的疗效。

视频: Intracranial Subarachnoidal Route of Infection for Investigating Roles of Streptococcus suis Biofilms in Meningitis in a Mouse Infection Model

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 ...

科研

免疫与感染

A Murine Model of Subarachnoid Hemorrhage

In this video publication a standardized mouse model of subarachnoid hemorrhage (SAH) is presented. Bleeding is induced by endovascular Circle of Willis perforation (CWp) and proven by intracranial pressure (ICP) monitoring. Thereby a homogenous blood distribution in subarachnoid spaces surrounding the arterial circulation and cerebellar fissures is achieved. Animal physiology is maintained by intubation, mechanical ventilation, and continuous on-line monitoring of various physiological and cardiovascular parameters: body temperature, systemic blood pressure, heart rate, and hemoglobin saturation. Thereby the cerebral perfusion pressure can be tightly monitored resulting in a less variable volume of extravasated blood. This allows a better standardization of endovascular filament perforation in mice and makes the whole model highly reproducible. Thus it is readily available for pharmacological and pathophysiological studies in wild type and genetically altered mice.

A standardized mouse model of subarachnoid hemorrhage by intraluminal Circle of Willis perforation is described. Vessel perforation and subarachnoid bleeding are monitored by intracranial pressure monitoring. In addition various vital parameters are recorded and controlled to maintain physiologic conditions.

蛛网膜下腔出血的小鼠模型

In this video publication a standardized mouse model of subarachnoid  hemorrhage (SAH) is presented. Bleeding is induced by endovascular Circle of  Willis ...

医学

Visualization of Streptococcus pneumoniae within Cardiac Microlesions and Subsequent Cardiac Remodeling

During bacteremia Streptococcus pneumoniae can translocate across the vascular endothelium into the myocardium and form discrete bacteria-filled microscopic lesions (microlesions) that are remarkable due to the absence of infiltrating immune cells. Due to their release of cardiotoxic products, S. pneumoniae within microlesions are thought to contribute to the heart failure that is frequently observed during fulminate invasive pneumococcal disease in adults. Herein is demonstrated a protocol for experimental mouse infection that leads to reproducible cardiac microlesion formation within 30 hr. Instruction is provided on microlesion identification in hematoxylin &#38; eosin stained heart sections and the morphological distinctions between early and late microlesions are highlighted. Instruction is provided on a protocol for verification of S. pneumoniae within microlesions using antibodies against pneumococcal capsular polysaccharide and immunofluorescent microscopy. Last, a protocol for antibiotic intervention that rescues infected mice and for the detection and assessment of scar formation in the hearts of convalescent mice is provided. Together, these protocols will facilitate the investigation of the molecular mechanisms underlying pneumococcal cardiac invasion, cardiomyocyte death, cardiac remodeling as a result of exposure to S. pneumoniae, and the immune response to the pneumococci in the heart.

Visualization of Streptococcus pneumoniae within Cardiac Microlesions and Subsequent Cardiac Remodeling

Streptococcus pneumoniae forms discrete non-purulent microscopic lesions in the heart. Outlined is the protocol for a murine model of cardiac microlesion formation. Instruction is provided on microlesion visualization using microscopy, discrimination between early and late microlesions, and methods to detect cardiac remodeling in hearts of convalescent animals.

可视化<em&gt;肺炎链球菌</em&gt;在心脏Microlesions和随后的心脏重构

During bacteremia Streptococcus pneumoniae can translocate across the vascular endothelium into the myocardium and form discrete bacteria-filled ...

A Mouse Model to Assess Innate Immune Response to Staphylococcus aureus Infection

Staphylococcus aureus (S. aureus)&#160;infections, including methicillin resistant stains, are an enormous burden on the healthcare system. With incidence rates of S. aureus infection climbing annually, there is a demand for additional research in its&#160;pathogenicity. Animal models of infectious disease advance our understanding of the host-pathogen response and lead to the development of effective therapeutics. Neutrophils play a primary role in the innate immune response that controls S. aureus infections by forming an abscess to wall off the infection and facilitate bacterial clearance; the number of neutrophils that infiltrate an S. aureus skin infection often correlates with disease outcome. LysM-EGFP mice, which possess the enhanced green fluorescent protein (EGFP) inserted in the Lysozyme M (LysM) promoter region (expressed primarily by neutrophils), when used in conjunction with in vivo whole animal fluorescence imaging (FLI) provide&#160;a means of quantifying neutrophil emigration noninvasively and longitudinally into wounded skin. When combined with a bioluminescent S. aureus strain and sequential in vivo whole animal bioluminescent imaging (BLI), it is possible to longitudinally monitor both the neutrophil recruitment dynamics and in vivo bacterial burden at the site of infection in anesthetized mice from onset of infection to resolution or death. Mice are more resistant to a number of virulence factors produced by S. aureus that facilitate effective colonization and infection in humans. Immunodeficient mice provide a more sensitive animal model to examine persistent S. aureus infections and the ability of therapeutics to boost innate immune responses. Herein, we characterize responses in LysM-EGFP mice that have been bred to MyD88-deficient mice (LysM-EGFP&#215;MyD88-/- mice) along with wild-type LysM-EGFP mice to investigate S. aureus skin wound infection. Multispectral simultaneous detection enabled study of neutrophil recruitment dynamics by using in vivo FLI, bacterial burden by using in vivo BLI, and wound healing longitudinally and noninvasively over time.

A Mouse Model to Assess Innate Immune Response to Staphylococcus aureus Infection

An approach is described for real-time detection of the innate immune response to cutaneous wounding and Staphylococcus aureus infection of mice. By comparing LysM-EGFP mice (which possess fluorescent neutrophils) with a LysM-EGFP crossbred immunodeficient mouse strain, we advance our understanding of infection and the development of approaches to combat infection.

一种评价金黄色葡萄球菌感染先天免疫反应的小鼠模型

Staphylococcus aureus (S. aureus)&#160;infections, including methicillin resistant stains, are an enormous burden on the healthcare system. With incidence ...

Inducing Meningococcal Meningitis Serogroup C in Mice via Intracisternal Delivery

Neisseria meningitidis (meningococcus) is a narrow-host-range microorganism, globally recognized as the leading cause of bacterial meningitis. Meningococcus is a transient colonizer of human nasopharynx of approximately 10% of healthy subject. In particular circumstances, it acquires an invasive ability to penetrate the mucosal barrier and invades the bloodstream causing septicaemia. In the latest case, fulminating sepsis could arise even without the consequent development of meningitis. Conversely, bacteria could poorly multiply in the bloodstream, cross the blood brain barrier, reach the central nervous system, leading to fulminant meningitis. The murine models of bacterial meningitis represent a useful tool to investigate the host-pathogen interactions and to analyze the pathogenetic mechanisms responsible for this lethal disease. Although, several experimental model systems have been evaluated over the last decades, none of these were able to reproduce the characteristic pathological events of meningococcal disease. In this experimental protocol, we describe a detailed procedure for the induction of meningococcal meningitis in a mouse model based on the intracisternal inoculation of bacteria. The peculiar signs of human meningitis were recorded in the murine host through the assessment of clinical parameters (e.g., temperature, body weight), evaluation of survival rate, microbiological analysis and histological examination of brain injury. When using intracisternal (i.cist.) inoculum, meningococci complete delivery directly into cisterna magna, leading to a very efficient meningococcal replication in the brain tissue. A 1,000-fold increase of viable count of bacteria is observed in about 18 h. Moreover, meningococci are also found in the spleen, and liver of infected mice, suggesting that the liver may represent a target organ for meningococcal replication.

Here, we describe a method to induce meningococcal meningitis through an intracisternal route of infection in adult mice. We present a step by step protocol of meningococcal infection from the preparation of inoculum to the intracisternal infection; then record the animal survival and evaluate the bacterial loads in murine tissues.

通过细胞内分娩诱导小鼠脑膜炎球菌性脑膜炎血清群C

Neisseria meningitidis (meningococcus) is a narrow-host-range microorganism, globally recognized as the leading cause of bacterial meningitis. ...

A Mouse Model for the Transition of Streptococcus pneumoniae from Colonizer to Pathogen upon Viral Co-Infection Recapitulates Age-Exacerbated Illness

Streptococcus pneumoniae (pneumococcus) is an asymptomatic colonizer of the nasopharynx in most individuals but can progress to a pulmonary and systemic pathogen upon influenza A virus (IAV) infection. Advanced age enhances host susceptibility to secondary pneumococcal pneumonia and is associated with worsened disease outcomes. The host factors driving those processes are not well defined, in part due to a lack of animal models that reproduce the transition from asymptomatic colonization to severe clinical disease. 
This paper describes a novel mouse model that recreates the transition of pneumococci from asymptomatic carriage to disease upon viral infection. In this model, mice are first intranasally inoculated with biofilm-grown pneumococci to establish asymptomatic carriage, followed by IAV infection of both the nasopharynx and lungs. This results in bacterial dissemination to the lungs, pulmonary inflammation, and obvious signs of illness that can progress to lethality. The degree of disease is dependent on the bacterial strain and host factors. 
Importantly, this model reproduces the susceptibility of aging, because compared to young mice, old mice display more severe clinical illness and succumb to disease more frequently. By separating carriage and disease into distinct steps and providing the opportunity to analyze the genetic variants of both the pathogen and the host, this S. pneumoniae/IAV co-infection model permits the detailed examination of the interactions of an important pathobiont with the host at different phases of disease progression. This model can also serve as an important tool for identifying potential therapeutic targets against secondary pneumococcal pneumonia in susceptible hosts.

A Mouse Model for the Transition of Streptococcus pneumoniae from Colonizer to Pathogen upon Viral Co-Infection Recapitulates Age-Exacerbated Illness

This paper describes a novel mouse model for the transition of pneumococcus from an asymptomatic colonizer to a disease-causing pathogen during viral infection. This model can be readily adapted to study polymicrobial and host-pathogen interactions during the different phases of disease progression and across various hosts.

病毒合并感染后 肺炎链球菌 从定植者转变为病原体的小鼠模型概括了年龄加重的疾病

Streptococcus pneumoniae (pneumococcus) is an asymptomatic colonizer of the nasopharynx in most individuals but can progress to a pulmonary and systemic ...

In Vivo Mouse Model of Spinal Implant Infection

Spine implant infections portend poor outcomes as diagnosis is challenging and surgical eradication is at odds with mechanical spinal stability. The purpose of this method is to describe a novel mouse model of spinal implant infection (SII) that was created to provide an inexpensive, rapid, and accurate in vivo tool to test potential therapeutics and treatment strategies for spinal implant infections.
In this method, we present a model of posterior-approach spinal surgery in which a stainless-steel k-wire is transfixed into the L4 spinous process of 12-week old C57BL/6J wild-type mice and inoculated with 1 x 103 CFU of a bioluminescent strain of Staphylococcus aureus Xen36 bacteria. Mice are then longitudinally imaged for bioluminescence in vivo on post-operative days 0, 1, 3, 5, 7, 10, 14, 18, 21, 25, 28, and 35. Bioluminescence imaging (BLI) signals from a standardized field of view are quantified to measure in vivo bacterial burden.
To quantify bacteria adhering to implants and peri-implant tissue, mice are euthanized and the implant and surrounding soft tissue are harvested. Bacteria are detached from the implant by sonication, cultured overnight and then colony forming units (CFUs) are counted. The results acquired from this method include longitudinal bacterial counts as measured by in vivo S. aureus bioluminescence (mean maximum flux) and CFU counts following euthanasia.
While prior animal models of instrumented spine infection have involved invasive, ex vivo tissue analysis, the mouse model of SII presented in this paper leverages noninvasive, real time in vivo optical imaging of bioluminescent bacteria to replace static tissue study. Applications of the model are broad and may include utilizing alternative bioluminescent bacterial strains, incorporating other types of genetically engineered mice to contemporaneously study host immune response, and evaluating current or investigating new diagnostic and therapeutic modalities such as antibiotics or implant coatings.

The protocol describes a novel in vivo mouse model of spinal implant infection where a stainless-steel k-wire implant is infected with bioluminescent Staphylococcus aureus Xen36. Bacterial burden is monitored longitudinally with bioluminescent imaging and confirmed with colony forming unit counts after euthanasia.

脊柱植入物感染的体内小鼠模型

Spine implant infections portend poor outcomes as diagnosis is challenging and surgical eradication is at odds with mechanical spinal stability. The ...

<meta charset="utf8"></head><body>用于细胞内 金黄色葡萄球菌 感染研究的优化体内模型

Development and Assessment of Intracellular Infection Models for Staphylococcus aureus

S. aureus can invade and persist within host cells, including immune cells, which allows it to evade immune detection and clearance. This intracellular persistence contributes to chronic and recurrent infections, complicating treatment and prolonging the disease. Consequently, there is a critical need for an intracellular infection model to better understand, prevent, and treat infections caused by S. aureus. This study indicated that antibiotics effectively eliminated extracellular bacteria but could not eradicate those that had entered the cells. Thus, a stable intracellular infection in vitro was established by RAW264.7 infected with S. aureus and co-culturing them with antibiotics. Subsequently, an intracellular infection model in mice was established by injecting peritoneal macrophages containing the intracellular infection. Vancomycin effectively cleared bacterial loads in mice challenged with planktonic S. aureus; however, it was ineffective against mice infected with equal or lower levels of intracellular bacteria within the peritoneal macrophages. This indicates that the intracellular infection model of S. aureus was successfully established, offering potential insights for the prevention and treatment of intracellular infections.

<meta charset="utf8"></head><body>金黄色葡萄球菌细胞内感染模型的开发与评价

Staphylococcus aureus (S. aureus) has the capability to disseminate throughout the body, causing persistent and recurrent infections. To better understand these processes, this study establishes an intracellular infection model for S. aureus. This model will provide a crucial foundation for investigating the mechanisms behind intracellular infections.

金黄色葡萄球菌细胞内感染模型的开发与评价

S. aureus can invade and persist within host cells, including immune cells, which allows it to evade immune detection and clearance. This intracellular ...

颅内 Subarachnoidal 感染对猪链球菌生物膜在小鼠感染模型中作用的研究

摘要

探索更多视频

此视频中的章节

人源化的小鼠模型来研究细菌感染瞄准微血管

蛛网膜下腔出血的小鼠模型

可视化

一种评价金黄色葡萄球菌感染先天免疫反应的小鼠模型

通过细胞内分娩诱导小鼠脑膜炎球菌性脑膜炎血清群C

病毒合并感染后肺炎链球菌从定植者转变为病原体的小鼠模型概括了年龄加重的疾病

脊柱植入物感染的体内小鼠模型

金黄色葡萄球菌细胞内感染模型的开发与评价

金黄色葡萄球菌细胞内感染模型的开发与评价

金黄色葡萄球菌细胞内感染模型的开发与评价

颅内 Subarachnoidal 感染对猪链球菌生物膜在小鼠感染模型中作用的研究

摘要

探索更多视频

此视频中的章节

人源化的小鼠模型来研究细菌感染瞄准微血管

蛛网膜下腔出血的小鼠模型

可视化

一种评价金黄色葡萄球菌感染先天免疫反应的小鼠模型

通过细胞内分娩诱导小鼠脑膜炎球菌性脑膜炎血清群C

病毒合并感染后 肺炎链球菌 从定植者转变为病原体的小鼠模型概括了年龄加重的疾病

脊柱植入物感染的体内小鼠模型

金黄色葡萄球菌细胞内感染模型的开发与评价

金黄色葡萄球菌细胞内感染模型的开发与评价

金黄色葡萄球菌细胞内感染模型的开发与评价

病毒合并感染后肺炎链球菌从定植者转变为病原体的小鼠模型概括了年龄加重的疾病