Name: passive administration of monoclonal antibodies against h. capsulatum and others fungal pathogens
Uploaded: 2011-02-14T17:00:00
Description: Watch this Scientific Journal Video about Passive Administration of Monoclonal Antibodies Against H. capsulatum and Others Fungal Pathogens at JoVE.com

1。生长的H. Capsulatum <ol><li>与真菌的工作准备了一个生物安全内阁，内阁由20分钟的紫外线照射10％的漂白粉清洗。H.在酵母阶段的 capsulatum 需要生物安全水平（BSL）二做法，而模具的形式要求BSLIII。使用酵母的形式，在下面的步骤。 </li><li>下一步准备用5毫升的PBS 15 ml锥形管。收获了殖民地的H. capsulatum（株HC ATCC G217B）从一个BHI血琼脂平板[葡萄糖10 g...

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	<li>Casadevall, A., Mukherjee, J., Scharff, M. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Monoclonal+antibody+based+ELISAs+for+cryptococcal+polysaccharide.">Monoclonal antibody based ELISAs for cryptococcal polysaccharide.</a> J. Immunol. Methods. 154, 27-35 (1992).</li><li>Guimaraes, A. J., Frases, S., Gomez, F. J., Zancope-Oliveira, R. M., Nosanchuk, J. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Monoclonal+antibodies+to+heat+shock+protein+60+alter+the+pathogenesis+of+Histoplasma+capsulatum.+Infect+Immun+77.">Monoclonal antibodies to heat shock protein 60 alter the pathogenesis of Histoplasma capsulatum. Infect Immun 77.</a> , 1357-1367 (2009).</li><li>Guimaraes, A. J., Hamilton, A. J., de M Guedes, H. L., Nosanchuk, J. D., Zancope-Oliveira, R. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Biological+function+and+molecular+mapping+of+M+antigen+in+yeast+phase+of+Histoplasma+capsulatum.">Biological function and molecular mapping of M antigen in yeast phase of Histoplasma capsulatum.</a> PLoS One. 3, e3449-e3449 (2008).</li><li>Nosanchuk, J. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Protective+antibodies+and+endemic+dimorphic+fungi.">Protective antibodies and endemic dimorphic fungi.</a> Curr Mol Med. 5, 435-442 (2005).</li><li>Nosanchuk, J. D., Steenbergen, J. N., Shi, L., Jr, D. e. e. p. e., &amp;amp, G. S., Casadevall, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Antibodies+to+a+cell+surface+histone-like+protein+protect+against+Histoplasma+capsulatum.">Antibodies to a cell surface histone-like protein protect against Histoplasma capsulatum.</a> J Clin Invest. 112, 1164-1175 (2003).</li><li>Nosanchuk, J. D., Steenbergen, J. N., Shi, L., Deepe, G. S., , ., Casadevall, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Antibodies+to+a+cell+surface+histone-like+protein+protect+against+Histoplasma+capsulatum.">Antibodies to a cell surface histone-like protein protect against Histoplasma capsulatum.</a> J. Clin. Invest. 112, 1164-1175 (2003).</li><li>Smith, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Responses+of+laboratory+animals+to+some+injectable+anaesthetics.">Responses of laboratory animals to some injectable anaesthetics.</a> Lab Anim. 27, 30-39 (1993).</li></ol>

这里介绍的协议表明，单克隆抗体的到 H capsulatum可以修改实验小鼠组织胞浆菌病的过程中。病原菌的细胞表面抗原的单克隆抗体可以修改主机和病原体之间的相互作用过程中发生的复杂的动态。这项研究建立了单克隆抗体介导腹腔注射时的一个致命性组织胞浆菌病的小鼠模型的保护，并建议候选蛋白疫苗的开发，如H2B，中号抗原（表面过氧化氢酶）（3）和HSP60。管理HC HSP60和H2B保护?...

<table border="1"><tbody><tr><th>材料名称</th><th>公司</th></tr><tr><td>生物安全柜（BSL2） </td><td></td></tr><tr><td> 15 mL锥形管</td><td>猎鹰，BD </td></tr><tr><td>火腿的F - 12培养基</td><td> Gibco公司</td></tr><tr><td> 37 °彗星摇床</td><td></td></tr><tr><td>涡旋</td><td></td></tr><tr><td> 50 ml锥形管</td><td>猎鹰，BD </td></tr><tr><td> 26G1 / 2针</td><td>屋宇署</td></tr><tr><td> 10 mL注射器</td><td>屋宇署</td></tr><tr><td> 250毫升烧瓶</td><td></td></tr><tr><td> FPLC（ 快速蛋白液相色谱法）系统</td><td>通用电气Helthcare </td></tr><tr><td>酶标仪</td><td>百特</td></tr><tr><td>麻醉（氯胺酮和甲苯噻嗪） </td><td></td></tr><tr><td>列代表</td><td></td></tr><tr><td>尼龙绳</td><td></td></tr><tr><td>散热灯</td><td></td></tr><tr><td> 70μm细胞过滤器</td><td>猎鹰，BD </td></tr><tr><td> BHI琼脂平板</td><td> Gibco公司</td></tr></tbody></table>

passive administration of monoclonal antibodies against h. capsulatum and others fungal pathogens

使用这种方法的目的是1），以推进我们的能力，以保护个人与抗体或疫苗，以防止或治疗由真菌组织胞浆菌和2）审查致病因素的作用，为治疗目标引起的组织胞浆菌病。产生单克隆抗体，免疫小鼠的免疫反应是采用固相酶联免疫吸附在我们的实验室开发的系统进行评估，并选择隔离的脾细胞与杂交瘤细胞融合的最好的响应者小鼠。 C57BL / 6小鼠已被广泛用于研究H。 capsulatum的发病机制，并提供获得所需的数据的最佳模式。为了评估在感染的单克隆抗体的作用，小鼠腹腔要么单抗管理到 H capsulatum或亚型匹配控制单克隆抗体，然后感染要么静脉（IV），腹腔（IP），或滴鼻（中）路线。在科学文献中，单克隆抗体在小鼠体内的真菌感染疗效依赖对死亡率作为终点，在殖民地成蛋白单位（CFU）在较早的时间点评估一起。生存（死亡时间）的研究是必要的，因为他们最能代表人类疾病。因此，我们干预的疗效不会充分地建立无生存曲线。这也是真正建立疫苗疗效或毒力突变体的测试。随着组织胞浆菌病，老鼠经常去充满活力，以死了几个小时。最初可能会干预，如管理一个MAB能力保护的动物疾病，但疾病的复发这不会在短期CFU实验实现。除了生存和真菌的负担分析，我们考察感染（蜂窝招聘，组织学，细胞因子反应）的炎症反应。生存/死亡实验，小鼠被感染，至少每天两次监测发病的迹象。为了评估真菌的负担，组织病理学和细胞因子的反应，小鼠在感染后的不同时期，实施安乐死。根据阿尔伯特爱因斯坦医学院的动物实验研究所的指引，进行动物实验。

Watch this Scientific Journal Video about Passive Administration of Monoclonal Antibodies Against H. capsulatum and Others Fungal Pathogens at JoVE.com

Passive Administration of Monoclonal Antibodies Against H. capsulatum and Others Fungal Pathogens

C57BL / 6小鼠已被用于研究HC发病机制，并提供最好的模式。我们正在探索这种真菌对体液免疫的潜在利益，并产生了几个单克隆抗体[组蛋白H2B和热休克蛋白60kDa]，我们为他们的腹腔给药后的保护效果测试。

单克隆抗体的反对消极行政 H。 capsulatum和其他真菌病原体

The purpose of the use of this methodology is 1) to advance our capacity to protect individuals with antibody or vaccine for preventing or treating ...

Research

JoVE Journal

Immunology and Infection

Passive Administration of Monoclonal Antibodies Against H. capsulatum and Others Fungal Pathogens

Quantitative Analyses of all Influenza Type A Viral Hemagglutinins and Neuraminidases using Universal Antibodies in Simple Slot Blot Assays

所有流感的定量分析类型使用通用抗体的一种病毒Hemagglutinins和Neuraminidases，在简单的狭缝杂交分析

Hemagglutinin (HA) and neuraminidase (NA) are two surface proteins of influenza viruses which are known to play important roles in the viral life cycle ...

科研

免疫与感染

Infection of Zebrafish Embryos with Intracellular Bacterial Pathogens

斑马鱼的胚胎细胞内细菌病原体感染

Zebrafish (Danio rerio) embryos are increasingly used as a model for studying the function of the vertebrate innate immune system in host-pathogen ...

Assessing Anti-fungal Activity of Isolated Alveolar Macrophages by Confocal Microscopy

评估隔离肺泡巨噬细胞的激光共聚焦显微镜抗真菌活性

The lung is an interface where host cells are routinely exposed to microbes and microbial products. Alveolar macrophages are the first-line phagocytic ...

Biolistic Transformation of a Fluorescent Tagged Gene into the Opportunistic Fungal Pathogen Cryptococcus neoformans

Biolistic Transformation of a Fluorescent Tagged Gene into the Opportunistic Fungal Pathogen Cryptococcus neoformans

荧光标记基因的基因枪转化成机会性真菌病原<em&gt;新型隐球菌</em

The basidiomycete Cryptococcus neoformans, an invasive opportunistic pathogen of the central nervous system, is the most frequent cause of fungal ...

Generation of Recombinant Human IgG Monoclonal Antibodies from Immortalized Sorted B Cells

从排序的永生B细胞产生的重组人IgG单克隆抗体

Finding new methods for generating human monoclonal antibodies is an active research field that is important for both basic and applied sciences, ...

Generation of Murine Monoclonal Antibodies by Hybridoma Technology

鼠单克隆抗体的产生杂交瘤技术

Monoclonal antibodies are universal binding molecules and are widely used in biomedicine and research. Nevertheless, the generation of these binding ...

A Robust Pneumonia Model in Immunocompetent Rodents to Evaluate Antibacterial Efficacy against S. pneumoniae, H. influenzae, K. pneumoniae, P. aeruginosa or A. baumannii

A Robust Pneumonia Model in Immunocompetent Rodents to Evaluate Antibacterial Efficacy against S. pneumoniae, H. influenzae, K. pneumoniae, P. aeruginosa or  A. baumannii

在免疫鼠害的鲁棒性肺炎模型评价抗菌功效反对<i&gt;秒。肺炎</i&gt;<i&gt;小时。流感</i&gt;<i&gt; K。肺炎</i&gt;<em&gt;点。铜绿假单胞菌</em&gt;或<em&gt;一个。鲍曼不动杆菌</em

Efficacy of candidate antibacterial treatments must be demonstrated in animal models of infection as part of the discovery and development process, ...

Generation of Escape Variants of Neutralizing Influenza Virus Monoclonal Antibodies

中和流感病毒单克隆抗体逃逸变种的产生

Influenza viruses exhibit a remarkable ability to adapt and evade the host immune response. One way is through antigenic changes that occur on the surface ...

Generation of Discriminative Human Monoclonal Antibodies from Rare Antigen-specific B Cells Circulating in Blood

Monoclonal antibodies (mAbs) are powerful tools useful for both fundamental research and in biomedicine. Their high specificity is indispensable when the ...

Generation of Monoclonal Antibodies Against Natural Products

针对天然产物的单克隆抗体的产生

The analysis of the bioactive components present in foods and natural products has become a popular area of study in many fields, including traditional ...