我们感谢的托德Schlenke教授，教授托尼IP转基因果蝇株，为Trichopria果蝇和教授卡尔桥本反Spätzle抗体。此演示实验室的贡献，我们感谢过去和现在的成员。支持这项工作是由下列补助：（S06 GM08168，高层41399-009，和G12-RR03060），，从国立卫生研究院，美国农业部（野村综合研究所/农业部CSREES 2006-03817和2009-35302-05277）和PSC-纽约市立大学。

整个实验的协议分为四个步骤（ 图9）。 （1）蝇蛆养殖黄蜂（2）建立感染和准备解剖动物;（3）隔离和修复主机/寄生虫结构;（4）免疫组织分析。 1。对果蝇幼虫培养黄蜂维护黄蜂文化需要仔细规划。相对于日益增长的苍蝇，这是相当劳力密集。我们维持我们在实验室YW应变果蝇幼虫或蛹的蜂殖民地在24°C培养黄蜂保持在合适的阶段，连续源的主机，并清除“感染”瓶苍蝇出现之前，黄蜂做。黄蜂按照性别决定haplodiploid方法，因此，它是重要的雌性交配之前，他们感染的主机。 <ol><li>第1天，添加少量的新鲜前民建联相比酵母膏（1毫升的水混合在大约1活性干酵母到一个新飞食品小瓶Ğ飞食品的配方是不是关键。我们成长上corn-meal/sucrose/agar介质的苍蝇。地方50-60年轻（2-4日龄）成人YW（或其他本质上是野生型株）飞行打下48小时在24°C蛋</li><li>第3天，取出小瓶苍蝇。与蜂蜜下降的嗡嗡声插头内侧的小瓶放在一个时髦的插件。蜂蜜中的蒸馏水稀释2:1。 </li><li>使用的CO 2，麻醉黄蜂和排序6-8名女性和6-8男性，大约2-3天。将它们添加到小瓶含0-48小时的旧主机。请注意，黄蜂更敏感，对CO 2比苍蝇，你可能要校准他们接触到的气体。 果蝇的研究人员不使用标准的果蝇麻醉站设计。一个很好的起点，麻醉黄蜂设置的CO 2压力大约一半需要麻醉测定苍蝇。 </li><li>更换小瓶的嗡嗡声插头（用蜂蜜）。轻轻放置在其一侧的小瓶直到黄蜂醒来。 </li><li>蝇的幼虫和蜂在29°C培养箱放置这些小瓶。 </li><li>成功的感染会略有延迟pupariation。主机遭受马蜂攻击，但都能够为自己辩护（或如果他们不感染），继续发展后，正常的作息时间和成蝇从这些puparia出现以及未来的黄蜂。 </li><li>根据蜂和温度，黄蜂eclose在25-30天。 </li><li>从小瓶蜂在发展成蝇。 </li></ol> 2。设立感染和准备受感染的动物解剖在开始之前，有几个干净的玻璃或塑料培养皿中，用Pyrex解剖盘9洼地，Kimwipes，蒸馏水（在喷瓶），70％的乙醇（在喷瓶），1X PBS（在喷瓶），显微镜幻灯片，小，清洁刮刀方便。 <ol><li>设立感染，按照上述步骤1-5。根据实验，它可能需要使用主机在大致相同的发展阶段。对于这一点，egglays 2-6小时，可用于感染。 </li><li>收获感染的幼虫（解剖免疫组织或寄生虫），成一个小的玻璃或塑料培养皿中删除蝇小瓶的内容。 </li><li>使用体视显微镜和钳子（镊子）的罚款，精心挑选6-10幼虫，一个由一，他们先用1X PBS将陷入萧条，然后转移到水，70％的乙醇，水和1×PBS，先后。这些步骤的目的是摆脱苍蝇食物的动物，并彻底清洗和消毒其表面。冲洗水稀释乙醇和最终冲洗PBS中删除的乙醇。下面的步骤3，最好不要离开在PBS动物的时间超过10分钟。 </li></ol> 3。隔离并修复主机/寄生虫结构背景幼虫淋巴腺是一个小的造血器官7。在第三幼虫龄，淋巴腺包含了前叶的一双大背船只的侧翼（ 图3A，6B，7A - D）。具有独特的细胞特性的专门区域进一步分为前叶。三种类型的细胞，浆细胞，lamellocytes，晶体细胞的祖细胞位于前叶。心包细胞分离后从较小的叶前叶。 果蝇的淋巴腺是昆虫和哺乳动物的造血8,9模型。 脂肪体是哺乳动物的肝功能类似。体液免疫反应，引发肥胖的身体下面的微生物或黄蜂感染1,4,10。如因此，独特的抗菌肽基因组合被激活肽分泌到血淋巴1。肥胖的身体，也为基层组织糖原和甘油三酯的生产和储存11。幼虫的脂肪体占用大量的hemocoel量，所以不像淋巴腺，很容易找到。现在，我们将演示如何解剖的淋巴腺和三龄幼虫的脂肪体。 在您开始之前需要精细镊子（镊子）和乙醇清洗的载玻片。 幼虫淋巴腺清扫<ol><li>用细镊子，选择一个流浪的第三龄幼虫在1X PBS保存。 </li><li>放置到显微镜玻片上的幼虫。 </li><li>使用镊子，定位与腹侧方最多的动物。 </li><li>使用两对镊子，动物举行任一侧的后端（箭头1， 图3A）和gently拉角质层引入一个小的，在角质层表面的撕裂。 </li><li>血淋巴中含有血细胞，肠，脂肪体将开始逃离体腔。 10微升的“pipetteman”做涂片检查，如果需要的话可以采取的淋巴。 </li><li>口钩（箭头2， 图3A）动物右侧下配售两钳，轻轻地撕角质层。 </li><li>将两钳嘴钩（箭头2， 图3A）和动物的左侧下方，仔细再作小的撕裂。 </li><li>按住口动物钩，轻轻一拉角质层，对动物的后端（虽然脱皮）。 </li><li>由于角质层被拉回，脑，脂肪体，成虫盘，唾液腺，腺胃将成为暴露出来。分离角质层是在后端，但仍会有背船只与它相连的淋巴腺。&lt;/ LI&gt; <li>切开腺胃和移动了从淋巴腺区的。淋巴腺将下方的大脑，即使你可能无法看到它。 </li><li>仔细逗掉体内的脂肪，肠道，唾液腺，其他结构和腺体，可坐。要小心，不能脱离环腺和位于大脑后，背船只淋巴腺。淋巴腺将定位平对显微镜幻灯片。 </li><li>小心取出所有的其他组织的淋巴腺，直到整个腺前叶展开心包细胞决胜盘。 </li></ol> 注：正确解剖的淋巴腺将有一对前叶和两套沿背船只后叶（ 图6B）。叶可以很容易被损坏，或从背船只可以下降。样本，因此，应非常小心处理。腺也Ĥ作为枯萎或合同的倾向。轻轻地理顺机关在其后端允许所有部件和细胞以及提交​​。这是特别的免疫协议的必要。 仙女人体解剖<ol><li>蔡司1000解剖显微镜（解剖显微镜可用于任何立体声）解剖的阶段上，放置一个灯箱，使入射光通过样品传输。从第2步的三龄幼虫在200μL1×PBS显微镜幻灯片的位置。从样品的光源倾斜，使机体出现透明的，因此很容易可视其内部器官。 </li><li>使用镊子，位置，使前到底是离你的动物。 </li><li>使用镊子，保持角质层的幼虫其口钩左侧（箭头1， 图3B）。使用其他的镊子，轻轻地撕角质层后端的方式（ARrowhead 2， 图3B）。撕不从身体的角质层在后端。 </li><li>非常轻柔地开始消除肠道一侧。 </li><li>轻轻地取出大脑，成虫盘，环腺背船只运行与淋巴腺。 </li><li>不要取出唾液腺，是因为肥胖的身躯，坚持这些腺体。 </li><li>轻轻地去除角质层和肥胖的身体上留下的幻灯片在1X PBS。 </li></ol> 注1：脂肪体细胞层只有一个，因此，它是非常重要的所有细胞都在同一平面上的玻片夷为平地。细胞是endopolyploid的和容易想象。一个良好的脂肪体解剖样本应保持正常的细胞接触，应该有最小的脂肪球周围的解剖样本（ 图6I的J）。 注意2：如果蜂卵保持宿主的免疫系统的影响，他们将initiat电子商务的发展，几乎立即。寄生虫（主机幼虫）的早期发育阶段，很容易从的解剖hemocoel（ 图8）。寄生虫卵或幼虫坚持以脂肪体或其他器官，或简单的玻片上，在夹层滑动。 4。免疫组织分析<ol><li>空气干燥5-10分钟，从5-10分钟前准备4％多聚甲醛的PBS固定的脂肪体幻灯片删除1X PBS淋巴腺。每张幻灯片都可以有几个解剖染色淋巴腺。移动与固定样本后续步骤加湿室自制幻灯片。这个会议厅准备把两片折叠的湿纸巾，在一个空塑料微量提示框的底部。举行的提示使用分压器的顶部可放置一个或多个幻灯片样本。 </li><li>正因为如此，按照标准方法进行免疫组织分析结合在体内的细胞标记和间接免疫组化4。 </li></ol> 5。代表结果<ol><li>黄蜂感染激活Toll途径记者Drosomycin-GFP（ 图6G的J）的表达。在这个实验中基因表达的影响，黄蜂为患是在体内使用的GFP记者联系到DRS启动12可视化。在未受感染的控制动物，没有检测到GFP的表达。 GFP信号清楚地发现，从研究与感染的动物脂肪体细胞的细胞核，细胞质和解剖victoriae。为了取得最好的结果，黄蜂的比例：主机应该是1:10。感染应持续至少2个小时。后为患，DRS-GFP信号的加剧和许多脂肪体细胞的GFP阳性，甚至长达72个小时1,4。 </li><li>黄蜂感染诱导分化的lamellocytes在Lymph腺（ 图6D）。 lamellocytes包围和封锁黄蜂发展（ 图6H）。淋巴腺解剖可视化研究后前叶诱导细胞的变化victoriae感染。分化新lamellocytes是在解剖肺叶周边的; lamellocytes是由畸形的增强（MSNF9 GFP）驱动的GFP基因标记。使用罗丹明标记的鬼笔环肽与丝状肌动蛋白细胞可视化。请注意，前叶的完整性破坏，在这些腺体基底膜通常围绕着叶不断（ 图6C），现在是间断（ 图6D）。从相同的解剖，血淋巴lamellocytes可以也涂片中的可视化（ 图6G）;其中一些相关联的成立，以阻止寄生虫的发展（ 图6H）胶囊结构。 </li><li>小号pätzle蛋白表达在幼虫淋巴腺（ 图7C，D）的大多数细胞。检测解剖淋巴腺SPZ，我们遵循一个标准协议，染色的淋巴腺细胞与抗-SPZ抗体4。 在体内 ，SPZ水平提高后，黄蜂感染淋巴腺细胞（比较板图。7C，电子与7G，我和面板7D，7H F，歼 ）。 </li><li>黄蜂发展的早期和晚期阶段从寄主幼虫和蛹（ 图8）。检查蜂的发育阶段，受感染的幼虫在不同时间点解剖后产卵。在这里，我们显示了这些阶段Leptopilina菌采样。从被感染的野生型D。果蝇主持。 </li></ol><img alt="图1" src="/files/ftp_upload/3347/3347fig1.jpg" /> 图1。不同蜂种和蜂卵产卵。所有图片使用徕卡立体显微镜获得一个 果蝇Trichopria女性的黄蜂oviposits到了 D蛋果蝇蛹，B Trichopria果蝇产卵放大到主机蛹，C变暗点，表明在产卵部位的伤口愈合。 赉 。 boulardi男，E G. xanthopoda女性，F L. Ğ 研究victoriae女性。 heterotoma男（左）和女性。 <img alt="图2" src="/files/ftp_upload/3347/3347fig2.jpg" /> 图2。生命周期飞/黄蜂军备竞赛。产卵结果在两个成果之一。无论是宿主的免疫反应成功阻止发展黄蜂（左），或黄蜂阻挠宿主的免疫发源于NSES和成功（右）。从梅尔克和Govind，1999年第18改性。 <img alt="图3" src="/files/ftp_upload/3347/3347fig3.jpg" /> 图3。三龄幼虫显示感兴趣的器官前清扫。所有图片使用徕卡立体显微镜获得一个 HML&gt;绿色荧光蛋白与GFP（绿色）的幼虫淋巴腺单个细胞的荧光（箭头）表示一般的淋巴腺位置在一个完整的动物。荧光和亮场光学成像与幼虫。箭头指向乙 CG GFP绿色荧光蛋白表达在脂肪体表示的器官在一个完整的阿尼姆的位置幼虫为淋巴腺清扫描述的协议文本中的重要位置。等。箭头指向为脂肪体解剖描述的协议文本中的重要位置。 <img alt="图4" src="/files/ftp_upload/3347/3347fig4.jpg" /> 图4。免疫遗传主机电路对寄生蜂的攻击。Toll途径的核心组件的免疫反应 。被激活的蛋白酶Spätzle加工酶，SPEspätzle（SPZ）。 SPZ活性炭作为配收费。细胞内的信号，导致活化的NF-κB转录因子，背和DIF。仙人掌服务的途径IκB的抑制剂。 Drosomycin，规范收费通路靶基因激活，可以监控使用GFP报告基因的苍蝇株（见图6I的J）。 <img alt="图5" src="/files/ftp_upload/3347/3347fig5.jpg" /> 图5。在响应封装到 L victoriae为患需要蛇&gt; GFP-DL的果蝇主机。 蛇基因增强表达在血细胞13。当这个增强驱动背GFP的融合蛋白在转基因的表达，背一些浆细胞和lamellocytes弥补胶囊和聚合通过GFP的绿色荧光检测。所有图片使用蔡司​​LSM的共聚焦显微镜。 研究蛋样品victoriae。 屋宇署 Lamellocytes的（白色箭头）在后端的鸡蛋表达GFP的背。 光盘放大倍率越高，在面板B. EH样品counterstained罗丹明标记的鬼笔形象化丝状肌动蛋白（F-肌动蛋白，红）用Hoechst 33258可视化的DNA（蓝色），E L.封装鸡蛋victoriae 楼变暗胶囊的研究 生长激素 研究victoriae。 victoriae infection诱导血细胞聚集（plasmatocyte和lamellocyte）。 <img alt="图6" src="/files/ftp_upload/3347/3347fig6.jpg" /> 图6。免疫反应，反对黄蜂感染。面板中的图像，获得A和B使用蔡司​​Axio上的范围。 CJ-获得使用蔡司 ​​共聚焦显微镜LSM的板中的图像。寄主幼虫通过透明角质层显示变暗封装蜂卵（箭头）。 乙代表第三龄幼虫淋巴腺的Hoechst 33258染色显示穿插所有的叶和心包细胞的细胞。比例尺代表100μ。CJ-counterstained所有样品均用Hoechst 33258（标签原子核）和罗丹明鬼笔环肽（F-actin的标记）。CD前淋巴结腺叶对照组（C）和L. victoriae感染（四）MSNF9-GFP的动物。在受感染的动物，MSNF增强的表现，具体到洛杉矶mellocytes 14，核GFP记者发现，从感染的动物隔离é浆细胞不表达绿色荧光蛋白。这些动物也有极少数，如果的任何lamellocytes。F浆细胞（GFP阴性，短箭头） 属弱核GFP表达与分化新的，更大的lamellocytes（长箭头） victoriae感染的MSNF9-GFP的幼虫，G和聚合浆细胞和lamellocytes从一个研究 hemocoel感染victoriae-DRS-GFP的幼虫。 DRS-GFP记者表达被激活，在一些浆细胞（短箭头），但不是在lamellocytes（长箭头）感染后，H封装和研究变暗黄蜂幼虫boulardi感染MSNF9的GFP主机幼虫的。许多MSNF9 GFP阳性lamelloctyes环绕的黄蜂幼虫（黑暗的结构;粗箭头），并表现出强大的核GFP表达（长箭头）。这锅EL以前发表在PLoS ONE 15。IJ脂肪体细胞从感染的（我）和L.解剖victoriae感染（十）DRS-GFP幼虫。 GFP是在受感染的动物脂肪体细胞的核和细胞质。 <a href="http://www.jove.com/files/ftp_upload/3347/3347fig6_lg.jpg" target="_blank">点击这里查看大图</a> 。 <img alt="图7" src="/files/ftp_upload/3347/3347fig7.jpg" /> 图7。寄生蜂的感染。spätzle表达后欧塞尔样品counterstained的Hoechst 33258 DNA的可视化。所有使用蔡司​​LSM的共聚焦显微镜图像获得。从解剖未受感染的YW幼虫的自动对焦前叶。样品进行了处理，没有主要抗体的间接免疫染色（A和B）或抗的SPZ抗体（红色，C，D，E，F）和counterstained网站面粉鬼笔（绿色标签的F-肌动蛋白在细胞;B，D F）的吉焦前叶解剖感染YW幼虫和反SPZ抗体染色（红色，G，H，我十）的Alexa Fluor-鬼笔环肽（绿色;板Ĥ，歼） 外汇放大倍率越高，在C和D，分别IJ G和H样品的放大倍率较高，分别为样本。比例尺面板地下AD，H代表50微米。 <img alt="图8" src="/files/ftp_upload/3347/3347fig8.jpg" /> 图8。 L.幼虫和蛹的阶段heterotoma和L。分离幼虫或蛹飞的主机后为患boulardi。各个阶段，如所示。所有图片使用蔡司​​Axio上的范围。 连续自动对焦 L. heterotoma阶段，在感染后1-6天。 下方有一排自动对焦胚后L。 boulardi阶段，感染后4至12天。 <img alt="图9" sRC =“/ files/ftp_upload/3347/3347fig9.jpg”/&gt; 图9。实验协议的流程图。

<ol>
	<li>Schlenke, T. A., Morales, J., Govind, S., Clark, A. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Contrasting+infection+strategies+in+generalist+and+specialist+wasp+parasitoids+of+Drosophila+melanogaster.">Contrasting infection strategies in generalist and specialist wasp parasitoids of Drosophila melanogaster.</a> PLoS Pathog. 3, 1486-1501 (2007).</li><li>Chiu, H., Morales, J., Govind, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Identification+and+immuno-electron+microscopy+localization+of+p40,+a+protein+component+of+immunosuppressive+virus-like+particles+from+Leptopilina+heterotoma,+a+virulent+parasitoid+wasp+of+Drosophila.">Identification and immuno-electron microscopy localization of p40, a protein component of immunosuppressive virus-like particles from Leptopilina heterotoma, a virulent parasitoid wasp of Drosophila.</a> J. Gen. 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K., Hartenstein, V., Banerjee, U. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+hematopoietic+stem+cell+and+its+niche:+a+comparative+view.">The hematopoietic stem cell and its niche: a comparative view.</a> Genes Dev. 21, 3044-3060 (2007).</li><li>Lemaitre, B., Hoffmann, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+host+defense+of+Drosophila+melanogaster.">The host defense of Drosophila melanogaster.</a> Annu. Rev. Immunol. 25, 697-743 (2007).</li><li>Schlegel, A., Stainier, D. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Lessons+from+"lower"+organisms:+what+worms,+flies,+and+zebrafish+can+teach+us+about+human+energy+metabolism.">Lessons from "lower" organisms: what worms, flies, and zebrafish can teach us about human energy metabolism.</a> PLoS Genet. 3, e199 (2007).</li><li>Ferrandon, D., Jung, A. C., Criqui, M., Lemaitre, B., Uttenweiler-Joseph, S., Michaut, L., Reichhart, J., Hoffmann, J. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+drosomycin-GFP+reporter+transgene+reveals+a+local+immune+response+in+Drosophila+that+is+not+dependent+on+the+Toll+pathway.">A drosomycin-GFP reporter transgene reveals a local immune response in Drosophila that is not dependent on the Toll pathway.</a> EMBO J. 17, 1217-1227 (1998).</li><li>Bruckner, K., Kockel, L., Duchek, P., Luque, C. M., Rorth, P., Perrimon, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+PDGF/VEGF+receptor+controls+blood+cell+survival+in+Drosophila.">The PDGF/VEGF receptor controls blood cell survival in Drosophila.</a> Dev. Cell. 7, 73-84 (2004).</li><li>Tokusumi, T., Shoue, D. A., Tokusumi, Y., Stoller, J. R., Schulz, R. 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A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Characterization+of+a+lamellocyte+transcriptional+enhancer+located+within+the+misshapen+gene+of+Drosophila+melanogaster.">Characterization of a lamellocyte transcriptional enhancer located within the misshapen gene of Drosophila melanogaster.</a> PLoS One. 4, e6429 (2009).</li><li>Morales, J., Chiu, H., Oo, T., Plaza, R., Hoskins, S., Govind, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Biogenesis,+structure,+and+immune-suppressive+effects+of+virus-like+particles+of+a+Drosophila+parasitoid,+Leptopilina+victoriae.">Biogenesis, structure, and immune-suppressive effects of virus-like particles of a Drosophila parasitoid, Leptopilina victoriae.</a> J. Insect Physiol. 51, 181-195 (2005).</li><li>Bettencourt, R., Asha, H., Dearolf, C., Ip, Y. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hemolymph-dependent+and+-independent+responses+in+Drosophila+immune+tissue.">Hemolymph-dependent and -independent responses in Drosophila immune tissue.</a> J. Cell Biochem. 92, 849-863 (2004).</li><li>Melk, J. P., Govind, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Developmental+analysis+of+Ganaspis+xanthopoda,+a+larval+parasitoid+of+Drosophila+melanogaster.">Developmental analysis of Ganaspis xanthopoda, a larval parasitoid of Drosophila melanogaster.</a> J. Exp. Biol. 202, 1885-1896 (1999).</li></ol>

没有利益冲突的声明。

在对果蝇的寄生蜂的兴趣正在涌动解码整个基因组的分子生物学技术成为高效率和成本效益。然而，相对于他们非常出色的研究主机，蜂生物学的许多有趣的方面仍模糊不清。这些措施包括申办范围，免疫抑制，过寄生和行为有关的问题。这次演讲的重点是要证明在果蝇的免疫组织感染的影响。这里展示的解剖技术可以用于基因表达分析，RNA水平（ 原位杂交），或在提取核酸芯片或...

<table border="1"><tbody><tr><td> 物料 </td><td> 类型 </td><td> 公司 </td><td> 目录编号 </td></tr><tr><td> 昆虫文化维护材料 </td><td></td><td></td><td></td></tr><tr><td>酵母</td><td>活性干酵母</td><td> Fisher Scientific则</td><td> S802453 </td></tr><tr><td>飞食品</td><td>玉米粉，糖</td><td></td><td>标准食谱</td></tr><tr><td>蜂蜜</td><td>三叶草</td><td>荷兰黄金</td><td></td></tr><tr><td>样品瓶</td><td>聚丙烯壳样品瓶（窄） </td><td> Fisher Scientific则</td><td> AS514 </td></tr><tr><td>瓶封</td><td>棉塞</td><td> Fisher Scientific则</td><td> AS212 </td></tr><tr><td>瓶封</td><td>嗡嗡声插头</td><tD&gt;杰纳西科学</td><td> AS273 </td></tr><tr><td>冷藏孵化器</td><td>精密815 </td><td> Thermo Scientific的</td><td> 3721 </td></tr><tr><td></td><td></td><td></td><td></td></tr><tr><td> 材料样品制备 </td><td></td><td></td><td></td></tr><tr><td> CO 2坦克</td><td>骨干级</td><td>台湾史密斯</td><td> UN1013 </td></tr><tr><td>抹刀</td><td>微型抹刀（14厘米） </td><td> Fisher Scientific则</td><td> 21-401-15 </td></tr><tr><td> Pyrex的现场测试板</td><td> 9解剖钢板85毫米x 100毫米</td><td>托马斯科学</td><td> 7812G17 </td></tr><tr><td>巴斯德滴管</td><td>苏打石灰</td><td> J和Ĥ贝格</td><td> 71-5200-05 </td></tr><tr><td>镊子</td><td>风格＃5 </td><td>西格玛</td><td>的T-4662 </td></tr><tr><td>乙醇</td><td> 190证明美国药典</td><td> Fisher Scientific则</td><td> 04-355-221 </td></tr><tr><td>甲醛</td><td> 37％W / W </td><td> Fisher Scientific则</td><td> F79-1 </td></tr><tr><td>二级抗体Cy3标记AffiniPure驴抗兔IgG（H + L） </td><td> 1:50励磁546纳米;发射565纳米</td><td>杰克逊免疫研究实验室公司</td><td> 711-165-152 </td></tr><tr><td>抗淬灭（N - 没食子酸丙酯） </td><td> 4微克/毫升50％甘油在1X PBS </td><td> MP的Biomedicals </td><td> 10274790 </td></tr><tr><td>甘油</td><td></td><td> Fisher Scientific则</td><td> G33-1 </td></tr><tr><td>赫斯特33258 </td><td> 0.2微克/毫升激发352 nm的发射461纳米</td><td>分子探针</td><td>的H-1398 </td></tr><tr><td>罗丹明鬼笔环肽</td><td> 200个单位/毫升（6.6μM的）激发540 nm处，发射565纳米</td><td>分子探针</td><td> R415 </td></tr><tr><td>的Alexa Fluor 488鬼笔环肽</td><td> 300单位/毫升激发495 nm的发射518纳米</td><td>分子探针</td><td> A12379 </td></tr><tr><td></td><td></td><td></td><td></td></tr><tr><td> 耗材 </td><td></td><td></td><td></td></tr><tr><td>洗瓶</td><td> fisherbrand </td><td> Fisher Scientific则</td><td> 03-409-22A </td></tr><tr><td> kimwipes </td><td>金佰利</td><td> Fisher Scientific则</td><td> 06-666A </td></tr><tr><td>纸巾</td><td> 1层的C倍</td><td>鹅毛笔</td><td> 901-7CFTB2400 </td></tr><tr><td></td><td></td><td></td><td></td></tr><tr><td> 显微镜 </td><td> ＆NBSP; </td><td></td><td></td></tr><tr><td>徕卡立体显微镜</td><td> MZFLIII </td><td>帝国成像系统公司</td><td> 10446208 </td></tr><tr><td>蔡司体视显微镜</td><td> STEMI 1000或2000-C的</td><td>卡尔·蔡司</td><td> 000000-1006-126 </td></tr><tr><td>光源 - LED </td><td>鹅颈灯</td><td> Fisher Scientific则</td><td> 12563501 </td></tr><tr><td>阶段</td><td>钢板传输灯箱</td><td>卡尔·蔡司</td><td> 455137000 </td></tr><tr><td>蔡司激光扫描共聚焦显微镜</td><td> LSM的510 </td><td>卡尔·蔡司</td><td></td></tr><tr><td>蔡司复合显微镜</td><td> axioplan 2直立</td><td>卡尔·蔡司</td><td></td></tr></tbody></table><table border="1"><tbody><tr><td> 黄蜂株 </td><td> 飞应变小号</td></tr><tr><td> leptopilina victoriae 16 </td><td> YW </td></tr><tr><td> leptopilina boulardi 17 1 </td><td> UAS-GFP的背 17 </td></tr><tr><td> leptopilina heterotoma 2 </td><td> SerpentHemoGal4 13 </td></tr><tr><td> leptopilina heterotoma 14 1 </td><td> MSNF9-14 moCherry </td></tr><tr><td> trichopria果蝇 </td><td> MSNF-GFP的 15 </td></tr><tr><td> ganaspis xanthopoda 18 </td><td> YW蛇Gal4的无人机GFP-Dorsal/Basc 4 </td></tr><tr><td></td><td> YW; Drosomycin-GFP/CyO Y + 12 </td></tr></tbody></table>

an introduction to parasitic wasps of drosophila and the antiparasite immune response

最知名的寄生蜂攻击的果蝇幼虫或蛹的阶段。虽然Trichopria果蝇感染主机（ 图1A-C的 ），蛹的阶段，女性的，属Leptopilina（ 图1D，1F，1G）和Ganaspis（ 图1E）攻击的幼虫。我们使用这些寄生虫，一个种族的生物武器研究的分子基础。寄生蜂有防菌的巨大价值。他们大多携带的毒力和其他因素，修改主机的生理和免疫力。分析果蝇黄蜂提供特定物种的相互作用如何塑造自然群落的遗传结构的见解。这些研究还为了解主机的免疫生理以及如何协调免疫反应，这一类寄生虫挫败模型。 幼虫/蛹的表皮，作为第一线的夏德芬SE。蜂产卵器是一个尖锐的针状结构，可有效提供主机hemocoel的鸡蛋。其次是在表皮伤口愈合反应（ 图1C，箭头）产卵。有些黄蜂可以插入同一台主机上的两个或两个以上的鸡蛋，虽然只有一个鸡蛋的开发成功。的编外鸡蛋或发展幼虫被淘汰一个尚未了解的过程。这些小蜂，因此被称为为孤寄生。 根据动态应变和蜂种，蜂卵有两种命运。它可以封装，以便其发展受阻（主机出现; 图2左）;或黄蜂卵孵化，开发，molts，生长到成年（黄蜂出现图2右）。L。 heterotoma是果蝇寄生蜂的最好研究的物种之一。这是一个“多面手”，这意味着它可以利用大多数果蝇小号的pecies作为主机1 与heterotoma和L。 victoriae是姐妹物种和它们产生的病毒样颗粒封装响应2，积极干预。与L。 heterotoma，L. boulardi是一个专业的寄生虫和果蝇的范围，它采用的是相对有限的1。株L。 boulardi也产生病毒样颗粒3，虽然他们在自己的能力成功地在 D显着不同黑 1。一些这些研究boulardi株是很难成长， 在 D动态主机1 果蝇作为经常成功地封装它们的卵。因此，重要的是要在特定的实验协议双方的合作伙伴的知识。 此外屏障组织（角质层，肠道和气管）， 果蝇幼虫有全身的细胞免疫和体液免疫反应，产生fROM功能的血细胞和脂肪体，分别。产卵由L. boulardi同时激活免疫武器1,4。流通中的血细胞被发现，在柄的人群，分段角质层下和淋巴腺。淋巴腺是一个小的背侧幼虫的造血器官。造血干细胞群，称为叶，成对排列分段沿背船只沿着动物的前后轴（ 图3A）运行。脂肪体是一个大型的多功能器官（ 图3B）。它分泌抗菌肽微生物和后生动物感染。 黄蜂感染激活免疫信号（ 图4）4。在细胞水平上，它触发的血细胞的分裂和分化。在自卫，聚集和胶囊开发感染动物hemocoel（ 图5）5,6。 ACtivated血细胞迁移向蜂卵（或黄蜂幼虫）和它周围开始形成一个胶囊（ 图5A - F）。一些血细胞聚集形成结节（ 图5G - H）。仔细分析发现，黄蜂感染引起淋巴腺叶最前，在其外围分散（ 图6C，D）的。 我们提出用电话信号转导通路元件背和Spätzle（ 图4,5,7），其目标Drosomycin（ 图6）具有代表性的数据，来说明如何在淋巴腺和hemocoel的具体变化，可以研究黄蜂感染后。这里描述的剥离协议，也能产生从主机淋巴黄蜂（ 图8）鸡蛋（或发展的黄蜂阶段）。

Watch this Scientific Journal Video about An Introduction to Parasitic Wasps of Drosophila and the Antiparasite Immune Response at JoVE.com

An Introduction to Parasitic Wasps of Drosophila and the Antiparasite Immune Response

寄生蜂（寄生）构成了许多昆虫的天敌，包括各大类<em&gt;果蝇</em&gt;。我们将引进的技术，这些寄生虫传播<em&gt;果蝇</em&gt;属。并演示如何分析免疫组织的影响<em&gt;果蝇</em&gt;幼虫。

寄生蜂简介<em&gt;果蝇</em&gt;和Antiparasite的免疫反应的

Most known parasitoid wasp species attack the larval or pupal stages of Drosophila. While Trichopria drosophilae infect the pupal stages of the host (Fig. ...

an-introduction-to-parasitic-wasps-drosophila-antiparasite-immune

Research

JoVE Journal

Immunology and Infection

An Introduction to Parasitic Wasps of Drosophila and the Antiparasite Immune Response

18.4K 次观看.  The City College of New York, CUNY. 寄生蜂（寄生）构成了许多昆虫的天敌，包括各大类果蝇。我们将引进的技术，这些寄生虫传播果蝇属。并演示如何分析免疫组织的影响果蝇幼虫。

视频: An Introduction to Parasitic Wasps of Drosophila and the Antiparasite Immune Response

Multicolor Flow Cytometry Analyses of Cellular Immune Response in Rhesus Macaques

The rhesus macaque model is currently the best available model for HIV-AIDS with respect to understanding the pathogenesis as well as for the development of vaccines and therapeutics1,2,3. Here, we describe a method for the detailed phenotypic and functional analyses of cellular immune responses, specifically intracellular cytokine production by CD4+ and CD8+ T cells as well as the individual memory subsets. We obtained precise quantitative and qualitative measures for the production of interferon gamma (INF-) and interleukin (IL) -2 in both CD4+ and CD8+ T cells from the rhesus macaque PBMC stimulated with PMA plus ionomycin (PMA+I). The cytokine profiles were different in the different subsets of memory cells. Furthermore, this protocol provided us the sensitivity to demonstrate even minor fractions of antigen specific CD4+ and CD8+ T cell subsets within the PBMC samples from rhesus macaques immunized with an HIV envelope peptide cocktail vaccine developed in our laboratory. The multicolor flow cytometry technique is a powerful tool to precisely identify different populations of T cells 4,5 with cytokine-producing capability6 following non-specific or antigen-specific stimulation 5,7.

We demonstrate the utility of multicolor flow cytometry for detailed phenotypic and functional characterization of total as well as memory subsets of CD4+ and CD8+ T cells in rhesus macaques, the ideal model for HIV/AIDS vaccine studies.

多色流流式细胞仪分析细胞在恒河猴的免疫应答

The rhesus macaque model is currently the best available model for HIV-AIDS with respect to understanding the pathogenesis as well as for the development ...

科研

免疫与感染

Quantitative Measurement of the Immune Response and Sleep in Drosophila

A complex interaction between the immune response and host behavior has been described in a wide range of species. Excess sleep, in particular, is known to occur as a response to infection in mammals 1 and has also recently been described in Drosophila melanogaster2. It is generally accepted that sleep is beneficial to the host during an infection and that it is important for the maintenance of a robust immune system3,4. However, experimental evidence that supports this hypothesis is limited4, and the function of excess sleep during an immune response remains unclear. We have used a multidisciplinary approach to address this complex problem, and have conducted studies in the simple genetic model system, the fruitfly Drosophila melanogaster. We use a standard assay for measuring locomotor behavior and sleep in flies, and demonstrate how this assay is used to measure behavior in flies infected with a pathogenic strain of bacteria. This assay is also useful for monitoring the duration of survival in individual flies during an infection. Additional measures of immune function include the ability of flies to clear an infection and the activation of NF&kappa;B, a key transcription factor that is central to the innate immune response in Drosophila. Both survival outcome and bacterial clearance during infection together are indicators of resistance and tolerance to infection. Resistance refers to the ability of flies to clear an infection, while tolerance is defined as the ability of the host to limit damage from an infection and thereby survive despite high levels of pathogen within the system5. Real-time monitoring of NF&kappa;B activity during infection provides insight into a molecular mechanism of survival during infection. The use of Drosophila in these straightforward assays facilitates the genetic and molecular analyses of sleep and the immune response and how these two complex systems are reciprocally influenced.

Quantitative Measurement of the Immune Response and Sleep in Drosophila

To understand a link between the immune response and behavior, we describe a method to measure locomotor behavior in Drosophila during bacterial infection as well as the ability of flies to mount an immune response by monitoring survival, bacterial load, and real-time activity of a key regulator of innate immunity, NF&kappa;B.

定量测定的免疫反应和睡眠模式<em&gt;果蝇</em

A complex interaction between the immune response and  host behavior has been described in a wide range of species. Excess sleep, in  particular, is known ...

Quantification of the Respiratory Burst Response as an Indicator of Innate Immune Health in Zebrafish

The phagocyte respiratory burst is part of the innate immune response to pathogen infection and involves the production of reactive oxygen species (ROS). ROS are toxic and function to kill phagocytized microorganisms. In vivo quantification of phagocyte-derived ROS provides information regarding an organism's ability to mount a robust innate immune response. Here we describe a protocol to quantify and compare ROS in whole zebrafish embryos upon chemical induction of the phagocyte respiratory burst. This method makes use of a non-fluorescent compound that becomes fluorescent upon oxidation by ROS. Individual zebrafish embryos are pipetted into the wells of a microplate and incubated in this fluorogenic substrate with or without a chemical inducer of the respiratory burst. Fluorescence in each well is quantified at desired time points using a microplate reader. Fluorescence readings are adjusted to eliminate background fluorescence and then compared using an unpaired t-test. This method allows for comparison of the respiratory burst potential of zebrafish embryos at different developmental stages and in response to experimental manipulations such as protein knockdown, overexpression, or treatment with pharmacological agents. This method can also be used to monitor the respiratory burst response in whole dissected kidneys or cell preparations from kidneys of adult zebrafish and some other fish species. We believe that the relative simplicity and adaptability of this protocol will complement existing protocols and will be of interest to researchers who seek to better understand the innate immune response.

The innate immune response protects organisms against pathogen infection. A critical component of the innate immune response, the phagocyte respiratory burst, generates reactive oxygen species that kill invading microorganisms. We describe a respiratory burst assay that quantifies reactive oxygen species produced when the innate immune response is chemically induced.

呼吸爆发的响应为先天免疫健康的斑马鱼中的指标量化

The phagocyte respiratory burst is part of the innate immune response to  pathogen infection and involves the production of reactive oxygen species (ROS). ...

Investigating the Effects of Probiotics on Pneumococcal Colonization Using an In Vitro Adherence Assay

Adherence of Streptococcus pneumoniae (the pneumococcus) to the epithelial lining of the nasopharynx can result in colonization and is considered a prerequisite for pneumococcal infections such as pneumonia and otitis media. In vitro adherence assays can be used to study the attachment of pneumococci to epithelial cell monolayers and to investigate potential interventions, such as the use of probiotics, to inhibit pneumococcal&#160;colonization.&#160;The protocol described here is used to investigate the effects of the probiotic Streptococcus salivarius on the adherence of pneumococci to the human epithelial cell line CCL-23 (sometimes referred to as HEp-2 cells). The assay involves three main steps: 1) preparation of epithelial and bacterial cells, 2) addition of bacteria to epithelial cell monolayers, and 3) detection of adherent pneumococci by viable counts (serial dilution and plating) or quantitative real-time PCR (qPCR). This technique is relatively straightforward and does not require specialized equipment other than a tissue culture setup. The assay can be used to test other probiotic species and/or potential inhibitors of pneumococcal colonization and can be easily modified to address other scientific questions regarding pneumococcal adherence and invasion.

Investigating the Effects of Probiotics on Pneumococcal Colonization Using an In Vitro Adherence Assay

In vitro adherence assays can be used to study the attachment of Streptococcus pneumoniae to epithelial cell monolayers and to investigate potential interventions such as the use of probiotics for inhibiting pneumococcal colonization.

使用一个调查益生素对肺炎球菌定植的影响<em&gt;体外</em&gt;粘附试验

Adherence of Streptococcus pneumoniae (the pneumococcus) to the epithelial lining of the nasopharynx can result in colonization and is considered a ...

Using RNA-interference to Investigate the Innate Immune Response in Mouse Macrophages

Macrophages are key phagocytic innate immune cells. When macrophages encounter a pathogen, they produce antimicrobial proteins and compounds to kill the pathogen, produce various cytokines and chemokines to recruit and stimulate other immune cells, and present antigens to stimulate the adaptive immune response. Thus, being able to efficiently manipulate macrophages with techniques such as RNA-interference (RNAi) is critical to our ability to investigate this important innate immune cell. However, macrophages can be technically challenging to transfect and can exhibit inefficient RNAi-induced gene knockdown. In this protocol, we describe methods to efficiently transfect two mouse macrophage cell lines (RAW264.7 and J774A.1) with siRNA using the Amaxa Nucleofector 96-well Shuttle System and describe procedures to maximize the effect of siRNA on gene knockdown. Moreover, the described methods are adapted to work in 96-well format, allowing for medium and high-throughput studies. To demonstrate the utility of this approach, we describe experiments that utilize RNAi to inhibit genes that regulate lipopolysaccharide (LPS)-induced cytokine production.

In this protocol, we describe methods to efficiently transfect murine macrophage cell lines with siRNAs using the Amaxa Nucleofector 96-well Shuttle System, stimulate the macrophages with lipopolysaccharide, and monitor the effects on inflammatory cytokine production.

利用RNA干扰调查的先天免疫反应的小鼠巨噬细胞

Macrophages are key phagocytic innate immune cells.  When macrophages encounter a pathogen, they produce antimicrobial proteins and compounds to kill the ...

The Utilization of Oropharyngeal Intratracheal PAMP Administration and Bronchoalveolar Lavage to Evaluate the Host Immune Response in Mice

The host immune response to pathogens is a complex biological process. The majority of in vivo studies classically employed to characterize host-pathogen interactions take advantage of intraperitoneal injections of select bacteria or pathogen associated molecular patterns (PAMPs) in mice. While these techniques have yielded tremendous data associated with infectious disease pathobiology, intraperitoneal injection models are not always appropriate for host-pathogen interaction studies in the lung. Utilizing an acute lung inflammation model in mice, it is possible to conduct a high resolution analysis of the host innate immune response utilizing lipopolysaccharide (LPS). Here, we describe the methods to administer LPS using nonsurgical oropharyngeal intratracheal administration, monitor clinical parameters associated with disease pathogenesis, and utilize bronchoalveolar lavage fluid to evaluate the host immune response. The techniques that are described are widely applicable for studying the host innate immune response to a diverse range of PAMPs and pathogens. Likewise, with minor modifications, these techniques can also be applied in studies evaluating allergic airway inflammation and in pharmacological applications.

The host immune response to pathogen infection is a tightly regulated process. Utilizing a lipopolysaccharide lung exposure model in mice, it is possible to conduct high resolution evaluations of the complex mechanisms associated with disease pathogenesis.

口咽部气管内PAMP管理和支气管肺泡灌洗的利用率来评估小鼠宿主的免疫反应

The host immune response to pathogens is a complex biological process. The majority of in vivo studies classically employed to characterize host-pathogen ...

In Vitro Analysis of Myd88-mediated Cellular Immune Response to West Nile Virus Mutant Strain Infection

An attenuated West Nile virus (WNV), a nonstructural (NS) 4B-P38G mutant, induced higher innate cytokine and T cell responses than the wild-type WNV in mice. Recently, myeloid differentiation factor 88 (MyD88) signaling was shown to be important for initial T cell priming and memory T cell development during WNV NS4B-P38G mutant infection. In this study, two flow cytometry-based methods &#8211; an in vitro T cell priming assay and an intracellular cytokine staining (ICS) &#8211; were utilized to assess dendritic cells (DCs) and T cell functions. In the T cell priming assay, cell proliferation was analyzed by flow cytometry following co-culture of DCs from both groups of mice with carboxyfluorescein succinimidyl ester (CFSE) - labeled CD4+ T cells of OTII transgenic mice. This approach provided an accurate determination of the percentage of proliferating CD4+ T cells with significantly improved overall sensitivity than the traditional assays with radioactive reagents. A microcentrifuge tube system was used in both cell culture and cytokine staining procedures of the ICS protocol. Compared to the traditional tissue culture plate-based system, this modified procedure was easier to perform at biosafety level (BL) 3 facilities. Moreover, WNV- infected cells were treated with paraformaldehyde in both assays, which enabled further analysis outside BL3 facilities. Overall, these in vitro immunological assays can be used to efficiently assess cell-mediated immune responses during WNV infection.

In Vitro Analysis of Myd88-mediated Cellular Immune Response to West Nile Virus Mutant Strain Infection

Two flow cytometry-based methods &#8211; an in vitro T cell priming assay and intracellular cytokine staining were utilized to measure antigen presenting capacity of dendritic cells and antigen-specific T cell responses to a West Nile virus mutant infection in mice.

在 MyD88的介导的细胞免疫应答西尼罗河病毒变异株感染体外分析

An attenuated West Nile virus (WNV), a nonstructural (NS) 4B-P38G mutant, induced higher innate cytokine and T cell responses than the wild-type WNV in ...

Non-invasive Imaging of the Innate Immune Response in a Zebrafish Larval Model of Streptococcus iniae Infection

The aquatic pathogen, Streptococcus iniae, is responsible for over 100 million dollars in annual losses for the aquaculture industry and is capable of causing systemic disease in both fish and humans. A better understanding of S. iniae disease pathogenesis requires an appropriate model system. The genetic tractability and the optical transparency of the early developmental stages of zebrafish allow for the generation and non-invasive imaging of transgenic lines with fluorescently tagged immune cells. The adaptive immune system is not fully functional until several weeks post fertilization, but zebrafish larvae have a conserved vertebrate innate immune system with both neutrophils and macrophages. Thus, the generation of a larval infection model allows the study of the specific contribution of innate immunity in controlling S. iniae infection.
The site of microinjection will determine whether an infection is systemic or initially localized. Here, we present our protocols for otic vesicle injection of zebrafish aged 2-3 days post fertilization as well as our techniques for fluorescent confocal imaging of infection. A localized infection site allows observation of initial microbe invasion, recruitment of host cells and dissemination of infection. Our findings using the zebrafish larval model of S. iniae infection indicate that zebrafish can be used to examine the differing contributions of host neutrophils and macrophages in localized bacterial infections. In addition, we describe how photolabeling of immune cells can be used to track individual host cell fate during the course of infection.

Non-invasive Imaging of the Innate Immune Response in a Zebrafish Larval Model of Streptococcus iniae Infection

Here, we present a protocol for the generation and imaging of a localized bacterial infection in the zebrafish otic vesicle.

先天免疫反应中的斑马鱼幼体型号无创成像<em&gt;链球菌iniae</em&gt;感染

The aquatic pathogen, Streptococcus iniae, is responsible for over 100 million dollars in annual losses for the aquaculture industry and is capable of ...

An In Vitro Model for Measuring Immune Responses to Malaria in the Context of HIV Co-infection

Malaria and HIV co-infection is a growing health priority. However, most research on malaria or HIV currently focuses on each infection individually. Although understanding the disease dynamics for each of these pathogens independently is vital, it is also important that the interactions between these pathogens are investigated and understood. 
We have developed a versatile in vitro model of HIV-malaria co-infection to study host immune responses to malaria in the context of HIV infection. Our model allows the study of secreted factors in cellular supernatants, cell surface and intracellular protein markers, as well as RNA expression levels. The experimental design and methods used limit variability and promote data reliability and reproducibility. 
All pathogens used in this model are natural human pathogens (Plasmodium falciparum and HIV-1), and all infected cells are naturally infected and used fresh. We use human erythrocytes parasitized with P. falciparum and maintained in continuous in vitro culture. We obtain freshly isolated peripheral blood mononuclear cells from chronically HIV-infected volunteers. Every condition used has an appropriate control (P. falciparum parasitized vs. normal erythrocytes), and every HIV-infected donor has an HIV uninfected control, from which cells are harvested on the same day. This model provides a realistic environment to study the interactions between malaria parasites and human immune cells in the context of HIV infection.

An In Vitro Model for Measuring Immune Responses to Malaria in the Context of HIV Co-infection

Human co-infection is difficult to replicate in vitro. However, human malaria parasites can readily be cultured in vitro, as can freshly isolated human peripheral blood mononuclear cells naturally infected with HIV. This provides an excellent model for studying early immune responses to malaria parasites in the context of HIV co-infection.

一个<em&gt;体外</em&gt;模型在HIV合并感染的背景下测量免疫应答疟疾

Malaria and HIV co-infection is a growing health priority. However, most research on malaria or HIV currently focuses on each infection individually. ...

Inoculating Anopheles gambiae Mosquitoes with Beads to Induce and Measure the Melanization Immune Response

The stimulation of immune responses is a common tool in invertebrate studies to examine the efficacy and the mechanisms of immunity. This stimulation is based on the injection of non-pathogenic particles into insects, as the particles will be detected by the immune system and will induce the production of immune effectors. We focus here on the stimulation of the melanization response in the mosquito Anopheles gambiae. The melanization response results in the encapsulation of foreign particles and parasites with a dark layer of melanin. To stimulate this response, mosquitoes are inoculated with beads in the thoracic cavity using microcapillary glass tubes. Then, after 24 hr, the mosquitoes are dissected to retrieve the beads. The degree of melanization of the bead is measured using image analysis software. Beads do not have the pathogenic effects of parasites, or their capacity to evade or suppress the immune response. These injections are a way to measure immune efficacy and the impact of immune stimulations on other life history traits, such as fecundity or longevity. It is not exactly the same as directly studying host-parasite interactions, but it is an interesting tool to study immunity and its evolutionary ecology.

Inoculating Anopheles gambiae Mosquitoes with Beads to Induce and Measure the Melanization Immune Response

Through inoculation with beads, the described technique enables the stimulation of the mosquito melanization response in the hemolymph circulating system. The amount of melanin covering the beads can be measured after dissection as a measure of the immune response.

接种<em&gt;冈比亚按蚊</em&gt;珠蚊子诱发和测量黑化免疫反应

The stimulation of immune responses is a common tool in invertebrate studies to examine the efficacy and the mechanisms of immunity. This stimulation is ...