Name: measuring phagocytosis of aspergillus fumigatus conidia by human leukocytes using flow cytometry
Uploaded: 2019-12-07T12:00:00
Description: Watch this Scientific Journal Video about Measuring Phagocytosis of Aspergillus fumigatus Conidia by Human Leukocytes using Flow Cytometry at JoVE.com

我们感谢皮娅·斯蒂尔夫人的出色技术援助。M. von Lilienfeld-Toal 由败血症控制和护理中心（德国联邦教育和卫生部、BMBF、FKZ 01E01002）和感染性生物研究园区（BMBF，FKZ 13GW0096D）提供支持。蒂恩戈克·迈·霍恩由耶拿微生物传播学院（德国福森斯格明舍夫特，FKZ 214/2）支持

该协议包括使用从输血医学研究所、耶拿大学医院获得的人类牛皮大衣和从患者身上抽取的新鲜静脉血，均需经献血者书面知情同意，符合伦理委员会批准书4357-03/15。
1.阿斯珀吉鲁斯熏蒸菌的制备
<ol><li>在37°C下，在37°C下种植A.熏蒸，在1.5%麦芽阿加培养皿上5天，不含CO2。 注意：熏蒸是生物安全2级微生物，必须在适当的设施中使?...

<ol>
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F., Behnsen, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Interaction+of+phagocytes+with+filamentous+fungi.">Interaction of phagocytes with filamentous fungi.</a> Current Opinion in Microbiology. 13 (4), 409-415 (2010).</li><li>Heinekamp, 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=Interference+of+Aspergillus+fumigatus+with+the+immune+response.">Interference of Aspergillus fumigatus with the immune response.</a> Seminars in Immunopathology. 37 (2), 141-152 (2015).</li><li>Slesiona, S., 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=Persistence+versus+escape:+Aspergillus+terreus+and+Aspergillus+fumigatus+employ+different+strategies+during+interactions+with+macrophages.">Persistence versus escape: Aspergillus terreus and Aspergillus fumigatus employ different strategies during interactions with macrophages.</a> PLoS One. 7 (2), 31223 (2012).</li><li>Busetto, S., Trevisan, E., Patriarca, P., Menegazzi, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+single-step,+sensitive+flow+cytofluorometric+assay+for+the+simultaneous+assessment+of+membrane-bound+and+ingested+Candida+albicans+in+phagocytosing+neutrophils.">A single-step, sensitive flow cytofluorometric assay for the simultaneous assessment of membrane-bound and ingested Candida albicans in phagocytosing neutrophils.</a> Cytometry A. 58 (2), 201-206 (2004).</li><li>Lowe, D. 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M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Flow+cytometric+quantitative+determination+of+ingestion+by+phagocytes+needs+the+distinguishing+of+overlapping+populations+of+binding+and+ingesting+cells.">Flow cytometric quantitative determination of ingestion by phagocytes needs the distinguishing of overlapping populations of binding and ingesting cells.</a> Cytometry A. 65 (2), 93-102 (2005).</li><li>Saresella, 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+rapid+evaluation+of+phagocytosis+and+killing+of+Candida+albicans+by+CD13++leukocytes.">A rapid evaluation of phagocytosis and killing of Candida albicans by CD13+ leukocytes.</a> Journal of Immunological Methods. 210 (2), 227-234 (1997).</li><li>Fattorossi, A., Nisini, R., Pizzolo, J. G., D'Amelio, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=New,+simple+flow+cytometry+technique+to+discriminate+between+internalized+and+membrane-bound+particles+in+phagocytosis.">New, simple flow cytometry technique to discriminate between internalized and membrane-bound particles in phagocytosis.</a> Cytometry. 10 (3), 320-325 (1989).</li><li>Heinzelmann, M., Gardner, S. A., Mercer-Jones, M., Roll, A. J., Polk, H. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Quantification+of+phagocytosis+in+human+neutrophils+by+flow+cytometry.">Quantification of phagocytosis in human neutrophils by flow cytometry.</a> Microbiology and Immunology. 43 (6), 505-512 (1999).</li><li>Perticarari, S., Presani, G., Mangiarotti, M. A., Banfi, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Simultaneous+flow+cytometric+method+to+measure+phagocytosis+and+oxidative+products+by+neutrophils.">Simultaneous flow cytometric method to measure phagocytosis and oxidative products by neutrophils.</a> Cytometry. 12 (7), 687-693 (1991).</li><li>Sveum, R. J., Chused, T. M., Frank, M. M., Brown, E. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+quantitative+fluorescent+method+for+measurement+of+bacterial+adherence+and+phagocytosis.">A quantitative fluorescent method for measurement of bacterial adherence and phagocytosis.</a> Journal of Immunolological Methods. 90 (2), 257-264 (1986).</li><li>de Boer, E. C., Bevers, R. F., Kurth, K. H., Schamhart, D. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Double+fluorescent+flow+cytometric+assessment+of+bacterial+internalization+and+binding+by+epithelial+cells.">Double fluorescent flow cytometric assessment of bacterial internalization and binding by epithelial cells.</a> Cytometry. 25 (4), 381-387 (1996).</li><li>Hartung, S., 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=Fast+and+Quantitative+Evaluation+of+Human+Leukocyte+Interaction+with+Aspergillus+fumigatus+Conidia+by+Flow+Cytometry.">Fast and Quantitative Evaluation of Human Leukocyte Interaction with Aspergillus fumigatus Conidia by Flow Cytometry.</a> Cytometry A. 95 (3), 332-338 (2019).</li><li>Fei, C., Lillico, D. M. E., Hall, B., Rieger, A. M., Stafford, J. 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该协议提出了一种快速流动细胞学方法，用于测量A.熏蒸锥体与大量初级人类白细胞的相互作用，这在常见的显微协议中是不可能的。使用显微镜和手动计数内化锥体成像细胞非常麻烦，实际上只能对几百个细胞进行成像。流式细胞测量通过在几分钟内测量数千个细胞来克服这个问题。这两种方法共有的障碍是细胞内或细胞上的邻形同质体的区别。在显微镜中，染料钙氟白色常用于染色粘?...

侵入性肺动脉粥样硬化对免疫功能低下患者是一个很大的威胁，因为治疗方案有限，只有在早期诊断成功，导致高死亡率1。传染性病原体是阿塞珀吉鲁斯熏蒸剂的锥度（孢子），在大多数栖息地无处不在2。Conidia 被吸入，通过气道，最后可以进入肺肺泡。在免疫能力人类中，这些锥体由先天免疫细胞清除，如单核细胞或巨噬细胞和嗜中性粒细胞，它们吸收（噬菌体）并消化病原体3。当对宿主-病原体相互作用感兴趣时，噬菌体病对微生物学家和免疫学家也很重要。对抗测定，如白细胞和锥体的共同孵育，通常包括用荧光素或其衍生的荧光素等子素（FITC）标记孢子。使用显微镜，它很容易识别内化荧光锥体，并确定附加/粘附锥体，虽然这种方法是繁琐和现实地限于几百个细胞4。然而，在流式细胞学中，很容易在几分钟内分析数十万个细胞，对方位细胞和粘附性锥体的不同染色至关重要。因此，许多协议依靠Trypan Blue来淬火FITC荧光从粘附锥体5，6，7，8。另一种方法是利用溴化和FITC的荧光共振能量转移，从粘附的锥度9，10，11发射红色，而不是绿色荧光。如果有特定的抗体可用，就像某些细菌一样，细胞结合的颗粒可以直接染色12，13。
在这里，我们提出一个协议，通过采用异位素青霉素（APC）耦合抗FITC抗体，快速定量地评估带有PCC标签的人类白细胞的芬加菌锥体，以及孢子附着在细胞上和缺乏相互作用。该方法还允许同时对来自同一样品的单核细胞和嗜中性粒细胞的食细胞化进行进一步细胞标记的流动细胞学分析。
该协议可应用于真菌菌株的表征（例如，这里介绍的Mucorales属的几种Aspergillus和其他霉菌）及其突变体14和对噬菌体（如免疫功能低下个体的白细胞）的免疫学研究。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Adhesive foil</td>
			<td>Brand</td>
			<td>701367</td>
			<td></td>
		</tr>
		<tr>
			<td>anti-CD14 V500</td>
			<td>BD Biosciences</td>
			<td>561391</td>
			<td>clone M5E2</td>
		</tr>
		<tr>
			<td>anti-CD45 BUV395</td>
			<td>BD Biosciences</td>
			<td>563792</td>
			<td>clone HI30</td>
		</tr>
		<tr>
			<td>anti-CD66b PerCP-Cy5.5</td>
			<td>BD Biosciences</td>
			<td>562254</td>
			<td>clone G10F5</td>
		</tr>
		<tr>
			<td>anti-FITC APC</td>
			<td>ThermoFisher Scientific</td>
			<td>17-7691-82</td>
			<td>clone NAWESLEE</td>
		</tr>
		<tr>
			<td>Cell culture plate, 12-well</td>
			<td>Greiner Bio-one</td>
			<td>665180</td>
			<td></td>
		</tr>
		<tr>
			<td>Cell scraper</td>
			<td>Bioswisstech</td>
			<td>800020</td>
			<td></td>
		</tr>
		<tr>
			<td>Cell strainer, 30 &#181;m</td>
			<td>Miltenyi Biotech</td>
			<td>130-098-458</td>
			<td>SmartStrainer</td>
		</tr>
		<tr>
			<td>Cytometer</td>
			<td>BD Biosciences</td>
			<td></td>
			<td>LSR Fortessa II, lasers: 488 nm (blue), 405 nm (violet), 355 nm (UV) and 640 nm (red)</td>
		</tr>
		<tr>
			<td>Detergent</td>
			<td>Sigma Aldrich</td>
			<td>P1379</td>
			<td>Tween 20, 0.01% in PBS</td>
		</tr>
		<tr>
			<td>Drigalski spatula</td>
			<td>Carl Roth</td>
			<td>PC59.1</td>
			<td></td>
		</tr>
		<tr>
			<td>Ethylenediaminetetraacetic acid (EDTA)</td>
			<td>Sigma Aldrich</td>
			<td>ED3SS-500g</td>
			<td>2 mM in PBS</td>
		</tr>
		<tr>
			<td>Erythrocyte lysis buffer</td>
			<td></td>
			<td></td>
			<td>0.15 M NH4Cl, 10 mM KHCO3, 0.1 mM EDTA</td>
		</tr>
		<tr>
			<td>Fetal Calf Serum (FCS)</td>
			<td>Biochrom AG</td>
			<td>S 0115</td>
			<td>10% in RPMI 1640</td>
		</tr>
		<tr>
			<td>Fluorescein isothiocyanate (FITC)</td>
			<td>Sigma Aldrich</td>
			<td>F3651-100MG</td>
			<td>0.1 mM in Na2CO3 /PBS solution</td>
		</tr>
		<tr>
			<td>Formaldehyd</td>
			<td>Carl Roth</td>
			<td>PO87.3</td>
			<td>Histofix</td>
		</tr>
		<tr>
			<td>Malt agar (1.5%)</td>
			<td></td>
			<td></td>
			<td>malt extract (40 g), yeast extract (4 g), agar (15 g), Aqua dest. (1 L), adjust pH to 5.7-6.0, sterilise at 121 &#176;C for 35 minutes</td>
		</tr>
		<tr>
			<td>Na2CO3</td>
			<td>Carl Roth</td>
			<td>8563.1</td>
			<td>0.1 M in PBS</td>
		</tr>
		<tr>
			<td>Petri dish</td>
			<td>Greiner Bio-one</td>
			<td>633180</td>
			<td></td>
		</tr>
		<tr>
			<td>Phosphat Buffered Saline (PBS)</td>
			<td>ThermoFisher Scientific</td>
			<td>189012-014</td>
			<td>without Calcium, without Magnesium</td>
		</tr>
		<tr>
			<td>RPMI 1640</td>
			<td>ThermoFisher Scientific</td>
			<td>61870010</td>
			<td>RPMI 1640 Medium, GlutaMAX Supplement</td>
		</tr>
		<tr>
			<td>Rotator</td>
			<td>Miltenyi Biotech</td>
			<td>130-090-753</td>
			<td>MACSmix Tube Rotator</td>
		</tr>
		<tr>
			<td>Round-bottom tube, 7.5 mL</td>
			<td>Corning</td>
			<td>REF 352008</td>
			<td></td>
		</tr>
		<tr>
			<td>Software for data acquisition and analysis</td>
			<td>BD Biosciences</td>
			<td></td>
			<td>FACSDiva 8.0</td>
		</tr>
		<tr>
			<td>V-bottom plate, 96 well</td>
			<td>Brand</td>
			<td>781601</td>
			<td>untreated surface</td>
		</tr>
	</tbody>
</table>

measuring phagocytosis of aspergillus fumigatus conidia by human leukocytes using flow cytometry

霉菌阿斯珀吉鲁斯熏蒸菌的侵入性肺感染对免疫功能低下的患者构成巨大威胁。吸入真菌锥体（孢子）通过先天单核细胞和/或中性粒细胞的噬菌体从人类肺肺泡。该协议提供了一种快速、可靠的测量方细胞化，使用氟素等氰酸酯 （FITC） 标记的锥体与人类白细胞共同孵育，随后与抗 FITC 抗体进行反染色，以便对内化和细胞粘附锥体进行鉴别。该协议的主要优点是其快速性，将测定与其他感兴趣的细胞标记的细胞学分析相结合的可能性，同时分析单个样本中的单核细胞和嗜中性粒细胞，以及其对其他细胞壁携带真菌或细菌的适用性。测定方位白细胞的百分比，为微生物学家评估病原体的毒性或比较病原体的野生型和突变体提供了手段，也为免疫学家提供了研究人类白细胞对抗病原体的能力的手段。

在人类吞噬细胞测量锥体噬菌体吞噬细胞时，真正的内化和仅仅依附于细胞之间的歧视是一个障碍，特别是在流式细胞测定等高通量方法方面。为了克服这一障碍，我们提出了一种快速可靠的方案，基于在细胞和锥体联合孵育之前用荧光染料FITC染色的锥体，然后与APC标记的抗FITC抗体在孵育后进行反染色（图1A）。如图1B...

Watch this Scientific Journal Video about Measuring Phagocytosis of Aspergillus fumigatus Conidia by Human Leukocytes using Flow Cytometry at JoVE.com

Measuring Phagocytosis of Aspergillus fumigatus Conidia by Human Leukocytes using Flow Cytometry

该协议提供了一种快速可靠的方法，用流动细胞测定人类原发性噬菌体定量测量阿苯甲基的邻苯二甲苯，并区分锥体从单纯的粘附到白细胞的邻形图。

利用流细胞测定人类白细胞测定阿斯珀吉鲁斯紫锥体烟曲的噬菌体

Invasive pulmonary infection by the mold Aspergillus fumigatus poses a great threat to immunocompromised patients. Inhaled fungal conidia (spores) are ...

该协议意义重大，因为它定量测量人类白细胞标记的阿斯佩吉卢斯熏蒸器康尼迪亚的噬菌体，以及康尼迪亚与细胞的附着，以及流式细胞学缺乏相互作用。这种方法的主要优点是，它便于快速分析大量的细胞。此外，细胞标记的标签允许在同一样本中分离对嗜中性粒细胞和单核细胞的评估。在任何这种技术可用于分析其他临床相关的真菌，如木瓜。相反，红血球可能被用作噬菌体。要开始此过程，请用消毒剂将纸巾弄湿，然后放入生物安全柜中。将一盘生长的 Aspergillus fumigatus 放在纸巾上，以防止挥发性 Conidia 过度分布。在真菌上加入含有0.01%洗涤剂的10毫升PBS，并使用Drigalski铲将液体铺在盘子上，擦掉深色的康迪迪亚。通过 30 微米过滤器将 Conidia 悬浮液转移到 50 毫升，这将去除任何残留的 mycelia。再加入10毫升的PBS，用洗涤剂加入板中，用铲子铺开，然后通过过滤器将其转移到同一管中。接下来，准备0.1摩尔碳酸钠的无菌溶液，溶解在PBS中。并在此碳酸钠溶液中准备0.1毫摩尔菲特粉末溶液。在此 FITC 溶液的 5 毫升中，在 15 毫升管中重新悬浮 1 亿或更少的 Conidia。在37摄氏度的旋转器中孵育20分钟。要膨胀到康迪迪亚，重新暂停在五毫升的 RPMI 介质中标记的 FITC 标记的 Conidia，并辅以 10%FCS。并在37摄氏度的旋转器中孵育所需的时间。在生物安全柜中，将五毫升的布菲大衣放入50毫升的管子中。用EL缓冲液填充管子，并反转三次。水平孵育五至八分钟，直到混合物的乳白色外观变清。然后，在300倍G下离心，在室温下10分钟。丢弃上流液，通过移液将颗粒重新在一毫升 EL 缓冲液中。再加入24毫升EL缓冲液，并多次反转管子。在300倍G下离心，在室温下离心5分钟。丢弃上一提液，将细胞重新在一毫升的RPMI介质中，辅以10%FCS。首先，将200万白细胞和400万个标有 FITC 的 Conidia 在 1.5 毫升的 RPMI 中，辅以 10%FCS，转移到 12 孔培养板。包括仅对没有 Conidia 的单元格进行控制，以及对未标记的 Conidia 的细胞的控制。在加湿的二氧化碳培养箱中孵育板，温度为37摄氏度，达到所需时间。当孵育完成时，使用细胞刮刀收集细胞，并将它们转移到15毫升管。首先，将每个样品的 100 微升和控制装置放入 96 井 V 底板的一个井中。加入 150 微升 PBS，含有 2 毫摩尔 EDTA 洗涤。对于颜色补偿，将一百万个无 Conidia 的细胞放在板的其他孔中。包括一个将不受约束的细胞井。在每个井中加入 150 微升 PBS，其中含有 2 毫摩尔的 EDTA。接下来，用粘合箔盖住板，在300倍G和室温下离心5分钟。然后，取出铝箔，快速、有力地将盘子倒置一次，或在水槽或一次性纸巾上丢弃上一遍。将细胞重新在100微升抗体混合物中，通过移液混合良好。对于颜色补偿，在含有两毫升EDTA的100微升PBS中重新填充各自的细胞，并在每个井中加入一种抗体类型，其量与抗体混合物中使用的相同。用胶箔盖住，在黑暗中室温下孵育20分钟。之后，取出铝箔，将含有两毫升EDTA的150微升PBS加入每井进行清洗。再次用粘合箔盖住板。在300倍G下离心，在室温下离心5分钟。然后取出铝箔，快速、有力地将盘子倒置在水槽或一次性纸巾上，然后丢弃上一杯。将含有2毫摩尔EDTA的200微升PBS中的细胞重新悬浮，将细胞悬浮液从每一个井转移到一个单独的圆形底部管中。启动流动细胞仪，让它预热。在采集软件中，创建新的实验，并设置新样本并标记新样本。设置适当的荧光团的参数和探测器，如文本协议中所述。在此软件中，显示文本协议中概述的适当点图。只使用细胞样本，获取一些细胞，并在白细胞周围设置门。基于白细胞门，CD45正细胞的门，从SSCCD45图中与康尼迪亚分离。在点图中，CD14、CD66b、门中性粒细胞和单核细胞分别。在此之后，从仅细胞样本更改为未标记的 Conidia。在点图中，反 FITC、APC FITC 在用于反 FITC 的设置象限中显示嗜中性粒细胞，以及 FITC 信号。打开统计视图并调整象限，允许象限 Q1、Q2 和 Q4 中最多 1% 的单元格。对单核门重复此过程。在白细胞门中，记录所有至少有2万个事件的样品。从统计学中可以读取由人类嗜中性粒细胞标记的 FITC 标记的 Conidia 的噬菌体。本研究中，采用快速流动细胞测量方法测量阿斯佩吉卢斯熏蒸器与大量原生人类白细胞的相互作用。使用适当的抗体和此处显示的浇注策略，FSC 和 SSC 特征对人白细胞进行一般浇注，随后由泛白细胞标记 CD45 分离自由 Conidia 中的白细胞。Conidia可以在流式细胞测量时达到几乎细胞大小，特别是在使用肿胀的康迪迪亚时，或长时间的孵育时间，因此可以买我们分析。由于人类原发性单核细胞和中性粒细胞对 Conidia 的影响不同，因此该协议允许基于与成熟的细胞系标记进行染色、单核细胞的 CD14 和用于嗜中性粒细胞的 CD66b 来单独分析这些细胞群体。人类原发性噬细胞内化的柯尼迪亚的百分比在献血者中变化很大，但也取决于实验因素，如孵化时间和康尼迪亚的膨胀状态。孢子内化可以在0.5小时的共育后检测到，并随着时间的推移而增加。即使潜伏时间很短，预膨胀的康尼迪亚也比休息孢子更容易。如果康尼迪亚用甲醛固定，与本地康迪迪亚相比，噬菌体病会减少。按照这个程序，伊丽莎可以收集和分析共同孵化免疫细胞和康尼迪亚的超级细胞，以检查相互作用是否引起免疫细胞释放细胞因子。请记住，人类血液可能传播传染病。这项工作需要接种乙型肝炎疫苗，使用生物安全柜，以及戴手套在实验室外套。

Research

JoVE Journal

Immunology and Infection

Measuring Phagocytosis of Aspergillus fumigatus Conidia by Human Leukocytes using Flow Cytometry

Measuring Calpain Activity in Fixed and Living Cells by Flow Cytometry

在固定和活细胞，流式细胞仪测量钙蛋白酶活性

Calpains are ubiquitous intracellular, calcium-sensitive, neutral cysteine proteases 1. Calpains play crucial roles in many physiological processes, ...

科研

免疫与感染

Human In Vitro Suppression as Screening Tool for the Recognition of an Early State of Immune Imbalance

Human In Vitro Suppression as Screening Tool for the Recognition of an Early State of Immune Imbalance

人类<em在体外承认免疫失衡的早期国家禁止作为筛检工具

Regulatory T cells (Tregs) are critical mediators of immune tolerance to self-antigens. In addition, they are crucial regulators of the immune response ...

Measuring Attachment and Internalization of Influenza A Virus in A549 Cells by Flow Cytometry

通过流式细胞仪测量附件和流感的内在病毒在A549细胞

Attachment to target cells followed by internalization are the very first steps of the life cycle of influenza A virus (IAV). We provide here a detailed ...

Measuring Granulocyte and Monocyte Phagocytosis and Oxidative Burst Activity in Human Blood

人体血液中测粒及单核细胞吞噬功能和呼吸爆发活动

The granulocyte and monocyte phagocytosis and oxidative burst (OB) activity assay can be used to study the innate immune system. This manuscript provides ...

Live Imaging of Antifungal Activity by Human Primary Neutrophils and Monocytes in Response to A. fumigatus

Live Imaging of Antifungal Activity by Human Primary Neutrophils and Monocytes in Response to A. fumigatus

为了应对抑菌活性的实时成像人力初级中性粒细胞和单核细胞<em&gt;一种。曲霉</em

Aspergillus fumigatus is an opportunistic fungal pathogen causing invasive infections in immunocompromised hosts with a high case-fatality rate. Research ...

Isolation and In Vitro Culture of Murine and Human Alveolar Macrophages

Isolation and In Vitro Culture of Murine and Human Alveolar Macrophages

小鼠与人肺泡巨噬细胞的分离和体外培养

Alveolar macrophages are terminally differentiated, lung-resident macrophages of prenatal origin. Alveolar macrophages are unique in their long life and ...

Characterization of Human Monocyte Subsets by Whole Blood Flow Cytometry Analysis

全血流式细胞术分析人单核细胞亚群的表征

Monocytes are key contributors in various inflammatory disorders and alterations to these cells, including their subset proportions and functions, can ...

Isolation of Leukocytes from Human Breast Milk for Use in an Antibody-dependent Cellular Phagocytosis Assay of HIV Targets

从人类母乳中分离白细胞，用于HIV靶点抗体依赖性细胞噬菌体测定

Even in the absence of antiretroviral drugs, only ~15% of infants breastfed by HIV-infected mothers become infected, suggesting a strong protective effect ...

Measuring Erythrocyte Complement Receptor 1 Using Flow Cytometry

使用流细胞测定测量红细胞补充受体1

CR1 (CD35, Complement Receptor type 1 for C3b/C4b) is a high molecular weight membrane glycoprotein of about 200 kDa that controls complement activation, ...