Name: infection of primary nasal epithelial cells grown at an air-liquid interface to characterize human coronavirus-host interactions
Uploaded: 2023-09-22T13:20:00
Description: Watch this Scientific Journal Video about Infection of Primary Nasal Epithelial Cells Grown at an Air-Liquid Interface to Characterize Human Coronavirus-Host Interactions at JoVE.com

この研究には、次の資金源があります:国立衛生研究所(NIH)R01AI 169537(SRWおよびN.A.C.)、NIH R01AI 140442(S.R.W.)、VA Merit Review CX001717(N.A.C.)、VA Merit Review BX005432(S.R.W.およびN.A.C.)、Penn Center for Research on Coronaviruses and other Emerging Pathogens(S.R.W.)、Laffey-McHugh Foundation(S.R.W.およびN.A.C.)、 T32 AI055400 (CJO)、T32 AI007324 (AF)。

鼻標本の使用は、ペンシルベニア大学治験審査委員会(プロトコル#800614)およびフィラデルフィアVA治験審査委員会(プロトコル#00781)によって承認されました。
1. 鼻腔ALI培養の感染
注:臨床検体の取得、および鼻腔ALI培養の増殖と分化は、この論文の範囲外です。初代鼻上皮細胞を培養するための特定の方法は、これらの培養物を利用した最?...

<ol>
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R., Regamey, 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+airway+epithelium:+Soldier+in+the+fight+against+respiratory+viruses.">The airway epithelium: Soldier in the fight against respiratory viruses.</a> Clinical Microbiology Reviews. 24 (1), 210-229 (2011).</li><li>Farzal, Z., 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=Comparative+study+of+simulated+nebulized+and+spray+particle+deposition+in+chronic+rhinosinusitis+patients.">Comparative study of simulated nebulized and spray particle deposition in chronic rhinosinusitis patients.</a> International Forum of Allergy and Rhinology. 9 (7), 746-758 (2019).</li><li>Gaeckle, N. T., Pragman, A. A., Pendleton, K. M., Baldomero, A. K., Criner, G. 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F., 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=Delayed+induction+of+type+I+and+III+interferons+mediates+nasal+epithelial+cell+permissiveness+to+SARS-CoV-2.">Delayed induction of type I and III interferons mediates nasal epithelial cell permissiveness to SARS-CoV-2.</a> Nature Communications. 12 (1), 7092 (2021).</li><li>Sungnak, W., 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=SARS-CoV-2+entry+factors+are+highly+expressed+in+nasal+epithelial+cells+together+with+innate+immune+genes.">SARS-CoV-2 entry factors are highly expressed in nasal epithelial cells together with innate immune genes.</a> Nature Medicine. 26 (5), 681-687 (2020).</li><li>Li, Y., 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=SARS-CoV-2+induces+double-stranded+RNA-mediated+innate+immune+responses+in+respiratory+epithelial-derived+cells+and+cardiomyocytes.">SARS-CoV-2 induces double-stranded RNA-mediated innate immune responses in respiratory epithelial-derived cells and cardiomyocytes.</a> Proceedings of the National Academy of Sciences of the United States of America. 118 (16), 2022643118 (2021).</li><li>Comar, C. E., 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=MERS-CoV+endoribonuclease+and+accessory+proteins+jointly+evade+host+innate+immunity+during+infection+of+lung+and+nasal+epithelial+cells.">MERS-CoV endoribonuclease and accessory proteins jointly evade host innate immunity during infection of lung and nasal epithelial cells.</a> Proceedings of the National Academy of Sciences of the United States of America. 119 (21), 2123208119 (2022).</li><li>Lee, R. J., 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=Bacterial+D-amino+acids+suppress+sinonasal+innate+immunity+through+sweet+taste+receptors+in+solitary+chemosensory+cells.">Bacterial D-amino acids suppress sinonasal innate immunity through sweet taste receptors in solitary chemosensory cells.</a> Science Signaling. 10 (495), (2017).</li><li>Brewington, J. J., 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=Brushed+nasal+epithelial+cells+are+a+surrogate+for+bronchial+epithelial+CFTR+studies.">Brushed nasal epithelial cells are a surrogate for bronchial epithelial CFTR studies.</a> JCI Insight. 3 (13), 99385 (2018).</li><li>Comer, D. M., Elborn, J. S., Ennis, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparison+of+nasal+and+bronchial+epithelial+cells+obtained+from+patients+with+COPD.">Comparison of nasal and bronchial epithelial cells obtained from patients with COPD.</a> PLoS One. 7 (3), e32924 (2012).</li><li>Vanders, R. L., Hsu, A., Gibson, P. G., Murphy, V. E., Wark, P. A. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Nasal+epithelial+cells+to+assess+in+vitro+immune+responses+to+respiratory+virus+infection+in+pregnant+women+with+asthma.">Nasal epithelial cells to assess in vitro immune responses to respiratory virus infection in pregnant women with asthma.</a> Respiratory Research. 20 (1), 259 (2019).</li><li>Lee, R. J., 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=Fungal+aflatoxins+reduce+respiratory+mucosal+ciliary+function.">Fungal aflatoxins reduce respiratory mucosal ciliary function.</a> Scientific Reports. 6, 33221 (2016).</li><li>Patel, N. N., 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=Fungal+extracts+stimulate+solitary+chemosensory+cell+expansion+in+noninvasive+fungal+rhinosinusitis.">Fungal extracts stimulate solitary chemosensory cell expansion in noninvasive fungal rhinosinusitis.</a> International Forum of Allergy and Rhinology. 9 (7), 730-737 (2019).</li><li>Baer, A., Kehn-Hall, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Viral+concentration+determination+through+plaque+assays:+Using+traditional+and+novel+overlay+systems.">Viral concentration determination through plaque assays: Using traditional and novel overlay systems.</a> Journal of Visualized Experiments. 93 (93), e52065 (2014).</li><li>Robinot, 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=SARS-CoV-2+infection+induces+the+dedifferentiation+of+multiciliated+cells+and+impairs+mucociliary+clearance.">SARS-CoV-2 infection induces the dedifferentiation of multiciliated cells and impairs mucociliary clearance.</a> Nature Communications. 12 (1), 4354 (2021).</li><li>Whitsett, 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=Airway+epithelial+differentiation+and+mucociliary+clearance.">Airway epithelial differentiation and mucociliary clearance.</a> Annals of the American Thoracic Society. 15, S143-S148 (2018).</li><li>Gao, N., Raduka, A., Rezaee, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Respiratory+syncytial+virus+disrupts+the+airway+epithelial+barrier+by+decreasing+cortactin+and+destabilizing+F-actin.">Respiratory syncytial virus disrupts the airway epithelial barrier by decreasing cortactin and destabilizing F-actin.</a> Journal of Cell Science. 135 (16), 259871 (2022).</li><li>Schmidt, H., 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=IL-13+impairs+tight+junctions+in+airway+epithelia.">IL-13 impairs tight junctions in airway epithelia.</a> International Journal of Molecular Sciences. 20 (13), 3222 (2019).</li><li>Huang, Z. Q., 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=Interleukin-13+alters+tight+junction+proteins+expression+thereby+compromising+barrier+function+and+dampens+rhinovirus+induced+immune+responses+in+nasal+epithelium.">Interleukin-13 alters tight junction proteins expression thereby compromising barrier function and dampens rhinovirus induced immune responses in nasal epithelium.</a> Frontiers in Cell and Developmental Biology. 8, 572749 (2020).</li><li>Saatian, B., 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=Interleukin-4+and+interleukin-13+cause+barrier+dysfunction+in+human+airway+epithelial+cells.">Interleukin-4 and interleukin-13 cause barrier dysfunction in human airway epithelial cells.</a> Tissue Barriers. 1 (2), e24333 (2013).</li><li>Coles, J. 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ここで詳述する方法は、患者由来の鼻上皮細胞を空気と液体の界面で増殖させ、HCoVと宿主の相互作用の研究に適用する一次上皮培養システムについて説明しています。分化されると、これらの鼻腔ALI培養物は、繊毛細胞、杯細胞、基底細胞が表される不均一な細胞集団や、繊毛と粘液分泌を頑強に鼓動する無傷の粘膜繊毛機能など、 in vivo 鼻上皮の多くの特徴を再現します。この不均...

これまでに7種類のヒトコロナウイルス(HCoV)が確認されており、さまざまな呼吸器疾患を引き起こします2。一般的または季節性のHCoV(HCoV-NL63、-229E、-OC43、および-HKU1)は、通常、上気道の病理学に関連しており、毎年一般的な風邪症例の推定10%〜30%を引き起こします。これは一般的なHCoVに関連する典型的な臨床表現型ですが、これらのウイルスは、子供、高齢者、免疫不全の個人など、リスクのある集団でより重大な下気道疾患を引き起こす可能性があります3,4。過去20年間に、重症急性呼吸器症候群(SARS)-CoV、中東呼吸器症候群(MERS)-CoV、SARS-CoV-2の3つの病原性HCoVが出現し、重大な公衆衛生上の緊急事態を引き起こしています。致死的なHCoVは、より重篤な気道病変と関連しており、MERS-CoV症例に関連する致死率>34%(2012年の出現以来、2,500例以上で894例)5,6。致死的なHCoVは、現在進行中のCOVID-19パンデミックに見られるように、無症候性感染症から致死性肺炎まで、さまざまな気道疾患も引き起こすことに注意することが重要です7。
HCoVは、他の呼吸器病原体と同様に、気道に入り、鼻上皮に生産的な感染を確立します8。下気道への広がりは、口腔/鼻腔から肺への誤嚥に関連していると考えられており、HCoVはより重大な下気道病状を引き起こします9,10,11。したがって、鼻はウイルス侵入の最初のポータルとして機能し、その強力な粘膜繊毛クリアランス機構と、下気道へのウイルスのさらなる拡散を防ぐことを目的とした独自の自然免疫メカニズムにより、感染に対する主要な障壁となります12,13。例えば、鼻の上皮細胞は、抗ウイルスインターフェロンおよびインターフェロン刺激遺伝子の基底レベルが平均よりも高いことが報告されており、鼻細胞が呼吸器系ウイルスに対する早期応答の準備ができている可能性があることを示しています14,15,16。
私たちはこれまで、患者由来の初代鼻上皮細胞を気液界面(ALI)で増殖させ、HCoV感染が始まる鼻のHCoVと宿主の相互作用をモデル化しました。経鼻ALI培養は、病原性(SARS-CoV-2およびMERS-CoV)と一般的なHCoV(HCoV-NL63およびHCoV-229E)の両方に寛容であり、A549(肺腺癌細胞株)などの従来の気道上皮細胞株に比べてさまざまな利点があります16,17。分化後、鼻腔ALI培養物は不均一な細胞集団を含み、粘膜繊毛クリアランス機構など、in vivo鼻上皮に期待される機能の多くを示す18。鼻細胞は、細胞学的ブラッシングによる鼻上皮細胞の獲得は、HBECを達成するための気管支鏡検査などの技術の使用と比較して侵襲性が大幅に低いため、下気道培養システム(ヒト気管支上皮細胞、HBEC)よりも利点もあります19,20,21。
この論文では、この鼻腔ALI培養システムを利用して、鼻上皮におけるHCoVと宿主の相互作用を特徴付ける方法について説明します。これらの手法を最近発表された研究に適用し、SARS-CoV-2、MERS-CoV、HCoV-NL63、およびHCoV-229E 1,16,17を比較しました。これらの方法と代表的な結果は、この鼻細胞モデルにおけるHCoVの研究を強調していますが、このシステムは他のHCoVや他の呼吸器病原体にも高い適応性を持っています。さらに、これらの方法は、ウイルス複製と細胞向性、ならびに感染後の細胞毒性と自然免疫誘導を調査するために、他のALI培養系により広く適用することができます。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Alexa Fluor secondary antibodies (488, 594, 647)</td>
			<td>Invitrogen</td>
			<td>Various</td>
			<td></td>
		</tr>
		<tr>
			<td>BSA (bovine serum albumin)</td>
			<td>Sigma-Aldrich</td>
			<td>A7906</td>
			<td></td>
		</tr>
		<tr>
			<td>cOmplete mini EDTA-free protease inhibitor</td>
			<td>Roche</td>
			<td>11836170001</td>
			<td></td>
		</tr>
		<tr>
			<td>Cytotoxicity detection kit</td>
			<td>Roche</td>
			<td>11644793001</td>
			<td></td>
		</tr>
		<tr>
			<td>DMEM (Dulbecco&#39;s Modified Eagle Media)</td>
			<td>Gibco</td>
			<td>11965-084</td>
			<td></td>
		</tr>
		<tr>
			<td>DPBS (Dulbecco&#39;s Phosphate Buffered Saline)</td>
			<td>Gibco</td>
			<td>14190136</td>
			<td></td>
		</tr>
		<tr>
			<td>DPBS + calcium + magnesium</td>
			<td>Gibco</td>
			<td>14040-117</td>
			<td></td>
		</tr>
		<tr>
			<td>Endohm-6G measurement chamber</td>
			<td>World Precision Instruments</td>
			<td>ENDOHM-6G</td>
			<td></td>
		</tr>
		<tr>
			<td>Epithelial cell adhesion marker (EpCAM; CD326)</td>
			<td>eBiosciences</td>
			<td>14-9326-82</td>
			<td></td>
		</tr>
		<tr>
			<td>Epithelial Volt/Ohm (TEER) Meter (EVOM)</td>
			<td>World Precision Instruments</td>
			<td>300523</td>
			<td></td>
		</tr>
		<tr>
			<td>FBS (Fetal Bovine Serum)</td>
			<td>HyClone</td>
			<td>SH30071.03</td>
			<td></td>
		</tr>
		<tr>
			<td>FV10-ASW software for imaging</td>
			<td>Olympus</td>
			<td>Version 4.02</td>
			<td></td>
		</tr>
		<tr>
			<td>HCoV-NL63 (Human coronavirus, NL63)</td>
			<td>BEI Resources</td>
			<td>NR-470</td>
			<td></td>
		</tr>
		<tr>
			<td>HCoV-NL63 nucleocapsid antibody</td>
			<td>Sino Biological</td>
			<td>40641-V07E</td>
			<td></td>
		</tr>
		<tr>
			<td>Hoescht stain</td>
			<td>Thermo Fisher</td>
			<td>H3570</td>
			<td></td>
		</tr>
		<tr>
			<td>Laemmli sample buffer (4x)</td>
			<td>BIO-RAD</td>
			<td>1610747</td>
			<td></td>
		</tr>
		<tr>
			<td>LLC-MK2 cells</td>
			<td>ATCC</td>
			<td>CCL-7</td>
			<td>To titrate HCoV-NL63</td>
		</tr>
		<tr>
			<td>MERS-CoV (Human coronavirus, Middle East Respiratory Syndrome Coronavirus (MERS-CoV), EMC/2012)</td>
			<td>BEI Resources&#160;</td>
			<td>NR-44260</td>
			<td></td>
		</tr>
		<tr>
			<td>MERS-CoV nucleocapsid antibody</td>
			<td>Sino Biological</td>
			<td>40068-MM10</td>
			<td></td>
		</tr>
		<tr>
			<td>MUC5AC antibody</td>
			<td>Sigma-Aldrich</td>
			<td>AMAB91539</td>
			<td></td>
		</tr>
		<tr>
			<td>Olympus Fluoview confocal microscope</td>
			<td>Olympus</td>
			<td>FV1000</td>
			<td></td>
		</tr>
		<tr>
			<td>Phalloidin-iFluor 647 stain</td>
			<td>Abcam</td>
			<td>ab176759</td>
			<td></td>
		</tr>
		<tr>
			<td>PhosStop easy pack (phosphatase inhibitors)&#160;</td>
			<td>Roche</td>
			<td>PHOSS-RO</td>
			<td></td>
		</tr>
		<tr>
			<td>Plate reader&#160;</td>
			<td>Perkin Elmer</td>
			<td>HH34000000</td>
			<td>Any plate reader or ELISA reader is sufficient; must be able to read absorbance at 492 nm</td>
		</tr>
		<tr>
			<td>RIPA buffer (50 mM Tris pH 8; 150 mM NaCl; 0.5% deoxycholate; 0.1% SDS; 1% NP40)</td>
			<td>Thermo Fisher</td>
			<td>89990</td>
			<td>Can prep in-house or purchase</td>
		</tr>
		<tr>
			<td>RNeasy Plus Kit</td>
			<td>Qiagen</td>
			<td>74134</td>
			<td></td>
		</tr>
		<tr>
			<td>SARS-CoV-2 (SARS-Related Coronavirus 2, Isolate USA-WA1/2020)</td>
			<td>BEI Resources</td>
			<td>NR-52281</td>
			<td></td>
		</tr>
		<tr>
			<td>SARS-CoV-2 nucleocapsid antibody</td>
			<td>Genetex</td>
			<td>GTX135357</td>
			<td></td>
		</tr>
		<tr>
			<td>Triton-X 100</td>
			<td>Fisher Scientific</td>
			<td>BP151100</td>
			<td></td>
		</tr>
		<tr>
			<td>Type IV &#946;- tubulin antibody</td>
			<td>Abcam</td>
			<td>ab11315</td>
			<td></td>
		</tr>
		<tr>
			<td>VeroCCL81 cells</td>
			<td>ATCC</td>
			<td>CCL-81</td>
			<td>To titrate MERS-CoV</td>
		</tr>
		<tr>
			<td>VeroE6 cells</td>
			<td>ATCC</td>
			<td>CRL-1586</td>
			<td>To titrate SARS-CoV-2</td>
		</tr>
	</tbody>
</table>

infection of primary nasal epithelial cells grown at an air-liquid interface to characterize human coronavirus-host interactions

SARS-CoV(2002年)、MERS-CoV(2012年)、SARS-CoV-2(2019年)の3つの高病原性ヒトコロナウイルス(HCoV)が出現し、過去20年間に重大な公衆衛生上の危機を引き起こしました。毎年、風邪の症例のかなりの部分(HCoV-NL63、-229E、-OC43、-HKU1)がさらに4つのHCoVを引き起こしており、生理学的に関連するシステムでこれらのウイルスを研究することの重要性が強調されています。HCoVは気道に侵入し、すべての呼吸器病原体が遭遇する主要な部位である鼻上皮に感染を確立します。私たちは、患者由来の鼻サンプルを気液界面(ALI)で増殖させる一次鼻上皮培養システムを使用して、この重要なセンチネル部位での宿主と病原体の相互作用を研究しています。これらの培養物は、存在する細胞の種類、繊毛機能、粘液産生など、 in vivo 気道の多くの特徴を再現しています。HCoV感染後の経鼻ALI培養におけるウイルス複製、宿主細胞の向性、ウイルス誘発性細胞傷害性、および自然免疫誘導を特徴付ける方法を、致死性と季節性HCoVを比較した最近の研究を例に説明します1。鼻腔内の宿主と病原体の相互作用に関する理解が深まれば、HCoVやその他の呼吸器系ウイルスに対する抗ウイルス治療薬の新たな標的となる可能性があります。

代表的な数値は、Otter et al.1の原稿に見られるデータから部分的に改作されています。4人または6人のドナー由来の経鼻ALI培養物を、上記のプロトコルに従って4つのHCoV(SARS-CoV-2、MERS-CoV、HCoV-NL63、およびHCoV-229E)のいずれかに感染させ、各ウイルスの平均頂端排出ウイルス力価を 図1Aに示します。これら4つのHCoVはすべて経鼻ALI培養で生産的に複製されま?...

Watch this Scientific Journal Video about Infection of Primary Nasal Epithelial Cells Grown at an Air-Liquid Interface to Characterize Human Coronavirus-Host Interactions at JoVE.com

Infection of Primary Nasal Epithelial Cells Grown at an Air-Liquid Interface to Characterize Human Coronavirus-Host Interactions

鼻上皮は、すべての呼吸器病原体が遭遇する主要なバリア部位です。ここでは、空気液界面(ALI)培養物として増殖させた初代鼻上皮細胞を用いて、生理学的に関連性の高いシステムにおけるヒトコロナウイルスと宿主の相互作用を特徴付ける方法について概説します。

ヒトコロナウイルスと宿主の相互作用を特徴づけるための空気-液体界面で増殖した初代鼻上皮細胞の感染

Three highly pathogenic human coronaviruses (HCoVs) - SARS-CoV (2002), MERS-CoV (2012), and SARS-CoV-2 (2019) - have emerged and caused significant public ...

Research

JoVE Journal

Immunology and Infection

Co-culture Models of Pseudomonas aeruginosa Biofilms Grown on Live Human Airway Cells

Co-culture Models of Pseudomonas aeruginosa Biofilms Grown on Live Human Airway Cells

共培養モデルの緑膿菌</emライブヒト気道細胞上に成長した>バイオフィルム

Bacterial biofilms have been associated with a number of different human diseases, but biofilm development has generally been studied on non-living ...

Application of a Mouse Ligated Peyer&#x2019;s Patch Intestinal Loop Assay to Evaluate Bacterial Uptake by M cells

Application of a Mouse Ligated PeyerÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢s Patch Intestinal Loop Assay to Evaluate Bacterial Uptake by M cells

M細胞による細菌の取り込みを評価するパイエル板腸管ループアッセイを連結したマウスのアプリケーション

The inside of our gut is inhabited with enormous number of commensal bacteria. The mucosal surface of the gastrointestinal tract is continuously exposed ...

Following Cell-fate in E. coli After Infection by Phage Lambda

Following Cell-fate in E. coli After Infection by Phage Lambda

細胞運命を以下に<em&gt; E大腸菌の</emラムダファージによる感染後&gt;

The system comprising bacteriophage (phage) lambda and the bacterium E. coli has long served as a paradigm for cell-fate determination1,2. Following the ...

An In vitro Co-infection Model to Study Plasmodium falciparum-HIV-1 Interactions in Human Primary Monocyte-derived Immune Cells

An In vitro Co-infection Model to Study Plasmodium falciparum-HIV-1 Interactions in Human Primary Monocyte-derived Immune Cells

不定冠詞<em&gt;のin vitro</em研究する&gt;共同感染モデル<em&gt;熱帯熱マラリア原虫（Plasmodium falciparum）</emヒトの第一次単球由来の免疫細胞に&gt;-HIV-1の相互作用

Plasmodium falciparum, the causative agent  of the deadliest form of malaria, and human immunodeficiency virus type-1 (HIV-1) are among the most important ...

Flow Cytometric Isolation of Primary Murine Type II Alveolar Epithelial Cells for Functional and Molecular Studies

機能と分子生物学的研究のためのプライマリマウスII型肺胞上皮細胞のフローサイトメトリ分離

Throughout  the last years, the contribution of alveolar type II epithelial cells (AECII)  to various aspects of immune regulation in the lung has been ...

Isolation of Myeloid Dendritic Cells and Epithelial Cells from Human Thymus

ヒト胸腺からの骨髄樹状細胞と上皮細胞の単離

In this protocol we provide a method to isolate dendritic cells (DC) and epithelial cells (TEC) from the human thymus. DC and TEC are the major antigen ...

Biomimetic Materials to Characterize Bacteria-host Interactions

細菌 - 宿主相互作用を特徴づけるために、バイオミメティック材料

Bacterial attachment to host cells is one of the earliest events during bacterial colonization of host tissues and thus a key step during infection. The ...

Human Placental and Decidual Organ Cultures to Study Infections at the Maternal-fetal Interface

母体・胎児のインターフェイスでの感染症を研究するためにヒト胎盤および脱落膜器官培養

The placenta shows a large degree of interspecies anatomic variability. To best understand biology and pathophysiology of the human placenta, it is ...

Assessment of the Synaptic Interface of Primary Human T Cells from Peripheral Blood and Lymphoid Tissue

末梢血およびリンパ組織から主な人間の T 細胞のシナプスのインタ フェースの評価

The current understanding of the dynamics and structural features of T-cell synaptic interfaces has been largely determined through the use of ...

High-Efficiency Generation of Antigen-Specific Primary Mouse Cytotoxic T Cells for Functional Testing in an Autoimmune Diabetes Model

自己免疫性糖尿病モデルにおける機能試験用抗原特異的一次マウス細胞傷害性T細胞の高効率生成

Type 1 Diabetes (T1D) is characterized by islet-specific autoimmunity leading to beta cell destruction and absolute loss of insulin production. In the ...