Name: flow cytometry-based assay for the monitoring of nk cell functions
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Authors were supported by the German Cancer Aid (Max Eder Nachwuchsgruppe, Deutsche Krebshilfe; EU), the LOEWE Center for Cell and Gene Therapy Frankfurt (EU, ST) funded by the Hessian Ministry of Higher Education, Research and the Arts, Germany (III L 4- 518/17.004) and by the "Alfred- und Angelika Gutermuth-Stiftung", Frankfurt, Germany (EU). BJ was funded by a Mildred Scheel postdoctoral scholarship from the Dr. Mildred Scheel Foundation for Cancer Research. ST was founded by a GO-IN postdoctoral fellowship (PCOFUND-GA-2011-291776). The authors thank Becton Dickinson (BD) for providing the FACSCanto II and Canto10c Flow Cytometry Analyzers used in this study.

Bu çalışma Frankfurt Üniversitesi yerel etik kurul önerileri doğrultusunda gerçekleştirilmiştir. K562 hücrelerinin 1. Kültürleme <ol><li> R10 ortam Kültür K562 hücreleri, 37 ° C&#39;de ve% 5 CO bir hücre kültürü şişesi içinde, ml başına 0.5-1 x 10 6 hücre yoğunluğunda (glutamin ortamı,% 1 penisilin / streptomisin,% 10 fetal dana serumu ile RPMI-1640) 2. </li><li> Hasat K562 Hücreler Yeni deney başlamadan önce 24 saat. <ol><li> inkübatör K562 hücreleri i?...

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	<li>Watzl, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=How+to+trigger+a+killer:+modulation+of+natural+killer+cell+reactivity+on+many+levels.">How to trigger a killer: modulation of natural killer cell reactivity on many levels.</a> Adv Immunol. 124, 137-170 (2014).</li><li>Khaznadar, 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=Defective+NK+Cells+in+Acute+Myeloid+Leukemia+Patients+at+Diagnosis+Are+Associated+with+Blast+Transcriptional+Signatures+of+Immune+Evasion.">Defective NK Cells in Acute Myeloid Leukemia Patients at Diagnosis Are Associated with Blast Transcriptional Signatures of Immune Evasion.</a> J Immunol. 195 (6), 2580-2590 (2015).</li><li>Pical-Izard, 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=Reconstitution+of+natural+killer+cells+in+HLA-matched+HSCT+after+reduced-intensity+conditioning:+impact+on+clinical+outcome.">Reconstitution of natural killer cells in HLA-matched HSCT after reduced-intensity conditioning: impact on clinical outcome.</a> Biol Blood Marrow Transplant. 21 (3), 429-439 (2015).</li><li>Ahlenstiel, 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=Early+changes+in+natural+killer+cell+function+indicate+virologic+response+to+interferon+therapy+for+hepatitis+C.">Early changes in natural killer cell function indicate virologic response to interferon therapy for hepatitis C.</a> Gastroenterology. 141 (4), 1231-1239 (2011).</li><li>Porrata, L. 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tarif edilen yöntem sağlıklı vericilerden veya hastanın tam kan örnekleri NK hücre fonksiyonlarını incelemek için kolay, hızlı ve güvenilir bir yaklaşımdır. Bu yöntem, kaçınarak doğrudan tam kan NK hücrelerinin saflaştırılması için büyük bir avantaj sunmaktadır zaman alıcı bir çok saflaştırma yöntemleri 15 için zorunlu olan yoğunluk gradyan santrifüjleme. Ayrıca, pediatrik ve / veya bağışıklık eksikliği olan hastalarda örnekler için uygun bir alternatif yapar &qu...

doğuştan gelen bağışıklık sistemi doğal katil bir parçası olarak (NK) hücreleri virüs bulaşmış ya da malign hücre transformasyonu karşı ilk savunma hattı katkıda bulunur. inhibe edici ve aktive edici reseptör bir sistem T hücrelerinin aksine önceki antijen astarsız sağlıklı ve dönüştürülmüş hücreler arasında ayrım sağlar. Hedef hücre karşılaşma üzerine NK hücreleri hedeflerini öldürmek için bağışıklık sinaps içine sitotoksik granül (örneğin, perforin, granzymes) içeriğini bırakın. Hedef hücre üzerinde (MIP-1β, örneğin, makrofaj enflamatuar protein 1β) ve kemokinleri:;: Ayrıca, NK hücreleri, farklı sitokin türde (TNF-α, tümör nekroz faktörü-α, IFN-γ, örneğin, interferon-γ) üreten ve salgılayan etkileşimi veya sitokin stimülasyonu 1. Böyle sitotoksisite, kemokin ve sitokin üretimi yeterli NK hücre fonksiyonları farklı dis kaderi üzerinde önemli bir etkiye sahipkolaylaştırır. Onlar azaltılmış IFN-γ üretimi ve NK hücre reseptörlerini 2 aktive azaltılmış ifade oluşan tanı anındaki bir kusurlu NK hücre profili sergileyen eğer Lösemi hastaları nüks oranları artış göstermektedir. Hedef hücre etkileşimi üzerine sitokin üretimi de dahil olmak üzere, NK hücre sayıları ve fonksiyonu bir erken toparlanma allojenik kök hücre transplantasyonu 3 alan hastalarda azalmış nüks ve sağkalım oranı ile ilişkilidir. Ayrıca, hepatit C virüsü ile enfekte hastalarda interferon tedavisinin başlaması ile periferik NK hücrelerinin degranülasyonu kapasitesi olmayan yanıt 4 daha erken yanıt güçlüdür. NK hücre sayıları (&gt; 80 / ul) lenfoma veya multipl miyelom muzdarip hastalarda otolog kök hücre nakli (autoSCT) gün sonra 15 geliştirilmiş bir ilerlemesinde ve genel sağkalım 5 için öngörü vardır. melanom hastalarında T-hücresi ekspresyonu İmmünoglobulin ve müsin-alan-containing molekül-3 (TİM-3), NK hücreleri üzerinde bağışıklık-düzenleyici protein, bir hastalık aşaması ve prognoz 6 ile bağlantılıdır. Bilim adamları son yıllarda boyunca NK hücre fonksiyonlarını takip var. Önceden astarsız tümör hücrelerine karşı NK hücre sitotoksisite başlangıç analizi, 51Cr-açığa çıkarma deneyinde 7 ile ele alındı. Daha yakın zamanda, bilim adamları, genişletilmiş NK hücrelerinin 8 sitotoksisitesini değerlendirmek için radyoaktif olmayan bir yöntem geliştirdi. Sitokin ve kemokin üretimi sıklıkla enzim bağlantılı immünosorbent deneyi (ELISA) tekniği 9,10 kullanılarak değerlendirilmiştir. Son yıllarda bu yöntemler flow sitometri tabanlı analizleri ile tamamlanmıştır. Geleneksel yüzey boyama protokolleri ile birlikte protein taşıma inhibitörleri (örneğin, Brefeldin A ve monensin) ve hücre permeabilization yöntemlerinin kullanımı, farklı spesifik lenf kemokin ve sitokin üretimini incelemek için bilim adamları sağladıocyte alt-kümeleri (örneğin, T, B ya da NK hücreleri) 11. Ayrıca, farklı akış sitometri bazlı deneyler T ve NK hücre sitotoksisitesini izlemek için geliştirilmiştir. 2004 Alter ve ark., Sitotoksik granüller 12 degranülasyonunun için bir markör olarak hedef hücre karşılaşıldığında NK hücreleri üzerinde lizozom ilişkili protein CD 107a (LAMP1) yüzey ekspresyonunu tarif. Farklı fluorochromes ve çok kanallı akış sitometrelerinde geniş bir yelpazede günümüzde mevcut olduğundan, aynı anda farklı NK hücre alt gruplarında çeşitli NK hücre fonksiyonlarını (sitotoksisite, sitokin ve kemokin üretimi) izlenmesi mümkün hale gelmiştir. Bu biyopsi veya lökopeni mustarip hastaların kan örneklerinde, örneğin Örnek büyüklüğü sınırlı olduğu durumlarda özellikle önemli hale gelir. Global NK hücresi fonksiyonlarını test etmek için, farklı akış sitometri bazlı deneyler etkin birleştirilebilir. Theorell ve ark. İşitme cihazı uyarılır NK hücrelerilthy tümör hücre çizgisi K562 ile bağışçıların ve 13 flow sitometri aracılığı ile NK hücre degranülasyonu, iç-dış sinyal ve kemokin üretimini analiz. Son zamanlarda NK hücre alt grupları, autoSCT sırasında fenotipleri ve tümör hastalarında fonksiyonları sitometri tabanlı deneyler akış kullanılarak analiz edildi. Bu NK hücreleri degranüle ve autoSCT 11 sonra çok erken zaman noktalarında tümör hücre tanıma üzerine sitokinler / kemokinleri üretmek mümkün olduğu gösterilmiştir. Burada bir protokol degranülasyonu kapasitesi, kemokin ve sitokin üretimi dahil olmak üzere tümör hücrelerinin mümkün aynı anda farklı alt NK hücre fonksiyonlarını izlemek için yapan bir akış sitometrisi bazlı analiz kullamlarak etkileşimin ardından NK hücre fonksiyonlarını değerlendirmek için tarif edilmektedir.

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			<td height="21" style="height:21px;width:216px;">RPMI 1640 + glutamine</td>
			<td style="width:135px;">Invitrogen</td>
			<td style="width:119px;">6187-044</td>
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		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">penicilin/streptomycin</td>
			<td style="width:135px;">Invitrogen</td>
			<td style="width:119px;">15140-122</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">BD Falcon Round Bottom Tube</td>
			<td style="width:135px;">BD</td>
			<td style="width:119px;">352008</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">fetal calf serum</td>
			<td style="width:135px;">Invitrogen</td>
			<td style="width:119px;">10270-106</td>
			<td style="width:167px;">heat inactivated before use</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">T-flask</td>
			<td style="width:135px;">Greiner Bio-One</td>
			<td style="width:119px;">690195</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">K562 tumor cell line</td>
			<td style="width:135px;">DSMZ GmbH</td>
			<td style="width:119px;">ACC 10</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">ammonium-chloride-potassium (ACK) lysis buffer</td>
			<td style="width:135px;">made in house</td>
			<td style="width:119px;">/</td>
			<td style="width:167px;">components are listed in the text</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Distilled water: Ampuwa Sp&#252;ll&#246;sung 1000ml Plastipur</td>
			<td style="width:135px;">Fresenius Kabi</td>
			<td style="width:119px;">1088813</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Megafuge 40R Centrifuge</td>
			<td>Heraeus</td>
			<td style="width:119px;">/</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">EDTA blood collector tubes</td>
			<td style="width:135px;">Sarstedt</td>
			<td style="width:119px;">386453</td>
			<td style="width:167px;">S-Monovette 7,5 ml, K3 EDTA</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">UltraPure 0.5M EDTA, pH 8.0</td>
			<td>Life Technologies</td>
			<td style="width:119px;">15575-020</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Hematopoietic media (XVIVO)</td>
			<td>Lonza Group Ltd</td>
			<td style="width:119px;">BE04-743Q</td>
			<td></td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:216px;">human serum</td>
			<td style="width:135px;">DRK Blutspendedienst, Frankfurt/M&#160;</td>
			<td style="width:119px;">/</td>
			<td style="width:167px;">healthy donors with blood type AB; heat inactivated before use</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Neubauer-improved counting chamber, bright line</td>
			<td style="width:135px;">Marienfeld superior/ LO-Laboroptik Ltd.</td>
			<td style="width:119px;">0640030/ 1110000</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">trypan blue solution (0,4%)</td>
			<td>Invitrogen</td>
			<td style="width:119px;">15250-061</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">3% acetic acid with methylene blue</td>
			<td style="width:135px;">Stemcell Technologies&#160;</td>
			<td style="width:119px;">07060</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Corning 96 Well Clear V-Bottom TC-treated Microplates</td>
			<td style="width:135px;">Corning&#160;</td>
			<td style="width:119px;">3894</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Falcon 96 Well Round Bottom Not Treated Microplates</td>
			<td style="width:135px;">Corning&#160;</td>
			<td style="width:119px;">351177</td>
			<td></td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;width:216px;">DPBS (Ca2+- and Mg2+-free)</td>
			<td style="width:135px;">Gibco Invitrogen</td>
			<td style="width:119px;">14190-169</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">BSA</td>
			<td style="width:135px;">Sigma Aldrich</td>
			<td style="width:119px;">A2153-100G</td>
			<td></td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;width:216px;">NaN3</td>
			<td style="width:135px;">Sigma Aldrich</td>
			<td style="width:119px;">08591-1ML-F</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">phorbol 12-myristate 13-acetate (PMA)&#160;</td>
			<td style="width:135px;">Merck</td>
			<td style="width:119px;">524400-1MG</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">ionomycin&#160;</td>
			<td style="width:135px;">PromoKine</td>
			<td style="width:119px;">PK-CA577-1566-5</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">interleukin 15 (IL-15)</td>
			<td>PeproTech</td>
			<td>200-15</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Proleukin S (IL-2)&#160;</td>
			<td style="width:135px;">Novartis Pharma</td>
			<td style="width:119px;">730523</td>
			<td></td>
		</tr>
		<tr height="126">
			<td height="126" style="height:126px;width:216px;">Golgi Stop, Protein Transport Inhibitor (containing Monensin)</td>
			<td style="width:135px;">BD Biosciences</td>
			<td style="width:119px;">554724</td>
			<td style="width:167px;">This product can be used in combination or instead of Golgi Plug. The best combination for the wished experimental setting has to be tested.</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Golgi Plug, Protein Transport Inhibitor (containing Brefeldin A)</td>
			<td style="width:135px;">BD Biosciences</td>
			<td style="width:119px;">555029</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">paraformaldehyde</td>
			<td style="width:135px;">AppliChem</td>
			<td style="width:119px;">UN2209</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">saponin</td>
			<td style="width:135px;">Sigma Aldrich</td>
			<td style="width:119px;">47036</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">flow cytometer: Canto10C</td>
			<td style="width:135px;">BD Biosciences</td>
			<td style="width:119px;">/</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">FlowJo</td>
			<td style="width:135px;">TreeStar Inc.</td>
			<td style="width:119px;">/</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Graph Pad</td>
			<td style="width:135px;">Graph Pad Inc.</td>
			<td style="width:119px;">/</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">MACSxpress Separator</td>
			<td style="width:135px;">Miltenyi Biotec&#160;</td>
			<td style="width:119px;">130-098-308</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">MACSxpress NK isolation kit</td>
			<td style="width:135px;">Miltenyi Biotec&#160;</td>
			<td style="width:119px;">130-098-185</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">MACSxpress Erythrocyte Depletion Kit, human</td>
			<td style="width:135px;">Miltenyi Biotec&#160;</td>
			<td style="width:119px;">130-098-196</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">MACSmix Tube Rotator&#160;</td>
			<td style="width:135px;">Miltenyi Biotec&#160;</td>
			<td style="width:119px;">130-090-753</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">anti-human CD3 APC</td>
			<td style="width:135px;">Biolegend</td>
			<td>300412</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">anti-human CD3 V450</td>
			<td style="width:135px;">BD Biosciences</td>
			<td>560366</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">anti-human CD14 PerCP</td>
			<td style="width:135px;">Miltenyi Biotec&#160;</td>
			<td>130-094-969</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">anti-human CD14 V450</td>
			<td style="width:135px;">BD Biosciences</td>
			<td>560349</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">anti-human CD16 PE</td>
			<td style="width:135px;">Biolegend</td>
			<td>302008</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">anti-human CD16 PerCP</td>
			<td style="width:135px;">Biolegend</td>
			<td>302029</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">anti-human CD19 PE-Cy7</td>
			<td style="width:135px;">Biolegend</td>
			<td>302216</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">anti-human CD19 V450</td>
			<td style="width:135px;">BD Biosciences</td>
			<td>560353</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">anti-human CD45 BV510</td>
			<td style="width:135px;">BD Biosciences</td>
			<td>563204</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">anti-human CD56 FITC</td>
			<td style="width:135px;">Biolegend</td>
			<td>345811</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">anti-human CD107a PE</td>
			<td style="width:135px;">Biolegend</td>
			<td>328608</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">anti-human IFN-&#947; AF-647</td>
			<td style="width:135px;">BD Biosciences</td>
			<td>557729</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">anti-human MIP-1&#946; APC-H7</td>
			<td style="width:135px;">BD Biosciences</td>
			<td>561280</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">DAPI</td>
			<td style="width:135px;">Biolegend</td>
			<td>422801</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Zombie Violet Fixable Viability Kit</td>
			<td>Biolegend</td>
			<td>423113</td>
			<td>fixable dead cell marker</td>
		</tr>
	</tbody>
</table>

flow cytometry-based assay for the monitoring of nk cell functions

Natural killer (NK) cells are an important part of the human tumor immune surveillance system. NK cells are able to distinguish between healthy and virus-infected or malignantly transformed cells due to a set of germline encoded inhibitory and activating receptors. Upon virus or tumor cell recognition a variety of different NK cell functions are initiated including cytotoxicity against the target cell as well as cytokine and chemokine production leading to the activation of other immune cells. It has been demonstrated that accurate NK cell functions are crucial for the treatment outcome of different virus infections and malignant diseases. Here a simple and reliable method is described to analyze different NK cell functions using a flow cytometry-based assay. NK cell functions can be evaluated not only for the whole NK cell population, but also for different NK cell subsets. This technique enables scientists to easily study NK cell functions in healthy donors or patients in order to reveal their impact on different malignancies and to further discover new therapeutic strategies.

<html> Bütün NK hücre ve üç farklı NK hücresi alt kümelerinin degranülasyonu, sitokin ve kemokin üretimini analiz etmek için Ayırıcı stratejisi, Şekil 1 &#39;de gösterilmiştir. Bir sağlıklı donörün Örnek sonuçlar Şekil 2&#39;de gösterilmektedir. Bir uyarıcı olmadan NK hücreleri de IFN-y veya MIP-1 p üretilen ve yüzeyi üzerinde CD 107a (Şekil 2A)...

Watch this Scientific Journal Video about Flow Cytometry-based Assay for the Monitoring of NK Cell Functions at JoVE.com

Flow Cytometry-based Assay for the Monitoring of NK Cell Functions

Basit ve güvenilir bir yöntem, örneğin, farklı NK hücresi alt-gruplar içinde degranülasyonu, sitokin ve kemokin üretimi gibi NK hücresi fonksiyonlarının bir dizi analiz için tarif edilmektedir.

Akış NK Hücre Fonksiyonları İzleme Testi Sitometri tabanlı

Natural killer (NK) cells are an important part of the human tumor immune surveillance system. NK cells are able to distinguish between healthy and ...

Research

JoVE Journal

Immunology and Infection

Monitoring Dendritic Cell Migration using 19F / 1H Magnetic Resonance Imaging

Monitoring Dendritic Cell Migration using 19F / 1H Magnetic Resonance Imaging

Kullanarak İzleme Dendritik Hücre Göç<sup&gt; 19</sup&gt; F /<sup&gt; 1</sup&gt; H Manyetik Rezonans Görüntüleme

Continuous  advancements in noninvasive imaging modalities such as magnetic resonance  imaging (MRI) have greatly improved our ability to study ...

Cell-based Flow Cytometry Assay to Measure Cytotoxic Activity

Cytolytic activity of CD8+ T cells is rarely evaluated. We describe here a new cell-based assay to measure the capacity of antigen-specific CD8+ T cells ...

Analyzing the Functions of Mast Cells In Vivo Using 'Mast Cell Knock-in' Mice

Analyzing the Functions of Mast Cells In Vivo Using 'Mast Cell Knock-in' Mice

' MastCell Knock-in'Fareleri Kullanarak Mast Hücrelerinin İşlevlerini Analiz Etme

Mast cells (MCs) are hematopoietic cells which reside in various tissues, and are especially abundant at sites exposed to the external environment, such ...

Isolation and Flow Cytometric Analysis of Immune Cells from the Ischemic Mouse Brain

İskemik Fare Beyin gelen Bağışıklık Hücreleri İzolasyon ve Akım Sitometrisi Analizi

Ischemic stroke initiates a robust inflammatory response that starts in the intravascular compartment and involves rapid activation of brain resident ...

Fluorescence-activated Cell Sorting for Purification of Plasmacytoid Dendritic Cells from the Mouse Bone Marrow

Fare kemik iliğinden plazmasitoid dendritik hücreleri Saflaştırılması floresans ile aktive edilen hücre sınıflandırma

Fluorescence-activated cell sorting (FACS) is a technique to purify specific cell populations based on phenotypes detected by flow cytometry. This method ...

Assessing Retinal Microglial Phagocytic Function In Vivo Using a Flow Cytometry-based Assay

Assessing Retinal Microglial Phagocytic Function In Vivo Using a Flow Cytometry-based Assay

Retina Mikroglial Fagositik Fonksiyonu değerlendirilmesi<em&gt; İn Vivo</em&gt; A Akış Sitometri bazlı analiz kullamlarak

Microglia are the tissue resident macrophages of the central nervous system (CNS) and they perform a variety of functions that support CNS homeostasis, ...

Dextran Enhances the Lentiviral Transduction Efficiency of Murine and Human Primary NK Cells

Dextran fare ve insan birincil NK hücreleri Lentiviral iletim verimliliğini artırır

The efficient transduction of specific genes into natural killer (NK) cells has been a major challenge. Successful transductions are critical to defining ...

A Flow Cytometry-Based Cytotoxicity Assay for the Assessment of Human NK Cell Activity

Akış Sitometresi tabanlı sitotoksisite tahlil insan NK hücre etkinliği değerlendirilmesi için

Within the innate immune system, effector lymphocytes known as natural killer (NK) cells play an essential role in host defense against aberrant cells, ...

A Simple Flow Cytometry Based Assay to Determine In Vitro Antibody Dependent Enhancement of Dengue Virus Using Zika Virus Convalescent Serum

A Simple Flow Cytometry Based Assay to Determine In Vitro Antibody Dependent Enhancement of Dengue Virus Using Zika Virus Convalescent Serum

Basit Akış Sitometresi Vitro antikor bağımlı Enhancement-in dang virüs Zika virüs nekahat Serum kullanarak belirlemek için tahlil dayalı

Antibody dependent enhancement of infection has been shown to play a major role in Dengue viral pathogenesis. Traditional assays that measure the capacity ...

Multicolor Flow Cytometry-based Quantification of Mitochondria and Lysosomes in T Cells

Çok renkli akış sitometresi tabanlı Quantification mitokondri ve T hücrelerde organellerin

T cells utilize different metabolic programs to match their functional needs during differentiation and proliferation. Mitochondria are crucial cellular ...