Yazarlar deneyleri ve veri analizi sırasında destek için Dr. Julia Mannheim Walter Ehrlichmann Ramona Stumm, Funda Cay Daniel Bukala, Maren Harant yanı sıra Natalie Altmeyer teşekkür ederim. Bu çalışma Werner Siemens-Vakfı, SFB685 aracılığıyla DFG (proje B6) ve fal (2309-0-0) tarafından desteklenmiştir.

Güvenlik önlemleri: radyoaktivite kullanırken, 2-inç kalınlığında kurşun tuğla arkasında 64 Cu depolamak ve aktivitesini taşıyan gemiler için ilgili ekranlama kullanın. dolaylı dolaysız el temasını önlemek ve radyoaktif madde maruziyeti en aza indirmek için korumasız kaynaklar işlemek için uygun araçları kullanın. Her zaman radyasyon dozimetre izleme rozetleri ve kişisel koruma ekipmanları giymek ve kendini kontrol ve kirlenme için çalışma alanı hemen ele almak. önce radyoak...

<ol>
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Bu protokol, stabil bir şekilde PET in vivo izlenmesi için hücreler radyolabel güvenilir ve kolay bir yöntem sunulur. 64Cu-DOTA-KJ1-26-mAb ve homing ile de radyo-etiketlenebilir, cova-TH1 hücreleri, izole edilmiş ve donör farelerden in vitro olarak genişletilebilir, bu yöntem olabilir kullanan bir Cova sunum siteleri gibi pulmoner ve perithymic LN için takip edilmiştir akut solunum yolu DTHR cova kaynaklı. çelatör mAb modifikasyonu hızlı ve verim...

Invazif olmayan hücre takibi, in vivo olarak, hücre fonksiyonu, göç ve homing izlemek için çok yönlü bir araçtır. Son hücre takibi çalışmaları rejeneratif tıpta, kanser, 3, 4 karşı adoptif hücre tedavileri enflamasyonun veya T lenfositleri otolog periferik beyaz kan hücreleri bağlamında mesenkimal 1, 2 ya da kemik iliği kökenli kök hücreleri 3 üzerine odaklanmıştır. eylem siteleri ve hücre tabanlı tedaviler yatan biyolojik ilkelerin aydınlatılması muazzam öneme sahiptir. CD8 + sitotoksik T lenfositleri, genetik olarak yaygın altın standart olarak kabul edilmiştir kimerik antijen reseptörü (CAR), T hücreleri ya da tümör infiltre edici lenfositler (TILler) tasarlanmıştır. Bununla birlikte, tümör bağlantılı antijen-spesifik Th1 hücrelerinin &lt;etkili bir tedavi seçeneği 4, olduğu kanıtlanmıştır/ sup&gt; 5, 6, 7. Anahtar iltihaplanmada oyuncuların organa özgü otoimmün bir hastalık (örneğin, romatoid artrit veya bronşiyal astım) ve kanser immünoterapisi yüksek ilgi hücreleri gibi, TH1 hücre zamansal dağılımı ve yerleşme kalıpları karakterize etmek için önemlidir. PET ile invaziv olmayan in vivo görüntüleme in vivo hominginde, hücre göçü modellerini incelemek için bir nicel, çok hassas bir yöntem 8. sunulur, ve iltihap, alerji, enfeksiyon ya da tümör rejeksiyon 9, 10, 11 sırasında T hücresi eylem ve tepkilerinin siteleri. Klinik olarak, In-oksin 111 2-deoksi-2- (18 ise, iltihap ve enfeksiyon 12 ayrımı için lökosit sintigrafisi için kullanılırF)-floro-D-glükoz (18F-FDG) genellikle PET 3, 13 ile hücre izleme çalışmaları için kullanılır. Bu PET tracer önemli bir dezavantajı, ancak, 109.7 dk&#39;da radyonüklid 18F ve daha sonraki zaman noktalarında görüntüleme adoptif hücre transferi sonrası engelleyen düşük hücre içi stabilite kısa yarılanma ömrü. Hücrelerde kararsız olmasına rağmen, 64Cu-PTSM sık sık kullanılan PET ile in vivo hücre takibi çalışmalarında uzun vadede için, spesifik olmayan bir hücre 14, T hücre canlılığı üzerindeki minimize zararlı etkiler ile 15 etiket ve 16 işlev görmektedir. Bu protokol, ayrıca bir T hücre reseptörü (TCR) e özgü radyo-etiketli mAb kullanılarak hücre canlılığı ve işlevi üzerinde dezavantajlı etkileri azaltmak için bir yöntemi tarif etmektedir. İlk olarak, radyoizotop 64 Cu, inci ile mAb KJ1-26 konjüge üretimiE şelatör DOTA, ve daha sonra 64 Cu, radyo-etiketleme gösterilmiştir. İkinci bir aşamada, izolasyon ve DO11.10 donör farelerin cova-TH1 hücrelerinin genişlemesi ve 64Cu yüklü DOTA-konjuge mAb KJ1-26 (64Cu-DOTA-KJ1-26) ile radyo işaretli ayrıntılı olarak tarif edilmiştir. Uptake&#39;leri ve bir doz kalibratörle radyoaktivite efflux sırasıyla y-sayımı ile değerlendirme, hem de IFN-y ELISA ile sunulmuştur tripan mavi dışlama ve işlevselliği ile hücre canlılığı üzerindeki 64Cu-radyoaktif etkilerinin değerlendirilmesi . In vivo hücre takibi yayılmayan için adoptif hücre transferi sonrası PET / CT tarafından cova kaynaklı akut solunum yolu DTHR ve görüntü elde olan bir fare modeli ortaya çıkarma açıklanmaktadır. Ayrıca, bu markalama yaklaşım, farklı hastalık modellerinde farklı TCRler veya membrana bağlı reseptörlere veya ekspresyon mar ile ilgili genel hücreleri ile fare T hücreleri aktarılabilirkers sürekli zar 17 mekik yatan.

<table border="0" cellpadding="0" cellspacing="0" width="966">
	<tbody>
		<tr height="24">
			<td height="24" style="height:24px;width:292px;">HCl, Suprapur</td>
			<td style="width:227px;">Merck, Darmstadt, Germany</td>
			<td style="width:259px;">1.00318</td>
			<td style="width:189px;">64Cu production</td>
		</tr>
		<tr height="24">
			<td height="24" style="height:24px;width:292px;">Methanol, Suprapur</td>
			<td style="width:227px;">Merck, Darmstadt, Germany</td>
			<td>1.06007</td>
			<td>64Cu production</td>
		</tr>
		<tr height="24">
			<td height="24" style="height:24px;width:292px;">Isopropanol, Suprapur</td>
			<td style="width:227px;">Merck, Darmstadt, Germany</td>
			<td>1.0104</td>
			<td>64Cu production</td>
		</tr>
		<tr height="24">
			<td height="24" style="height:24px;width:292px;">Pt/Ir (90/10) plate</td>
			<td style="width:227px;">&#214;gussa</td>
			<td style="width:259px;">Custom made</td>
			<td>64Cu production</td>
		</tr>
		<tr height="24">
			<td height="24" style="height:24px;width:292px;">PEEK chamber</td>
			<td style="width:227px;">&#214;gussa</td>
			<td style="width:259px;">Custom made</td>
			<td>64Cu production</td>
		</tr>
		<tr height="24">
			<td height="24" style="height:24px;width:292px;">64Ni</td>
			<td style="width:227px;">Chemotrade</td>
			<td style="width:259px;"></td>
			<td>64Cu production</td>
		</tr>
		<tr height="24">
			<td height="24" style="height:24px;width:292px;">Polygram SIL G/UV 254 plate</td>
			<td style="width:227px;">Macherey-Nagel</td>
			<td style="width:259px;">805021</td>
			<td>64Cu production</td>
		</tr>
		<tr height="24">
			<td height="24" style="height:24px;width:292px;">Ion exchange column</td>
			<td style="width:227px;">BioRad</td>
			<td style="width:259px;">AG1-X8</td>
			<td>64Cu production</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:292px;">Solid state target system for PETtrace</td>
			<td style="width:227px;">WKL</td>
			<td style="width:259px;">costum made</td>
			<td>64Cu production</td>
		</tr>
		<tr height="24">
			<td height="24" style="height:24px;width:292px;">64Cu work-up module</td>
			<td style="width:227px;">WKL</td>
			<td style="width:259px;">costum made</td>
			<td>64Cu production</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:292px;">Dose calibrator</td>
			<td style="width:227px;">Capintec</td>
			<td style="width:259px;">CRC-25R</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:292px;">PETtrace cyclotron</td>
			<td style="width:227px;">General Electric Medical Systems</td>
			<td style="width:259px;"></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:292px;">DOTA-NHS</td>
			<td style="width:227px;">Macrocyclics</td>
			<td style="width:259px;">B-280</td>
			<td>DOTA-conjugation</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:292px;">Anti-cOVA-TCR antibody (KJ1-26)</td>
			<td style="width:227px;"></td>
			<td>Isolated from hybridoma cell culture</td>
			<td>DOTA-conjugation</td>
		</tr>
		<tr height="26">
			<td height="26" style="height:26px;width:292px;">Na2HPO4</td>
			<td style="width:227px;">Sigma-Aldrich</td>
			<td style="width:259px;">71633</td>
			<td>DOTA-conjugation</td>
		</tr>
		<tr height="24">
			<td height="24" style="height:24px;width:292px;">H+ Chelex 100</td>
			<td style="width:227px;">Sigma-Aldrich</td>
			<td style="width:259px;">C7901</td>
			<td>DOTA-conjugation</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:292px;">Amicon Ultra-15 filter unit</td>
			<td style="width:227px;">Merck Millipore</td>
			<td style="width:259px;">UFC910008</td>
			<td>DOTA-conjugation</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:292px;">Rotipuran ultrapure water</td>
			<td style="width:227px;">Carl Roth</td>
			<td style="width:259px;">HN68.3</td>
			<td>DOTA-conjugation</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:292px;">Ammonium acetate</td>
			<td style="width:227px;">Sigma-Aldrich</td>
			<td>32301</td>
			<td>DOTA-conjugation</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:292px;">PBS</td>
			<td style="width:227px;">University Tuebingen</td>
			<td style="width:259px;"></td>
			<td>DOTA-conjugation</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:292px;">Micro Bio-spin P-6 column</td>
			<td style="width:227px;">Bio-Rad Laboratories</td>
			<td style="width:259px;">7326221</td>
			<td>DOTA-conjugation</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:292px;">Sodium citrate</td>
			<td style="width:227px;">Sigma-Aldrich</td>
			<td style="width:259px;">71497</td>
			<td>DOTA-conjugation</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:292px;">Cyclone Plus PhosphorImager&#160;</td>
			<td style="width:227px;">Perkin-Elmer</td>
			<td>L2250116</td>
			<td>DOTA-conjugation</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:292px;">DMEM</td>
			<td style="width:227px;">Merck Millipore</td>
			<td>102568</td>
			<td>ingredient for T cell medium&#160;</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:292px;">FCS</td>
			<td style="width:227px;">Merck Millipore</td>
			<td style="width:259px;">S0115/1004B</td>
			<td>ingredient for T cell medium&#160;</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:292px;">Sodium pyruvate</td>
			<td style="width:227px;">Merck Millipore</td>
			<td style="width:259px;">L0473</td>
			<td>ingredient for T cell medium&#160;</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:292px;">MEM-amino acids</td>
			<td style="width:227px;">Merck Millipore</td>
			<td style="width:259px;">K0293</td>
			<td>ingredient for T cell medium&#160;</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:292px;">HEPES&#160;</td>
			<td style="width:227px;">Merck Millipore</td>
			<td style="width:259px;">L 1613</td>
			<td>ingredient for T cell medium&#160;</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:292px;">&#160;Penicillin/Streptomycin</td>
			<td style="width:227px;">Merck Millipore</td>
			<td style="width:259px;">A2212</td>
			<td>ingredient for T cell medium&#160;</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:292px;">0.05 mM 2-&#946;-mercaptoethanol</td>
			<td style="width:227px;">Sigma-Aldrich</td>
			<td style="width:259px;">M3148</td>
			<td>ingredient for T cell medium&#160;</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:292px;">DO11.10 mice</td>
			<td style="width:227px;"></td>
			<td>in-house breeding</td>
			<td>TH1 cell culture</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:292px;">DPBS</td>
			<td style="width:227px;">Gibco</td>
			<td style="width:259px;">14190144</td>
			<td>TH1 cell culture</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:292px;">Cell strainer 40 &#181;m&#160;</td>
			<td style="width:227px;">Corning</td>
			<td style="width:259px;">352340</td>
			<td>TH1 cell culture</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:292px;">ACK Lysing Buffer</td>
			<td style="width:227px;">Lonza</td>
			<td style="width:259px;">10-548E</td>
			<td>TH1 cell culture</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:292px;">CD4 MicroBeads, mouse</td>
			<td style="width:227px;">Miltenyi Biotech</td>
			<td style="width:259px;">130-097-145</td>
			<td>TH1 cell culture</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:292px;">QuadroMACS separator</td>
			<td style="width:227px;">Miltenyi Biotech</td>
			<td style="width:259px;">130-090-976</td>
			<td>TH1 cell culture</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:292px;">LS column</td>
			<td style="width:227px;">Miltenyi Biotech</td>
			<td style="width:259px;">130-042-401</td>
			<td>TH1 cell culture</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:292px;">anti-CD4 antibody (Gk1.5)</td>
			<td style="width:227px;"></td>
			<td>Isolated from hybridoma cell culture</td>
			<td>TH1 cell culture</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:292px;">anti-CD8 antibody (5367.2)</td>
			<td style="width:227px;"></td>
			<td>Isolated from hybridoma cell culture</td>
			<td>TH1 cell culture</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:292px;">Anti-rat antibody (MAR18.5)</td>
			<td style="width:227px;"></td>
			<td>Isolated from hybridoma cell culture</td>
			<td>TH1 cell culture</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:292px;">Rabbit complement MA</td>
			<td style="width:227px;">tebu-Bio</td>
			<td style="width:259px;">CL3221</td>
			<td>TH1 cell culture</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:292px;">Anti-IL-4 antibody (11B11)</td>
			<td style="width:227px;"></td>
			<td>Isolated from hybridoma cell culture</td>
			<td>TH1 cell culture</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:292px;">cOVA 323-339-peptide&#160;</td>
			<td style="width:227px;">EMC-micro-collections</td>
			<td>Custom order</td>
			<td>TH1 cell culture</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:292px;">CPG1668-oligonucleotides</td>
			<td style="width:227px;">Eurofins MWG Operon</td>
			<td>Custom order</td>
			<td>TH1 cell culture</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:292px;">IL-2</td>
			<td style="width:227px;">Novartis</td>
			<td>65483-116-07</td>
			<td>TH1 cell culture</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:292px;">96-well plates</td>
			<td style="width:227px;">Greiner&#160;</td>
			<td>655180</td>
			<td>TH1 cell culture</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:292px;">24-well plates</td>
			<td style="width:227px;">Greiner&#160;</td>
			<td>662160</td>
			<td>TH1 cell culture</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:292px;">cell culture flask</td>
			<td style="width:227px;">Greiner&#160;</td>
			<td>660175</td>
			<td>TH1 cell culture</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:292px;">48-well plates</td>
			<td style="width:227px;">Greiner&#160;</td>
			<td>677 180</td>
			<td>cell labeling</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:292px;">Gammacell 1000</td>
			<td style="width:227px;">Best Theratronics</td>
			<td>via inquiry&#160;</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Gulmay RT225</td>
			<td style="width:227px;">Gulmay</td>
			<td>via inquiry&#160;</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:292px;">Trypan blue</td>
			<td style="width:227px;">Merck Millipore</td>
			<td style="width:259px;">L6323</td>
			<td>in vitro evaluation</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:292px;">Mouse IFN-&#947; ELISA</td>
			<td style="width:227px;">BD Biosciences</td>
			<td style="width:259px;">558258</td>
			<td>in vitro evaluation</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:292px;">PE Annexin V Apoptosis Detection Kit&#160;</td>
			<td style="width:227px;">BD Biosciences</td>
			<td style="width:259px;">559763</td>
			<td>in vitro evaluation</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:292px;">Tube 5 ml</td>
			<td style="width:227px;">Sarstedt</td>
			<td style="width:259px;">55.476</td>
			<td>in vitro evaluation</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:292px;">Round-bottom tubes&#160;</td>
			<td style="width:227px;">BD Biosciences</td>
			<td style="width:259px;">352008</td>
			<td>in vitro evaluation</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:292px;">Wizard &#947;-counter</td>
			<td style="width:227px;">Perkin-Elmer</td>
			<td>2480-0010</td>
			<td>in vitro evaluation</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:292px;">ELISA Reader MultiscanEX</td>
			<td style="width:227px;">Thermo Fisher Scientific</td>
			<td>51118177</td>
			<td>in vitro evaluation</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:292px;">Microscope</td>
			<td style="width:227px;">Leica</td>
			<td>via inquiry&#160;</td>
			<td>in vitro evaluation</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:292px;">BD LSRII&#160;</td>
			<td style="width:227px;">BD Biosciences</td>
			<td>via inquiry&#160;</td>
			<td>in vitro evaluation</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:292px;">BALB/c mice</td>
			<td style="width:227px;">Charles River</td>
			<td style="width:259px;">028</td>
			<td>in vivo cell trafficking</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:292px;">Aluminum gel</td>
			<td style="width:227px;">Serva Electrophoresis</td>
			<td style="width:259px;">12261.01</td>
			<td>in vivo cell trafficking</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:292px;">Xylazine</td>
			<td style="width:227px;">Bayer HealthCare</td>
			<td>Ordered via University hospital</td>
			<td>in vivo cell trafficking</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:292px;">Ketamine</td>
			<td style="width:227px;">Ratiopharm</td>
			<td>Ordered via University hospital</td>
			<td>in vivo cell trafficking</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:292px;">Isoflurane</td>
			<td style="width:227px;">CP-Pharma</td>
			<td>Ordered via University hospital</td>
			<td>in vivo cell trafficking</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:292px;">30G needle</td>
			<td style="width:227px;">BD Biosciences</td>
			<td>304000</td>
			<td>in vivo cell trafficking</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:292px;">Syringe</td>
			<td style="width:227px;">BD Biosciences</td>
			<td>11612491</td>
			<td>in vivo cell trafficking</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:292px;">Capillaries 10 &#181;l</td>
			<td style="width:227px;">VWR</td>
			<td>612-2439</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:292px;">Inveon PET scanner</td>
			<td style="width:227px;">Siemens Healthineers</td>
			<td style="width:259px;">no longer available</td>
			<td>in vivo cell trafficking, alternative companies: Bruker, Mediso&#160;</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:292px;">Inveon SPECT/CT scanner</td>
			<td style="width:227px;">Siemens Healthineers</td>
			<td style="width:259px;">no longer available</td>
			<td>in vivo cell trafficking</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:292px;">Inveon Research Workplace</td>
			<td style="width:227px;">Siemens Healthineers</td>
			<td style="width:259px;"></td>
			<td>image analysis, alternative software: Pmod</td>
		</tr>
	</tbody>
</table>

murine lymphocyte labeling by 64cu-antibody receptor targeting for in vivo cell trafficking by pet/ct

Bu protokol, 64Cu üretimini ve kemirgen lenfosit hücre kültürü ve hücre hedefleme 64Cu-reseptörün ardından bir monoklonal antikor (mAb) şelatlayıcı çekimi / Radyo-etiketlenmesi göstermektedir. PET / CT ile bir hava yolu gecikmiş tip aşın duyarlılık reaksiyonunda (DTHR) bir hayvan modelinde radyo-etiket, ve in vivo hücre takibi yayılmayan açıklanmaktadır in vitro değerlendirilmesi. Ayrıntılı olarak, şelatlayıcı 1,4,7,10-tetraazasiklododekan-1,4,7,10-tetraasetik asit (DOTA) bir mAb konjugasyon gösterilmiştir. Radyoaktif 64 Cu, DOTA-konjuge mAb&#39;nin radyoaktif üretimini sonra tarif edilmektedir. Daha sonra, tavuk ovalbumin (cova) e özgü CD4 + interferon genişlemesi (IFN) -γ-üreten T yardımcı hücreleri (cova-TH1) ve cova-TH1 hücrelerinin daha sonra, radyo-etiketleme olarak gösterilmiştir. Çeşitli in vitro teknikler ef değerlendirmek için sunulmuşturtripan mavi dışlama hücre canlılığının belirlenmesi, akış sitometrisi için Annexin V ile apoptosis için boyama ve IFN-γ enzime bağlı immünosorban tahliliyle özelliklerin değerlendirilmesi gibi hücreler üzerinde 64Cu-radyo sınıflandırılmasıyla tama yaptığı, (ELISA) . Bundan başka, hücre içine radyoaktivite alımının ve markalama kararlılığının belirlenmesi detaylı olarak tarif edilmektedir. Bu protokol daha nedenle, BALB / c farelerinde cova kaynaklı akut solunum yolu DHTR indüksiyonu dahil, bir hava yolu DTHR için bir hayvan modeli olarak, hücre izleme çalışmaları yapmak ve açıklamaktadır. Son olarak, görüntü elde etme, yeniden yapılanma, ve analizi dahil sağlam PET / BT iş akışı sunulmuştur. Daha sonra alıcı içselleştirme 64 Cu-antikor reseptörü hedefleyen bir yaklaşım, hücre etiketlenmesi için ortak PET izleyiciler kıyasla yüksek spesifite ve stabilite, düşük hücre toksisitesi ve düşük akış oranlarını sağlar, örneğin, 64Cu-pyruvaldehyde bis (N-4-methylthiosemicarbazone) (64Cu-PTSM). Son olarak, yaklaşım, 48 saat için optimum sinyal-zemin oranı PET / CT ile in vivo hücre takibi yayılmayan sağlar. Bu deneysel yaklaşım içselleştirilir membrana bağlı reseptör ile hayvan modelleri ve hücre tipleri farklı transfer edilebilir.

Şekil 1, 64, Cu-DOTA-KJ1-26-mAb ve in vitro ve bu protokol kaplı vivo çalışmalarda deney tasarımı ile cova-TH1 hücrelerinin etiketlenmesi özetlemektedir. <img alt="Şekil 1" src="/files/ftp_upload/55270/55270fig1.jpg" /> Şekil 1: 64 Cu-DOTA-KJ1-26 mAb Etiketleme Proses ve Deney Tasarımı. 64Cu-DOTA-KJ1-26-mAb ile radyoaktif eti...

Watch this Scientific Journal Video about Murine Lymphocyte Labeling by 64Cu-Antibody Receptor Targeting for In Vivo  Cell Trafficking by PET/CT at JoVE.com

Murine Lymphocyte Labeling by 64Cu-Antibody Receptor Targeting for In Vivo  Cell Trafficking by PET/CT

Bir transgenik fare, T hücresi reseptörüne bağlanan bir 64Cu-modifiye edilmiş monoklonal antikorun hazırlanmasını takiben, T hücreleri, canlılık, fonksiyonellik, etiketleme stabilitesi ve apoptoz için analiz, in vivo olarak radyoaktif işaretli olan ve adoptif bir nefes yolu gecikmeli tipte aşırı duyarlılık ile farelere aktarıldı pozitron emisyon tomografisi / bilgisayarlı tomografi (PET / BT) ile invazif olmayan görüntüleme için reaksiyon.

tarafından Kemirgen Lenfosit Etiketleme<sup&gt; 64</sup&gt; Cu-Antikor Reseptör için hedefleme<em&gt; In Vivo</em&gt; PET / CT ile hücre Trafik

This protocol illustrates the production of 64Cu and the chelator conjugation/radiolabeling of a monoclonal antibody (mAb) followed by murine lymphocyte ...

Research

JoVE Journal

Immunology and Infection

Murine Lymphocyte Labeling by 64Cu-Antibody Receptor Targeting for In Vivo Cell Trafficking by PET/CT

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

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

Hücre-kaderi ardından<em&gt; E. coli</em&gt; Faj Lambda tarafından Enfeksiyon Sonrası

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

Induction of Murine Intestinal Inflammation by Adoptive Transfer of Effector CD4+CD45RBhigh T Cells into Immunodeficient Mice

Induction of Murine Intestinal Inflammation by Adoptive Transfer of Effector CD4+CD45RBhigh T Cells into Immunodeficient Mice

Efektör CD4 Evlatlık transferi ile Kemirgen Bağırsak Enflamasyon indüksiyonu<sup&gt; +</sup&gt; CD45RB<sup&gt; Yüksek</sup&gt; Bağışık yetmezliği olan farelere içine T Hücreleri

There are many different animal models available for studying the pathogenesis of human inflammatory bowel diseases (IBD), each with its own advantages ...

Lymphocyte Isolation from Human Skin for Phenotypic Analysis and Ex Vivo Cell Culture

Lymphocyte Isolation from Human Skin for Phenotypic Analysis and Ex Vivo Cell Culture

Fenotipik Analizi ve İnsan Cilt lenfosit izolasyonu<em&gt; Ex Vivo</em&gt; Hücre Kültürü

Human skin has an important barrier function and contains various immune cells that contribute to tissue homeostasis and protection from pathogens. As the ...

Femur Window Chamber Model for In Vivo Cell Tracking in the Murine Bone Marrow

Femur Window Chamber Model for In Vivo Cell Tracking in the Murine Bone Marrow

için Femur Pencere Odası Modeli<em&gt; İn Vivo</emFare Kemik İliği içinde&gt; Hücre Takibi

Bone marrow is a complex organ that contains various hematopoietic and non-hematopoietic cells. These cells are involved in many biological processes, ...

Detection of Trypanosoma brucei Variant Surface Glycoprotein Switching by Magnetic Activated Cell Sorting and Flow Cytometry

Detection of Trypanosoma brucei Variant Surface Glycoprotein Switching by Magnetic Activated Cell Sorting and Flow Cytometry

Algılama<em&gt; Trypanosoma brucei</emManyetik Aktif Hücre Sorting ve Akış Sitometrisi ile&gt; Varyant Yüzey Glikoprotein Değiştirme

Trypanosoma brucei, a protozoan parasite that causes both Human and Animal African Trypanosomiasis (known as sleeping sickness and nagana, respectively) ...

Detection of Enterohemorrhagic Escherichia Coli Colonization in Murine Host by Non-invasive In Vivo Bioluminescence System

Detection of Enterohemorrhagic Escherichia Coli Colonization in Murine Host by Non-invasive In Vivo Bioluminescence System

Non-invaziv Vivo içinde Bioluminescence sistemi tarafından fare konak Enterohemorrhagic Escherichia Coli kolonizasyon algılama

Enterohemorrhagic E. coli (EHEC) O157:H7, which is a foodborne pathogen that causesdiarrhea, hemorrhagic colitis (HS), and hemolytic uremic syndrome ...

Quantifying Human Monocyte Chemotaxis In Vitro and Murine Lymphocyte Trafficking In Vivo

Quantifying Human Monocyte Chemotaxis In Vitro and Murine Lymphocyte Trafficking In Vivo

İnsan monosit kemotaksisi Vitro ve In Vivo kaçakçılığı fare lenfosit miktarının

Chemotaxis is migration along a specific chemical gradient1. Chemokines are chemotactic cytokines that promote cellular trafficking with anatomic and ...

Analysis of 18FDG PET/CT Imaging as a Tool for Studying Mycobacterium tuberculosis Infection and Treatment in Non-human Primates

Analysis of 18FDG PET/CT Imaging as a Tool for Studying Mycobacterium tuberculosis Infection and Treatment in Non-human Primates

Analiz 18FDG PET/BT uzmanı Mycobacterium tüberküloz enfeksiyon ve insan olmayan primatlarda tedavisi için bir araç olarak görüntüleme

Mycobacterium tuberculosis remains the number one infectious agent in the world today. With the emergence of antibiotic resistant strains, new clinically ...

Visualizing Intracellular SNARE Trafficking by Fluorescence Lifetime Imaging Microscopy

Hücre içi SNARE mikroskobu Imaging Floresans ömür tarafından kaçakçılığı görüntülenmesi

Soluble N-ethylmaleimide sensitive fusion protein (NSF) attachment protein receptor (SNARE) proteins are key for membrane trafficking, as they catalyze ...

Assessment of Lymphocyte Migration in an Ex Vivo Transmigration System

Ex Vivo Transmigration Sisteminde Lenfosit Göçünün Değerlendirilmesi

Herein, we present an efficient method that can be executed with basic laboratory skills and materials to assess lymphocyte chemokinetic movement in an ex ...