저자는 실험 및 데이터 분석시 지원을 위해 박사 줄리아 만하임, 월터 에를리치만, 라모나 스텀, Funda 케이, 다니엘 부칼라, 마렌 하란트뿐만 아니라 나탈리 Altmeyer 감사합니다. 이 작품은 베르너 지멘스-재단, SFB685 통해 DFG (프로젝트 B6)과 재산 (2309-0-0)에 의해 지원되었다.

안전주의 사항 : 방사능을 처리 할 때, 2 인치 두께의 납 벽돌 뒤에 (64) 구리를 저장하고 활동을 운반하는 모든 선박에 대한 각각의 차폐를 사용합니다. 간접적으로 직접 손 접촉을 방지하고 방사성 물질에 대한 노출을 최소화하기 위해 차폐 소스를 처리하기 위해 적절한 도구를 사용하십시오. 항상 방사선 선량 모니터링 배지 및 개인 보호 장비를 착용하고 자신을 확인하고 오염의 작업 ?...

<ol>
	<li>Cerri, 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=Intracarotid+Infusion+of+Mesenchymal+Stem+Cells+in+an+Animal+Model+of+Parkinson's+Disease,+Focusing+on+Cell+Distribution+and+Neuroprotective+and+Behavioral+Effects.">Intracarotid Infusion of Mesenchymal Stem Cells in an Animal Model of Parkinson's Disease, Focusing on Cell Distribution and Neuroprotective and Behavioral Effects.</a> Stem Cells Trans Med. 4 (9), 1073-1085 (2015).</li><li>Hasenbach, K., 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=Monitoring+the+glioma+tropism+of+bone+marrow-derived+progenitor+cells+by+2-photon+laser+scanning+microscopy+and+positron+emission+tomography.">Monitoring the glioma tropism of bone marrow-derived progenitor cells by 2-photon laser scanning microscopy and positron emission tomography.</a> Neuro Oncol. 14 (4), 471-481 (2012).</li><li>Sood, V., 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=Biodistribution+of+18F-FDG-Labeled+Autologous+Bone+Marrow+&#8211;+Derived+Stem+Cells+in+Patients+With+Type+2+Diabetes+Mellitus.">Biodistribution of 18F-FDG-Labeled Autologous Bone Marrow &#8211; Derived Stem Cells in Patients With Type 2 Diabetes Mellitus.</a> Clin Nucl Med. 40 (9), 697-700 (2015).</li><li>Perez-Diez, A., 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=CD4+cells+can+be+more+efficient+at+tumor+rejection+than+CD8+cells.">CD4 cells can be more efficient at tumor rejection than CD8 cells.</a> Blood. 109 (12), 5346-5354 (2007).</li><li>Muranski, P., Restifo, N. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Adoptive+immunotherapy+of+cancer+using+CD4++T+cells.">Adoptive immunotherapy of cancer using CD4+ T cells.</a> Curr. Opin. Immunol. 21 (2), 200-208 (2009).</li><li>Braumuller, 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=T-helper-1-cell+cytokines+drive+cancer+into+senescence.">T-helper-1-cell cytokines drive cancer into senescence.</a> Nature. 494 (7437), 361-365 (2013).</li><li>Kochenderfer, J. 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=Eradication+of+B-lineage+cells+and+regression+of+lymphoma+in+a+patient+treated+with+autologous+T+cells+genetically+engineered+to+recognize+CD19.">Eradication of B-lineage cells and regression of lymphoma in a patient treated with autologous T cells genetically engineered to recognize CD19.</a> Blood. 116 (20), 4099-4102 (2010).</li><li>Cherry, S. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fundamentals+of+Positron+Emission+Tomography+and+Applications+in+Preclinical+Drug+Development.">Fundamentals of Positron Emission Tomography and Applications in Preclinical Drug Development.</a> J. Clin. Pharmacol. 41 (5), 482-491 (2001).</li><li>Tavar&#233;, 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=An+Effective+Immuno-PET+Imaging+Method+to+Monitor+CD8-Dependent+Responses+to+Immunotherapy.">An Effective Immuno-PET Imaging Method to Monitor CD8-Dependent Responses to Immunotherapy.</a> Cancer Res. 76 (1), 73-82 (2016).</li><li>Tavar&#233;, 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=Engineered+antibody+fragments+for+immuno-PET+imaging+of+endogenous+CD8++T+cells+in+vivo.">Engineered antibody fragments for immuno-PET imaging of endogenous CD8+ T cells in vivo.</a> Proc. Natl. Acad. Sci. U.S.A. 111 (3), 1108-1113 (2014).</li><li>Dobrenkov, K., 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=Monitoring+the+Efficacy+of+Adoptively+Transferred+Prostate+Cancer-Targeted+Human+T+Lymphocytes+with+PET+and+Bioluminescence+Imaging.">Monitoring the Efficacy of Adoptively Transferred Prostate Cancer-Targeted Human T Lymphocytes with PET and Bioluminescence Imaging.</a> J Nucl Med. 49 (7), 1162-1170 (2008).</li><li>Rini, J. 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=PET+with+FDG-labeled+Leukocytes+versus+Scintigraphy+with+111In-Oxine-labeled+Leukocytes+for+Detection+of+Infection.">PET with FDG-labeled Leukocytes versus Scintigraphy with 111In-Oxine-labeled Leukocytes for Detection of Infection.</a> Radiology. 238 (3), 978-987 (2006).</li><li>Ritchie, D., 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=In+vivo+tracking+of+macrophage+activated+killer+cells+to+sites+of+metastatic+ovarian+carcinoma.">In vivo tracking of macrophage activated killer cells to sites of metastatic ovarian carcinoma.</a> Cancer Immunol. Immunother. 56 (2), 155-163 (2006).</li><li>Adonai, 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=Ex+vivo+cell+labeling+with+64Cu-pyruvaldehyde-bis(N4-methylthiosemicarbazone)+for+imaging+cell+trafficking+in+mice+with+positron-emission+tomography.">Ex vivo cell labeling with 64Cu-pyruvaldehyde-bis(N4-methylthiosemicarbazone) for imaging cell trafficking in mice with positron-emission tomography.</a> Proc. Natl. Acad. Sci. U.S.A. 99 (5), 3030-3035 (2002).</li><li>Huang, J., Lee, C. C. I., Sutcliffe, J. L., Cherry, S. R., Tarantal, A. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Radiolabeling+Rhesus+Monkey+CD34++Hematopoietic+and+Mesenchymal+Stem+Cells+with+64Cu-Pyruvaldehyde-Bis(N4-Methylthiosemicarbazone)+for+MicroPET+Imaging.">Radiolabeling Rhesus Monkey CD34+ Hematopoietic and Mesenchymal Stem Cells with 64Cu-Pyruvaldehyde-Bis(N4-Methylthiosemicarbazone) for MicroPET Imaging.</a> Mol. Imaging. 7 (1), (2008).</li><li>Griessinger, C. 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=In+Vivo+Tracking+of+Th1+Cells+by+PET+Reveals+Quantitative+and+Temporal+Distribution+and+Specific+Homing+in+Lymphatic+Tissue.">In Vivo Tracking of Th1 Cells by PET Reveals Quantitative and Temporal Distribution and Specific Homing in Lymphatic Tissue.</a> J Nucl Med. 55 (2), 301-307 (2014).</li><li>Griessinger, C. 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=64Cu+antibody-targeting+of+the+T-cell+receptor+and+subsequent+internalization+enables+in+vivo+tracking+of+lymphocytes+by+PET.">64Cu antibody-targeting of the T-cell receptor and subsequent internalization enables in vivo tracking of lymphocytes by PET.</a> Proc. Natl. Acad. Sci. U.S.A. 112 (4), 1161-1166 (2015).</li><li>McCarthy, D. 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=Efficient+production+of+high+specific+activity+64Cu+using+a+biomedical+cyclotron.">Efficient production of high specific activity 64Cu using a biomedical cyclotron.</a> Nucl. Med. Biol. 24 (1), 35-43 (1997).</li><li>Kalkhof, S., Sinz, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chances+and+pitfalls+of+chemical+cross-linking+with+amine-reactive+N-hydroxysuccinimide+esters.">Chances and pitfalls of chemical cross-linking with amine-reactive N-hydroxysuccinimide esters.</a> Anal. Bioanal. Chem. 392 (1), 305-312 (2008).</li><li>Bedoya, S. K., Wilson, T. D., Collins, E. L., Lau, K., Larkin Iii, ., J, <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Isolation+and+Th17+Differentiation+of+Naive+CD4+T+Lymphocytes.">Isolation and Th17 Differentiation of Naive CD4 T Lymphocytes.</a> J Vis Exp. (79), e50765 (2013).</li><li>Flaherty, S., Reynolds, J. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mouse+Naive+CD4++T+Cell+Isolation+and+In+vitro+Differentiation+into+T+Cell+Subsets.">Mouse Naive CD4+ T Cell Isolation and In vitro Differentiation into T Cell Subsets.</a> J Vis Exp. (98), e52739 (2015).</li><li>Judenhofer, M., Wiehr, S., Kukuk, D., Fischer, K., Pichler, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chapter+363.">Chapter 363.</a> Small Animal Imaging. Basics and Practical Guide. , 363-370 (2011).</li><li>Phelps, M. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> PET. Molecular Imaging and Its Biological Applications. , 93-101 (2004).</li><li>Wu, A. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Antibodies+and+Antimatter:+The+Resurgence+of+Immuno-PET.">Antibodies and Antimatter: The Resurgence of Immuno-PET.</a> JNM. 50 (1), 2-5 (2009).</li><li>Lewis, M. 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=In+vivo+evaluation+of+pretargeted+64Cu+for+tumor+imaging+and+therapy.">In vivo evaluation of pretargeted 64Cu for tumor imaging and therapy.</a> J Nucl Med. 44 (8), 1284-1292 (2003).</li><li>Boswell, C. A., 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+in+vivo+stability+of+copper-64-labeled+cross-bridged+and+conventional+tetraazamacrocyclic+complexes.">Comparative in vivo stability of copper-64-labeled cross-bridged and conventional tetraazamacrocyclic complexes.</a> J Med Chem. 47 (6), 1465-1474 (2004).</li><li>Ghosh, S. 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=Comparison+of+DOTA+and+NODAGA+as+chelators+for+(64)Cu-labeled+immunoconjugates.">Comparison of DOTA and NODAGA as chelators for (64)Cu-labeled immunoconjugates.</a> Nucl Med Biol. 42 (2), 177-183 (2015).</li><li>Johnson, T. E., Birky, B. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Health Physics and Radiological Health. , (2011).</li><li>Helmchen, F., Denk, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Deep+tissue+two-photon+microscopy.">Deep tissue two-photon microscopy.</a> Nat Meth. 2 (12), 932-940 (2005).</li></ol>

이 프로토콜은 안정적으로 PET에 의해 생체 내 추적 세포를 방사성 표지하는 신뢰할 수있는 쉬운 방법을 제시한다. 64 CU-DOTA-KJ1-26 - 단클론 항체 및 그 유도와 방사성 표지 수, 코바-TH1 세포, 절연 및 기증자 마우스의 체외에서 확장이 방법을 수 활용하는 A의 코바 프레젠테이션의 사이트로 폐와 perithymic 림프절을 추적했다 급성기도 DTHR을 COVA 유도. 킬레이?...

비 침습적 셀 추적은 생체 내에서 세포 기능, 이동과 원점을 모니터링하는 다용도 공구이다. 최근 세포 추적 연구는 재생 의학, 암 3, 4에 대한 입양 세포 치료에 염증이나 T 림프구에서자가 말초 백혈구의 맥락에서 중간 엽 1, 2 또는 골수 유래 줄기 세포 3에 초점을 맞추고있다. 행동의 사이트 및 세포 기반 치료의 기본 생물학적 원리의 해명은 엄청난 중요하다. CD8 + 세포 독성 T 림프구는 유전자 널리 금 표준으로 간주 하였다 키메라 항원 수용체 (CAR) T 세포 또는 종양 침윤 림프구 (TILS)를 설계. 그러나, 종양 관련 항원 특이 TH1 세포는 &lt;효과적인 대안 치료 옵션 4, 입증했다/ SUP&gt; 5, 6, 7. 키 염증 플레이어, 기관 고유의자가 면역 질환 (예를 들어, 류마티스 관절염 또는 기관지 천식), 암 면역 요법에 높은 관심의 세포, TH1 세포의 시간적 분포와 유도 패턴을 특성화하는 것이 중요하다. PET에 의한 비 침습적 생체 이미징 생체 원위치에서 세포의 이동 패턴을 조사하는 계량 고도로 민감한 방법 (8)을 제공하고, 염증, 알레르기, 감염 또는 종양 거부 9, 10, 11 중 T 세포의 작용과 반응 사이트. 임상 적으로, 인 옥신 (111)는 2- 데 옥시 -2- (18 동안, 염증 및 감염 (12)의 판정에 대한 백혈구 신티에 사용F) 플루오로 -D- 글루코스 (FDG-18 F)는 일반적으로 PET (3), 셀 (13)에 의해 추적 연구에 사용된다. 이 PET 추적자 하나의 큰 단점은, 그러나, 109.7 분의 핵종 18 F 이후 시점에서 촬상이 대체 세포 이동을 저해 게시 낮은 세포 내 안정성의 짧은 반감기이다. 세포 불안정하지만, 64의 Cu-PTSM 빈번하게하는 데 사용되는 PET 생체 내 세포 추적 연구에서 장기에 대해 비특이적 전지 (14), T 세포 생존에 최소화 해로운 효과 (15)에 라벨 (16) 기능을한다. 이 프로토콜은 또한 T 세포 수용체 (TCR) 특이 방사성 표지 된 항체를 이용하여 세포 생존 및 기능에 대한 불리한 영향을 줄이는 방법을 설명한다. 우선, 방사성 동위 원소 64의 Cu, 제 KJ1-26와 단클론 항체의 결합의 제조E DOTA 킬레이트 및 후속 CU-64 방사성 표지를 나타낸다. 제 2 단계에서, 상기 절연 및 DO11.10 공여 마우스의 코바-TH1 세포의 확장 및 64의 Cu-DOTA로드 공역 단클론 KJ1-26 (64 CU-DOTA-KJ1-26)와 방사성 표지를 상세히 설명한다. 흡수 값 및 도즈 교정과 방사능 유출과 각각 감마 계수에 의해 평가뿐만 아니라, IFN-γ ELISA가 제시와 트립 판 블루 배제 및 기능에 의한 세포 생존율의 64 CU-방사성 표지의 효과의 평가 . 생체 세포의 추적 비 침습적 들어, 대체 세포 전사 후에 PET / CT 의해 COVA 유발 급성기도 DTHR 및 이미지 획득의 마우스 모델의 도출을 설명한다. 또한,이 라벨 접근 방식은 다른 질병 모델, 다른 TCR도 또는 막 결합 수용체 또는 식 마르와 관심의 일반 세포와 쥐의 T 세포로 전송할 수 있습니다KERS 연속 막 (17)을 왕복 밑에.

<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

이 프로토콜은 구리 (64)의 제조 및 뮤린 림프구 세포 배양 및 세포 타겟팅 CU-64 수용체 항체에 따르는 단일 클론 항체 (항체)의 킬레이트 결합 / 방사성 표지를 나타낸다. PET / CT에 의한기도 지연 형 과민 반응 (DTHR)의 동물 모델에서 생체 세포와 방사성 추적 비 침습적 설명되는 시험 관내 평가. 구체적으로는, 킬레이트 1,4,7,10-1,4,7,10 테트라 아자 - 테트라 아세트산 (DOTA)와 항체의 결합을 나타낸다. 방사선 (64)의 Cu의 DOTA 공역의 방사성 표지 된 단클론 항체의 생산을하는 것은 다음을 설명한다. 다음에, 닭 오브 알부민 (코바) 특이 적 CD4 + 인터페론의 확장 (IFN)의 생산 -γ-T 헬퍼 세포 (COVA-TH1)과 코바-TH1 세포의 후속의 방사선은 도시된다. 다양한 체외 기술은 EF을 평가하기 위해 제시이러한 트리 판 블루 배제에 의한 세포 생존의 판정, 유동 세포 계측법 넥신 V와 아폽토시스에 대한 염색 및 IFN-γ 효소 면역 분석법에 의해 기능의 평가와 셀 64의 Cu-방사성 표지의 fects (ELISA) . 또한, 세포 내로 흡수하고 방사성 표지 안정성의 판정을 상세히 설명한다. 이 프로토콜은 상기 따라서, BALB / c 마우스에서 코바 유발 급성기도 DHTR의 유도가 포함되어기도 DTHR에 대한 동물 모델에서 세포 추적 연구를 수행하는 방법을 설명한다. 마지막으로, 이미지 획득, 재구성, 및 분석을 포함하는 강력 PET / CT 흐름이 제공된다. 후속 수용체 내재화와 CU-64 수용체 항체 타겟팅 방법은 셀 공통 표지 용 PET-트레이서에 비해 높은 안정성 및 특이성 감소 세포 독성, 낮은 유출 속도를 제공 예 64의 Cu-pyruvaldehyde 비스 (N4-methylthiosemicarbazone) (64 CU-PTSM). 마지막으로, 우리의 방법은 48 시간을위한 최적의 신호 대 백그라운드 비율 PET / CT 생체 내 세포 추적 비 침습적 수있다. 이러한 실험 방법은 내재화되는 막 결합 수용체와 동물 모델과 다른 종류의 세포에 전달 될 수있다.

도 1은 64의 Cu-DOTA-KJ1-26 단클론 및 시험 관내 및이 프로토콜에 덮여 생체 내 연구에서의 실험 디자인 COVA-TH1 세포의 표지를 요약 한 것이다. <img alt="그림 1" src="/files/ftp_upload/55270/55270fig1.jpg" /> 그림 1 : 64 CU-DOTA-KJ1-26 - 단클론 항체 라벨링 공정 및 실험 설계. 64 CU-...

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

형질 전환 쥐 T 세포 수용체에 결합하는 64의 Cu 변성 모노클로 날 항체의 제조에 이어 T 세포 생존, 기능, 표시 안정성과 세포 사멸 분석, 생체 내에서 방사성 표지하고, 적응 적기도 지연 형 과민 쥐에 옮기고 양전자 방출 단층 촬영 / 컴퓨터 단층 촬영 (PET / CT)에 의해 비 침습성 영상화 대한 반응.

에 의해 쥐 림프구 라벨링<sup&gt; (64)</sup&gt; CU-항체 수용체는 대한 타겟팅<em&gt; 생체</em&gt; PET / CT에 의한 세포 트래 피킹

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

에 셀 운명을 따라<em&gt; E. 대장균</em&gt; 파지 람다에 의해 감염 후

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

연구

면역학 및 감염병학

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

이펙터 CD4의 입양 전송하여 쥐과 장내 염증의 유도<sup&gt; +</sup&gt; CD45RB<sup&gt; 높은</sup&gt; 면역 결핍 마우스에게 T 세포

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

표현형 분석 및 대한 인간의 피부에서 림프구 분리<em&gt; 전의 VIVO</em&gt; 세포 배양

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

대한 대퇴골 창 회의소 모델<em&gt; 생체</em쥐의 골수에서&gt; 셀 추적

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

검출<em&gt; 트리파노소마 (Trypanosoma) brucei</em자기 활성 셀 정렬 및 유동 세포 계측법에 의해&gt; 변형 표면 당 단백질 전환

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

생물 발광 시스템 비-침략 적 비보에 의해 Murine 호스트에서 Enterohemorrhagic 대장균 식민지의 검출

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

인간 Monocyte Chemotaxis에서 생체 외에서 그리고 Vivo에서 인신 매매 Murine 림프 구 측정

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

18FDG PET/CT 영상 공부 결핵균 감염 및 비 인간 영장류에서 치료 도구로 서의 분석

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

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 트랜스이민 시스템에서 림프구 마이그레이션 평가

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