Name: intracellular staining and flow cytometry to identify lymphocyte subsets within murine aorta, kidney and lymph nodes in a model of hypertension
Uploaded: 2017-01-28T13:00:00
Description: Watch this Scientific Journal Video about Intracellular Staining and Flow Cytometry to Identify Lymphocyte Subsets within Murine Aorta, Kidney and Lymph Nodes in a Model of Hypertension at JoVE.com

Este trabalho foi apoiado por uma concessão do American Heart Association Fellowship (16POST29950007) para FL, uma bolsa de formação dos Institutos Nacionais de Saúde (NIH T32 HL069765) para BLD, um Prêmio Associação Fellowship American Heart (14POST20420025) para MA Saleh, e um NIH prêmio K08 (HL121671) para MSM. MSM também é apoiado por uma bolsa de investigação Gilead Sciences, Inc.

Comitê de Cuidado e Uso Institucional Animal da Universidade de Vanderbilt, aprovou os procedimentos aqui descritos. Os ratos são alojados e tratados de acordo com o Guia para o Cuidado e Uso de Animais de Laboratório (National Academies Press. Revised 2010). 1. Isolamento da aorta Drenagem Linfática Nodes, rim e aorta de ratos <ol><li> Eutanásia dos ratinhos por inalação de CO2. Pulveriza-se a caixa com 70% de etanol e abrir cuidadosamente a pele e a parede do tórax com uma tesoura para exp...

<ol>
	<li>Saleh, M. A., McMaster, W. G., Wu, 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=Lymphocyte+adaptor+protein+LNK+deficiency+exacerbates+hypertension+and+end-organ+inflammation.">Lymphocyte adaptor protein LNK deficiency exacerbates hypertension and end-organ inflammation.</a> J Clin Invest. 125 (3), 1189-1202 (2015).</li><li>Madhur, M. S., Lob, H. E., McCann, L. 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=Interleukin+17+promotes+angiotensin+II-induced+hypertension+and+vascular+dysfunction.">Interleukin 17 promotes angiotensin II-induced hypertension and vascular dysfunction.</a> Hypertension. 55 (2), 500-507 (2010).</li><li>Laroumanie, F., Douin-Echinard, V., Pozzo, 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=CD4++T+Cells+Promote+the+Transition+From+Hypertrophy+to+Heart+Failure+During+Chronic+Pressure+Overload.">CD4+ T Cells Promote the Transition From Hypertrophy to Heart Failure During Chronic Pressure Overload.</a> Circulation. 129 (21), 2111-2124 (2014).</li><li>Guzik, T. J., Hoch, N. E., Brown, K. 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=Role+of+the+T+cell+in+the+genesis+of+angiotensin+II+induced+hypertension+and+vascular+dysfunction.">Role of the T cell in the genesis of angiotensin II induced hypertension and vascular dysfunction.</a> J Exp Med. 204 (10), 2449-2460 (2007).</li><li>Ketelhuth, D. F. J., Hansson, G. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Adaptive+Response+of+T+and+B+Cells+in+Atherosclerosis.">Adaptive Response of T and B Cells in Atherosclerosis.</a> Circ Res. 118 (4), 668-678 (2016).</li><li>McMaster, W. G., Kirabo, A., Madhur, M. S., Harrison, D. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Inflammation,+Immunity,+and+Hypertensive+End-Organ+Damage.">Inflammation, Immunity, and Hypertensive End-Organ Damage.</a> Circ Res. (6), 1022-1033 (2015).</li><li>Luckheeram, R. V., Zhou, R., Verma, A. 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=CD4+T+Cells:+Differentiation+and+Functions.">CD4+T Cells: Differentiation and Functions.</a> Clin Dev Immunol. 2012, 1-12 (2012).</li><li>Mittr&#252;cker, H. -. W., Visekruna, A., Huber, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Heterogeneity+in+the+differentiation+and+function+of+CD8++T+cells.">Heterogeneity in the differentiation and function of CD8+ T cells.</a> Arch Immunol Ther Exp (Warsz). 62 (6), 449-458 (2014).</li><li>Autengruber, A., Gereke, M., Hansen, G., Hennig, C., Bruder, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Impact+of+enzymatic+tissue+disintegration+on+the+level+of+surface+molecule+expression+and+immune+cell+function.">Impact of enzymatic tissue disintegration on the level of surface molecule expression and immune cell function.</a> Eur J Microbiol Immunol (Bp). 2 (2), 112-120 (2012).</li><li>Finak, G., Langweiler, M., Jaimes, 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=Standardizing+Flow+Cytometry+Immunophenotyping+Analysis+from+the+Human+ImmunoPhenotyping+Consortium.">Standardizing Flow Cytometry Immunophenotyping Analysis from the Human ImmunoPhenotyping Consortium.</a> Sci Rep. 6, 20686 (2016).</li><li>Herzenberg, L. A., Parks, D., Sahaf, B., Perez, O., Roederer, M., Herzenberg, L. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+history+and+future+of+the+fluorescence+activated+cell+sorter+and+flow+cytometry:+a+view+from+Stanford.">The history and future of the fluorescence activated cell sorter and flow cytometry: a view from Stanford.</a> Clin Chem. 48 (10), 1819-1827 (2002).</li><li>Hrvatin, S., Deng, F., O'Donnell, C. W., Gifford, D. K., Melton, D. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=MARIS:+method+for+analyzing+RNA+following+intracellular+sorting.">MARIS: method for analyzing RNA following intracellular sorting.</a> PLoS One. 9 (3), 89459 (2014).</li><li>Kalisky, T., Quake, 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=Single-cell+genomics.">Single-cell genomics.</a> Nat Methods. 8 (4), 311-314 (2011).</li><li>Chattopadhyay, P. K., Roederer, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cytometry:+Today's+technology+and+tomorrow's+horizons.">Cytometry: Today's technology and tomorrow's horizons.</a> Methods. 57 (3), 251-258 (2012).</li><li>Jiang, L., Tixeira, R., Caruso, 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=Monitoring+the+progression+of+cell+death+and+the+disassembly+of+dying+cells+by+flow+cytometry.">Monitoring the progression of cell death and the disassembly of dying cells by flow cytometry.</a> Nat Protoc. 11 (4), 655-663 (2016).</li><li>Lemoine, S., Jaron, B., Tabka, 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=Dectin-1+activation+unlocks+IL12A+expression+and+reveals+the+TH1+potency+of+neonatal+dendritic+cells.">Dectin-1 activation unlocks IL12A expression and reveals the TH1 potency of neonatal dendritic cells.</a> J Allergy Clin Immunol. 136 (5), 1355-1368 (2015).</li><li>Gaublomme, J. T., Yosef, N., Lee, 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=Single-Cell+Genomics+Unveils+Critical+Regulators+of+Th17+Cell+Pathogenicity.">Single-Cell Genomics Unveils Critical Regulators of Th17 Cell Pathogenicity.</a> Cell. 163 (6), 1400-1412 (2015).</li></ol>

The protocol described herein has been optimized to properly identify T cell subsets present within murine kidneys, aorta and lymph nodes. This protocol can be easily adapted to examine other immune cell subsets such as B lymphocytes and innate immune cells and can be modified to include other tissue types. The digestion step is critical and has to be modified and optimized for each tissue9. A prolonged digestion step or the use of an inappropriate enzyme can affect the stability of antigen expression. Similar...

Evidências recentes demonstram que o sistema imunitário adaptativo, em particular linfócitos T, desempenham um papel crítico no desenvolvimento de várias doenças cardiovasculares 1,2,3,4,5. Por exemplo, no modelo de hipertensão induzida por angiotensina II, uma acumulação de células T nos vasos e nos rins de ratinhos foi descrito 6. A acumulação vascular é predominantemente na adventícia ea gordura perivascular. No rim, as células T acumular tanto na medula e no córtex renal. Dependendo de qual subconjunto está envolvida, estas células T dar origem a diferentes citocinas que podem afectar a função renal e vascular e conduzem ao desenvolvimento da patologia (revisto por McMaster e col. 6). Linfócitos auxiliares T CD4 + pode ser dividido em vários sub-grupos: as células T auxiliares 1 (Th1), Th2, Th9, Th17, Th22, células T reguladoras (Treg), e T auxiliares folicular (TFH) com base nas suas funções e cito assinaturakines 7. Da mesma forma, as células T CD8 + citotóxicas podem ser classificados como Tc1, Tc2, Tc17 ou TC9 8. Existem também células T negativos duplos (isto é, células que não expressam o CD4 ou marcadores de células T CD8 +). Um subconjunto destas células possuem um receptor alternativo gama delta de células T (em vez de os receptores alfa e beta clássicos) e, portanto, são referidas como células T gama delta. A análise de múltiplos parâmetros por citometria de fluxo de marcadores de superfície, citocina e factor de transcrição constitui a melhor abordagem para identificar essas células. Embora este método é amplamente utilizado no campo da imunologia, que é menos bem descritas em órgãos sólidos e na configuração de doenças cardiovasculares. Historicamente, a identificação de linfócitos nos tecidos foi limitado a imuno-histoquímica ou abordagens de RT-PCR. Apesar de imuno-histoquímica e imunofluorescência são poderosos métodos para determinar a distribuição nos tecidos de um antigénio de interest, eles são inadequados para identificar os subconjuntos fenotipicamente envolvidos. Além disso, enquanto que a análise de RT-PCR é útil para detectar a expressão de ARNm de antigénios, citoquinas ou factores de transcrição, que não permite a detecção de proteínas múltiplas simultaneamente ao nível de células individuais. O advento de citometria de fluxo, especialmente quando combinado com a coloração intracelular para detectar citoquinas e factores de transcrição, fornece investigadores com uma técnica poderosa que permite a identificação e quantificação ao nível da célula individual de subconjuntos de células imunitárias em órgãos sólidos. Temos optimizado um ensaio de coloração intracelular para identificar por citometria de fluxo as principais subpopulações de células T presentes no interior do rim murino, da aorta e da aorta nódulos linfáticos de drenagem num modelo de hipertensão induzida por angiotensina II. A optimização de cada passo: digestão do tecido, a activação ex vivo, a permeabilização, e a superfície e os resultados da coloração intracelular em uma altamente reproduzível ensaio que pode ser aplicado a outros modelos de doenças cardiovasculares e renais.

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			<td>10103586001</td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;">Collagenase B</td>
			<td>ROCHE</td>
			<td>11088815001</td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;">Dnase</td>
			<td>ROCHE</td>
			<td>10104159001</td>
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		<tr height="21">
			<td height="21" style="height:21px;">1X Red blood cell lysis buffer</td>
			<td>eBioscience</td>
			<td>00-4333-57</td>
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			<td height="21" style="height:21px;">RPMI Medium 1614 1X</td>
			<td>Gibco</td>
			<td>11835-030</td>
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			<td height="21" style="height:21px;">DPBS without calcium and magnesium</td>
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			<td>14190-144</td>
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			<td>GE Healthcare</td>
			<td>17-5445-02</td>
			<td>For density gradient centrifugation</td>
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			<td height="21" style="height:21px;">GentleMACS &#8482; C tube&#160;</td>
			<td>Miltenyi Biotec</td>
			<td>130-096-334</td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;">GentleMACS dissociator device</td>
			<td>Miltenyi Biotec</td>
			<td>130-093-235</td>
			<td>Use the program SPLEEN_04</td>
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		<tr height="21">
			<td height="21" style="height:21px;">Cell activation cocktail (with Brefeldin A)</td>
			<td>Biolegend</td>
			<td>423303</td>
			<td></td>
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			<td height="21" style="height:21px;">anti-CD16/32</td>
			<td>eBioscience</td>
			<td>14-0161-81</td>
			<td>dilute 1:100</td>
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			<td height="21" style="height:21px;">LIVE/DEAD fixable violet dead cell stain kit</td>
			<td>Life Technologies</td>
			<td>L34955</td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;">Transcription factor buffer set</td>
			<td>BD Pharmingen</td>
			<td>562725</td>
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			<td height="21" style="height:21px;">OneComp eBeads&#160;</td>
			<td>eBioscience</td>
			<td>01-1111-42</td>
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		<tr height="21">
			<td height="21" style="height:21px;">123 count eBeads</td>
			<td>eBioscience</td>
			<td>01-1234-42</td>
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		<tr height="21">
			<td height="21" style="height:21px;">CD45 AmCyan (clone 30-F11)</td>
			<td>BioLegend</td>
			<td>103138</td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;">CD3 PerCP-Cy5.5 (clone 17A2)</td>
			<td>BioLegend</td>
			<td>100218</td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;">IL-17A FITC (clone TC11-18H10.1)</td>
			<td>BioLegend</td>
			<td>506910</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">IL-17F APC (clone 9D3.1C8)</td>
			<td>BioLegend</td>
			<td>517004</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">CD4 APC-Cy7 (clone GK1.5)</td>
			<td>BD Biosciences</td>
			<td>560181</td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;">CD8 APC (clone 53-67)</td>
			<td>eBioscience</td>
			<td>17-0081-82</td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;">T-bet PE-Cy7 (clone 4B10)</td>
			<td>BioLegend</td>
			<td>644823</td>
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		<tr height="21">
			<td height="21" style="height:21px;">IFN&#947; FITC (clone XMG1.2)</td>
			<td>BD Biosciences</td>
			<td>557724</td>
			<td></td>
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intracellular staining and flow cytometry to identify lymphocyte subsets within murine aorta, kidney and lymph nodes in a model of hypertension

It is now well known that T lymphocytes play a critical role in the development of several cardiovascular diseases1,2,3,4,5. For example, studies from our group have shown that hypertension is associated with an excessive accumulation of T cells in the vessels and kidney during the development of experimental hypertension6. Once in these tissues, T cells produce several cytokines that affect both vascular and renal function leading to vasoconstriction and sodium and water retention1,2. To fully understand how T cells cause cardiovascular and renal diseases, it is important to be able to identify and quantify the specific T cell subsets present in these tissues. T cell subsets are defined by a combination of surface markers, the cytokines they secrete, and the transcription factors they express. The complexity of the T cell population makes flow cytometry and intracellular staining an invaluable technique to dissect the phenotypes of the lymphocytes present in tissues. Here, we provide a detailed protocol to identify the surface and intracellular markers (cytokines and transcription factors) in T cells isolated from murine kidney, aorta and aortic draining lymph nodes in a model of angiotensin II induced hypertension. The following steps are described in detail: isolation of the tissues, generation of the single cell suspensions, ex vivo stimulation, fixation, permeabilization and staining. In addition, several fundamental principles of flow cytometric analyses including choosing the proper controls and appropriate gating strategies are discussed.

O protocolo descrito permite a identificação de marcadores de superfície e intracelulares em células T isoladas a partir de rim de murino, da aorta e da aorta nódulos linfáticos de drenagem num modelo de hipertensão induzida por angiotensina II. Os resultados representativos são apresentados abaixo. A Figura 1 mostra a estratégia de propagação usados para identificar a população de células T numa...

Watch this Scientific Journal Video about Intracellular Staining and Flow Cytometry to Identify Lymphocyte Subsets within Murine Aorta, Kidney and Lymph Nodes in a Model of Hypertension at JoVE.com

Intracellular Staining and Flow Cytometry to Identify Lymphocyte Subsets within Murine Aorta, Kidney and Lymph Nodes in a Model of Hypertension

Este artigo fornece metodologia detalhada para identificar e quantificar os subconjuntos de linfócitos T funcionais presentes dentro de rim murino, aorta e linfonodos por coloração intracelular e citometria de fluxo. O modelo de hipertensão induzida por angiotensina II foi escolhido para explicar, passo-a-passo, os procedimentos e os princípios fundamentais da citometria de fluxo e coloração intracelular.

Intracelular Coloração e Citometria de Fluxo para identificar linfócitos subconjuntos dentro Murino Aorta, rim e linfonodos em um modelo de hipertensão

It is now well known that T lymphocytes play a critical role in the development of several cardiovascular diseases1,2,3,4,5. For example, studies from our ...

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