Name: isolation and flow cytometric characterization of murine small intestinal lymphocytes
Uploaded: 2016-05-08T15:00:00
Description: Watch this Scientific Journal Video about Isolation and Flow Cytometric Characterization of Murine Small Intestinal Lymphocytes at JoVE.com

NKS is supported by NIH award K08 AI108690.

Todos os estudos foram conduzidos em análise rigorosa e orientações de acordo com a Comissão dos Assuntos Institucionais animal Cuidado e Uso (IACUC) da Harvard Medical School, que atende aos padrões veterinários estabelecidos pela Associação Americana de Ciência Animal Laboratory (AALAS). 1. Horizontal transmissão de bactérias via co-habitação (Opcional) <ol><li> Para minimizar a contaminação exógena (especialmente se estiver usando camundongos gnotobióticos), praticar uma técnica asséptica...

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W., Brandtzaeg, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tissue+distribution+of+lymphocytes+and+plasma+cells+and+the+role+of+the+gut.">Tissue distribution of lymphocytes and plasma cells and the role of the gut.</a> Trends Immunol. 29 (5), 206-208 (2008).</li><li>Surana, N. K., Kasper, D. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+yin+yang+of+bacterial+polysaccharides:+lessons+learned+from+B.+fragilis+PSA.">The yin yang of bacterial polysaccharides: lessons learned from B. fragilis PSA.</a> Immunol Rev. 245 (1), 13-26 (2012).</li><li>Surana, N. K., Kasper, D. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Deciphering+the+tete-a-tete+between+the+microbiota+and+the+immune+system.">Deciphering the tete-a-tete between the microbiota and the immune system.</a> J Clin Invest. 124 (10), 4197-4203 (2014).</li><li>Gauguet, 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=Intestinal+microbiota+of+mice+influences+resistance+to+Staphylococcus+aureus+pneumonia.">Intestinal microbiota of mice influences resistance to Staphylococcus aureus pneumonia.</a> Infect Immun. , (2015).</li><li>Ochoa-Reparaz, 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=A+polysaccharide+from+the+human+commensal+Bacteroides+fragilis+protects+against+CNS+demyelinating+disease.">A polysaccharide from the human commensal Bacteroides fragilis protects against CNS demyelinating disease.</a> Mucosal Immunol. 3 (5), 487-495 (2010).</li><li>Wu, H. 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=Gut-residing+segmented+filamentous+bacteria+drive+autoimmune+arthritis+via+T+helper+17+cells.">Gut-residing segmented filamentous bacteria drive autoimmune arthritis via T helper 17 cells.</a> Immunity. 32 (6), 815-827 (2010).</li><li>Goodyear, A. W., Kumar, A., Dow, S., Ryan, E. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Optimization+of+murine+small+intestine+leukocyte+isolation+for+global+immune+phenotype+analysis.">Optimization of murine small intestine leukocyte isolation for global immune phenotype analysis.</a> J Immunol Methods. 405, 97-108 (2014).</li><li>Chung, 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=Gut+immune+maturation+depends+on+colonization+with+a+host-specific+microbiota.">Gut immune maturation depends on colonization with a host-specific microbiota.</a> Cell. 149 (7), 1578-1593 (2012).</li><li>Resendiz-Albor, A. A., Esquivel, R., Lopez-Revilla, R., Verdin, L., Moreno-Fierros, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Striking+phenotypic+and+functional+differences+in+lamina+propria+lymphocytes+from+the+large+and+small+intestine+of+mice.">Striking phenotypic and functional differences in lamina propria lymphocytes from the large and small intestine of mice.</a> Life Sci. 76 (24), 2783-2803 (2005).</li><li>Carrasco, 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=Comparison+of+lymphocyte+isolation+methods+for+endoscopic+biopsy+specimens+from+the+colonic+mucosa.">Comparison of lymphocyte isolation methods for endoscopic biopsy specimens from the colonic mucosa.</a> J Immunol Methods. 389 (1-2), 29-37 (2013).</li><li>Van Damme, 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=Chemical+agents+and+enzymes+used+for+the+extraction+of+gut+lymphocytes+influence+flow+cytometric+detection+of+T+cell+surface+markers.">Chemical agents and enzymes used for the extraction of gut lymphocytes influence flow cytometric detection of T cell surface markers.</a> J Immunol Methods. 236 (1-2), 27-35 (2000).</li></ol>

Nós apresentamos um protocolo para o isolamento e caracterização de citometria de fluxo de linfócitos pequenos-intestinal, incluindo a LPLs, IELs e linfócitos no PPs. Para aqueles interessados ​​em avaliar como as mudanças na microbiota afetam o sistema imunológico do intestino delgado, detalhamos os passos simples envolvidos na transmissão horizontal de organismos entre os ratos que abrigam diferentes microbiotas. Embora este protocolo centra-se no intestino delgado, o procedimento é o mesmo para a anális...

A tarefa principal do intestino delgado é muitas vezes considerada a digestão e absorção de nutrientes 1. Enquanto essa função metabólica é claramente essencial, o intestino delgado tem um papel igualmente importante na defesa do hospedeiro, da barragem contínua de antigénios ambientais encontradas no interior do lúmen 2. O tracto intestinal separa o mundo exterior (por exemplo., Antigénios luminais) a partir do ambiente interno do hospedeiro com uma camada epitelial que é apenas uma única camada de células de espessura. Como tal, o sistema imune do intestino delgado tem a tarefa formidável de equilibrar o seu limiar de reactividade, permitindo antigénios estranhos provenientes dos micróbios dieta e comensais de introduzir a mucosa com mínima, se alguma, resposta imune, enquanto a montagem de uma resposta robusta contra patogénios invasores e outras &quot;prejudiciais&quot; antígenos. Respostas imunes excessiva ou inadequada para esses antígenos pode levar a doença patológica (por exemplo., Inflammadoença tory intestino, diabetes tipo I, esclerose múltipla) e deve ser evitado 06/03. No geral, o tracto gastrointestinal é pensado para representar a maior órgão imune no corpo, contendo mais de 70% de todas as células secretoras de anticorpos 7. O sistema imunitário do intestino delgado é constituída por 3 compartimentos principais - a lâmina própria (LP), a camada intraepitelial, e placas de Peyer (PP) - que cada um contém um grupo distinto de linfócitos 2. Os linfócitos LP (LPLs) são principalmente as células TCRαβ + T com ~ células 20% de B; linfócitos intra-epiteliais (IELs) contêm células B muito poucos com mais células TCRγδ + T do que as células TCRαβ + T; e PPS, que são órgãos linfóides secundários embutidos no intestino delgado, parede contêm células ~ 80% de B. Embora cada uma destas regiões anatómicas tem funções ligeiramente distintas e bases ontológicas, elas funcionam em AHmoda armonized para proteger o hospedeiro de insultos patogénicos. Além disso, há um crescente apreciação que a microbiota é um determinante crítico para o desenvolvimento do sistema imunitário intestinal, com o aumento do reconhecimento cognato da relação entre os micróbios e a ontogenia específicos de determinadas linhagens celulares 8,9. Além disso, dado que a formação do sistema imune intestinal afeta as respostas imunes em locais anatomicamente distantes (por ex., Artrite, esclerose múltipla, a pneumonia), tornou-se claro que o desenvolvimento do sistema imunitário do intestino é relevante para mais processos de doença do que anteriormente reconhecido 10 -12. Como tal, o interesse em avaliar quantitativamente o sistema imunológico intestinal foi muito além interações patógeno-hospedeiro para incluir agora interações hospedeiro-comensais e patogênese de muitas doenças sistêmicas bem. Tendo em conta a variabilidade dos métodos actuais naisolamento de linfócitos intestinais, um método que é otimizado para a produção, viabilidade e consistência, equilibrando o tempo necessário é cada vez mais crítica. Os protocolos que envolvem gradientes de Percoll são tempo e trabalho intensivo e potencialmente mais propenso a erro humano, levando a um rendimento variável e viabilidade 13. Aqui, nós fornecemos um protocolo optimizado para o isolamento e caracterização de linfócitos a partir de todos os compartimentos 3 imunes no intestino delgado. Além disso, dado o interesse crescente em alterações induzidas pelo micróbio no sistema imune das mucosas, que incluem passos que podem ser utilizados para permitir a transmissão horizontal de microrganismos entre ratos para avaliar como estas alterações afectam quantitativamente o sistema imune do intestino.

<table>
	<tbody>
		<tr>
			<td>Sterile Gloves</td>
			<td>Kimberly-Clark</td>
			<td>555092</td>
			<td></td>
		</tr>
		<tr>
			<td>sterile mouse cage</td>
			<td>Innovive</td>
			<td>MS2-AD</td>
			<td>contains lid, cage bottom, and alpha-dri bedding</td>
		</tr>
		<tr>
			<td>metal feeder</td>
			<td>Innovive</td>
			<td>M-FEED</td>
			<td></td>
		</tr>
		<tr>
			<td>water bottle</td>
			<td>Innovive</td>
			<td>M-WB-300</td>
			<td></td>
		</tr>
		<tr>
			<td>card holder</td>
			<td>Innovive</td>
			<td>CRD-HLD-H</td>
			<td></td>
		</tr>
		<tr>
			<td>autoclavable rodent chow (NIH-31M)</td>
			<td>Zeigler</td>
			<td>4131207530</td>
			<td></td>
		</tr>
		<tr>
			<td>RPMI medium 1640</td>
			<td>Gibco</td>
			<td>11875-119</td>
			<td></td>
		</tr>
		<tr>
			<td>dithiothreitol (DTT)</td>
			<td>Sigma</td>
			<td>D5545-5G</td>
			<td></td>
		</tr>
		<tr>
			<td>0.5 M EDTA (pH 8.0)</td>
			<td>Ambion</td>
			<td>AM9262</td>
			<td></td>
		</tr>
		<tr>
			<td>fetal bovine serum (FBS)</td>
			<td>GemBio</td>
			<td>100-510</td>
			<td></td>
		</tr>
		<tr>
			<td>dispase II</td>
			<td>Invitrogen</td>
			<td>17105-041</td>
			<td>the concentration in the protocol is based on an activity level of 1.878 U/mg</td>
		</tr>
		<tr>
			<td>collagenase, type II&#160;</td>
			<td>Invitrogen</td>
			<td>17101-015</td>
			<td>the concentration in the protocol is based on an activity level of 245 U/mg</td>
		</tr>
		<tr>
			<td>dissecting scissors</td>
			<td>Roboz</td>
			<td>RS-5882</td>
			<td></td>
		</tr>
		<tr>
			<td>feeding needle (18 G, 2&#34; length)</td>
			<td>Roboz</td>
			<td>FN-7905</td>
			<td></td>
		</tr>
		<tr>
			<td>10 ml syringe</td>
			<td>BD</td>
			<td>305482</td>
			<td></td>
		</tr>
		<tr>
			<td>PBS</td>
			<td>Gibco</td>
			<td>14190-250</td>
			<td></td>
		</tr>
		<tr>
			<td>Disposable Scalpel (15 blade)</td>
			<td>Miltex</td>
			<td>4-415</td>
			<td></td>
		</tr>
		<tr>
			<td>curved forceps</td>
			<td>Roboz</td>
			<td>RS-5211</td>
			<td></td>
		</tr>
		<tr>
			<td>straight forceps</td>
			<td>Roboz</td>
			<td>RS-5132</td>
			<td></td>
		</tr>
		<tr>
			<td>multi-purpose cups, 120 ml</td>
			<td>VWR</td>
			<td>89009-662</td>
			<td></td>
		</tr>
		<tr>
			<td>stir bar</td>
			<td>VWR</td>
			<td>58949-062</td>
			<td></td>
		</tr>
		<tr>
			<td>multi-position stir plate, 9-position</td>
			<td>VWR</td>
			<td>12621-048</td>
			<td></td>
		</tr>
		<tr>
			<td>stainless steel conical strainer, 3 inch&#160;</td>
			<td>RSVP</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>1.5 ml tube</td>
			<td>Eppendorf</td>
			<td>0030 125.150</td>
			<td></td>
		</tr>
		<tr>
			<td>100 &#956;m cell strainer</td>
			<td>Falcon</td>
			<td>08-771-19</td>
			<td></td>
		</tr>
		<tr>
			<td>40 &#956;m cell strainer</td>
			<td>Falcon</td>
			<td>08-771-1</td>
			<td></td>
		</tr>
		<tr>
			<td>50 mL conical tube</td>
			<td>Falcon</td>
			<td>352098</td>
			<td></td>
		</tr>
		<tr>
			<td>1 ml syringe</td>
			<td>BD</td>
			<td>309659</td>
			<td></td>
		</tr>
		<tr>
			<td>96-well plate, round-bottom</td>
			<td>Corning</td>
			<td>3799</td>
			<td></td>
		</tr>
		<tr>
			<td>anti-mouse CD16/32 (Fc block)</td>
			<td>Biolegend</td>
			<td>101320</td>
			<td></td>
		</tr>
		<tr>
			<td>(optional) fixable viability dye eFluor 780</td>
			<td>eBiosciences</td>
			<td>65-0865-18</td>
			<td></td>
		</tr>
		<tr>
			<td>10% formalin, neutral buffered</td>
			<td>Thermo Scientific</td>
			<td>5725</td>
			<td></td>
		</tr>
	</tbody>
</table>

isolation and flow cytometric characterization of murine small intestinal lymphocytes

Os intestinos - que contêm o maior número de células imunes de qualquer órgão do corpo - são constantemente expostos a antígenos estranhos, tanto microbiana e dietética. Dado que uma maior compreensão destes antigénios luminais ajudar a dar forma a resposta imune e que a formação de células imunitárias no intestino é crítica para uma série de doenças sistémicas, tem havido um interesse crescente na caracterização do sistema imune do intestino. No entanto, muitos protocolos publicados são árduos e consumidores de tempo. Apresentamos aqui um protocolo simplificado para o isolamento de linfócitos a partir da própria lâmina do intestino delgado, a camada intraepitelial, e placas de Peyer, que é rápido, reprodutível, e não necessita de gradientes de Percoll laboriosos. Embora o protocolo centra-se no intestino delgado, é também adequado para a análise do cólon. Além disso, destacam-se alguns aspectos que podem precisar de otimização adicional, dependendo do ques científica específicação. Esta abordagem resulta no isolamento de um grande número de linfócitos viáveis ​​que podem, subsequentemente, ser utilizadas para a análise de citometria de fluxo ou meio alternativo de caracterização.

Análise citométrica de fluxo de suspensões de células únicas de linfócitos no intestino delgado deve produzir uma população discreta de células que têm características semelhantes dispersão frontal e lateral como esplenócitos (Figuras 1A e 1B). Os linfócitos podem começam a morrer, se o tecido não é mantida a 4 ° C durante as fases iniciais do isolamento, resultando na população de linfócitos com uma dispersão frontal inferior e sendo mais difíceis...

Watch this Scientific Journal Video about Isolation and Flow Cytometric Characterization of Murine Small Intestinal Lymphocytes at JoVE.com

Isolation and Flow Cytometric Characterization of Murine Small Intestinal Lymphocytes

There is growing interest in the quantitative characterization of intestinal lymphocytes owing to increasing recognition that these cells play a critical role in a variety of intestinal and systemic diseases. In this protocol, we describe how to isolate single cell populations from different small-intestinal compartments for subsequent flow cytometric characterization.

Isolamento e Caracterização de citometria de fluxo de linfócitos de murino Intestino Delgado

The intestines &#8212; which contain the largest number of immune cells of any organ in the body &#8212; are constantly exposed to foreign antigens, both ...

The overall goal of this procedure is to isolate single cell populations from different small intestinal compartments that can subsequently be used for flow cytometric analysis or an alternate means of characterization. This method can help answer key questions in mucosal immunology such as how the microbiota impacts the development of the intestinal immune system. The main advantages of this technique are that they are rapid, reproducible and don't require laborious percoll gradients.To begin this procedure, place the mouse dorsal side down and spray the abdomen with 70 percent ethanol, perform a laparotomy by sequentially cutting the skin and then peritoneal fascia along the ventral midline from the pubic symphysis to the xiphoid process, thus exposing the peritoneal cavity. Next, separate the small intestine from the stomach by transecting the pyloric sphincter. Gently remove the small intestine from the peritoneum and tease away the mesenteric fat.To fully remove the small intestine, make a second cut at the ileocecal junction. Place the isolated small intestine into four degree Celsius RPMI medium containing 10 percent FBS to maximize the cell viability. Gently remove large pieces of fat using curved forceps and use caution to avoid tearing the intestinal tissue itself during the process.To remove the intestinal contents, cannulate the proximal end of the small intestine with an 18 gauge feeding needle affixed to a syringe and gently flush the intestine with 15 to 20 milliliters of cold PBS. Use scissors to excise Peyer's Patches which are located on the antimesenteric side of the small intestine and appear as a multilobulated white mass. Then, place them into cold RPMI medium containing five percent FBS.Next, cut the small intestine into three to four inch segments, remove the residual fat by rolling each small intestinal segment on a paper towel moistened with RPMI and use a dull scalpel to tease the fat away from the tissue. Paying strict attention to the complete removal of mesenteric fat is critical to ensuring the viability of lymphocytes as even minute pieces of fat can cause cells to die. After that, turn the tissue inside out by cannulating the intestinal segments with curved forceps and grasping the distal end of the tissue, then use a pair of straight forceps to gently remove the tissue segment from the proximal end, resulting in the tissue being inverted.Place the tissue segments in a cup containing 30 milliliters of extraction media and a stir bar. Secure the lid on the cap and stir for 15 minutes at 37 degrees Celsius. Stirring should be vigorous, but not turbulent.After 15 minutes, use a steel strainer to separate the tissue pieces from the epithelium containing supernatant which should appear cloudy. Manually agitate the tissue pieces in RPMI medium to wash away the residual extraction media. Then, place the tissue on a dry paper towel, and flip it several times to facilitate the removal of residual mucous that was not liberated by the extraction medium.Subsequently, place the tissue fragments in a one point five milliliter tube with 600 microliters of digestion medium. Mince the tissue until the pieces no longer stick to the scissors and the solution appears homogenous. This step is critical to ensure complete enzymatic digestion of the tissue.Mincing the tissue until the pieces no longer stick to the scissors and the solution appears homogeneous are critical to ensuring cellular yield as well as ensuring reproducibility of results. Add the minced small intestine to a cup containing 25 milliliters of digestion media and stir for 30 minutes at 37 degrees Celsius. Halfway through the digestion, pipe it up and down with a serological pipette to help break down any large chunks of tissue.Next, filter the digestive tissue through a 100 micrometer cell strainer into a 15 milliliter tube. Rinse the strainer with 20 milliliters of RPMI medium containing 10 percent FBS. Afterward, centrifuge the filtered solution for 10 minutes at four degrees Celsius.Carefully decant the supernatant and re-suspend the pellet in one milliliter of RPMI medium containing 10 percent FBS. Filter the re-suspended cells through a 40 micrometer cell strainer into a 15 milliliter tube. Rinse the strainer with 20 milliliters of RPMI containing 10 percent FBS.Subsequently, centrifuge the filtered solution for 10 minutes at four degrees Celsius. Carefully decant the supernatant and re-suspend the pellet in one milliliter of RPMI medium, containing two percent FBS. In this procedure, transfer the excised Peyer's Patches to a cup with 25 milliliters of digestion media and a stir bar.Secure the lid and spin for 10 minutes at 37 degrees Celsius. Afterward, filter the digested Peyer's Patches through a 40 micrometer cell strainer into a 15 milliliter tube. If any clumps remain, press them through the strainer using the flat end of the plunger from a one milliliter syringe.Then, rinse the strainer with 10 milliliters of RPMI medium, containing 10 percent FBS. Centrifuge the filtered solution for 10 minutes at four degrees Celsius. Then, carefully decant the supernatant and re-suspend the pellet in one milliliter of RPMI medium containing two percent FBS.Flow cytometric analysis of single cell suspensions of small intestinal lymphocytes, should yield a discreet population of cells that have similar forward and side scatter characteristics as splenocytes. The lymphocytes may began to die if the tissue is not maintained at four degrees Celsius during the initial stages of the isolation, resulting in the lymphocyte population having a lower forward scatter and being more difficult to separate from other epithelial and dead cells. Moreover, if the mesenteric fat is not completely removed from the small intestinal tissue, there will be a virtually complete loss of lymphocytes.Typically, an 80 percent viability for small intestinal LP lymphocytes is obtained. Once mastered, one can process four mice and have single cell suspensions ready for staining in less than four hours. During this procedure, it's important to completely remove all mesenteric fat and to mince the tissue well to ensure complete digestion.Following this procedure, single cell suspensions can be used for purposes other than flow cytometry, such as en vitro experiments, gene expression analysis or adoptive transfer to further characterize the function of these cells. After its development, this technique paved the way for researchers in mucosal immunology to explore the ontogeny and functional implications of the intestinal immune system in mice. After watching this video, you should have a good understanding of how to isolate single cell suspensions from different small intestinal compartments, by cleaning the mesenteric fat, extracting the epithelial layer, and digesting the tissue with collagenase.

Research

JoVE Journal

Immunology and Infection

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

Isolamento de citometria de fluxo de células primárias Murino Tipo II epiteliais alveolares de Estudos funcionais e Molecular

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

Imunologia e Infecção

Isolation of Lymphocytes from Mouse Genital Tract Mucosa

O isolamento de linfócitos de rato Genital Tract Mucosa

Mucosal surfaces, including in the gastrointestinal, urogenital, and respiratory tracts, provide portals of entry for pathogens, such as viruses and ...

Isolation and Th17 Differentiation of Na&iuml;ve CD4 T Lymphocytes

Isolation and Th17 Differentiation of Na&#239;ve CD4 T Lymphocytes

Isolamento e Th17 Diferenciação de Linfócitos T CD4 Naïve

Th17 cells are a distinct subset of T cells that have been found to produce interleukin 17 (IL-17), and differ in function from the other T cell subsets ...

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

Isolamento e análise de citometria de fluxo de células imunes do cérebro isquêmico Rato

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

Isolation and Flow Cytometric Analysis of Glioma-infiltrating Peripheral Blood Mononuclear Cells

Isolamento e Análise de Citometria de Fluxo infiltrantes Glioma Células Mononucleares do Sangue Periférico

Our laboratory has recently demonstrated that natural killer (NK) cells are capable of eradicating orthotopically implanted mouse GL26 and rat CNS-1 ...

Isolation and Activation of Murine Lymphocytes

Isolamento e Activação de Linfócitos murinos

B and T cells, with their extremely diverse antigen-receptor repertoires, have the ability to mount specific immune responses against almost any invading ...

Isolation and Flow Cytometric Analysis of Human Endocervical Gamma Delta T Cells

Isolamento e Citometria de Fluxo Analysis of Human Endocervical Gama Cells Delta T

The female reproductive tract (FRT) mucosal immune system serves as the first line of defense. Better knowledge of the genital mucosa is therefore ...

Isolating Lymphocytes from the Mouse Small Intestinal Immune System

Isolando os linfócitos de rato pequeno Intestinal sistema imunológico

The intestinal immune system plays an essential role in maintaining the barrier function of the gastrointestinal tract by generating tolerant responses to ...

Isolation and In Vitro Culture of Murine and Human Alveolar Macrophages

Isolation and In Vitro Culture of Murine and Human Alveolar Macrophages

Isolamento e cultura In Vitro de macrófagos alveolares murino e humanos

Alveolar macrophages are terminally differentiated, lung-resident macrophages of prenatal origin. Alveolar macrophages are unique in their long life and ...

Digestion of the Murine Liver for a Flow Cytometric Analysis of Lymphatic Endothelial Cells

Digestão do fígado murino para um fluxo Cytometric análise de células endoteliais linfáticas

Within the liver, lymphatic vessels are found within the portal triad, and their described function is to remove interstitial fluid from the liver to the ...