Name: Analyse par cytométrie de masse des réponses immunitaires systémiques et locales dans le carcinome hépatocellulaire (Vidéo) | JoVE Journal
Uploaded: 2025-04-25T13:00:00.000+00:00
Duration: 8 min 25 s
Description: Découvrez l'analyse par cytométrie de masse pour profiler les réponses immunitaires systémiques et locales dans le carcinome hépatocellulaire. Cette méthode identifie 14 types cellulaires distincts et révèle les mécanismes d'évasion tumorale pour des thérapies personnalisées.

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5 minutes

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<li>Zhang, Q., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Mass+cytometry-based+peripheral+blood+analysis+as+a+novel+tool+for+early+detection+of+solid+tumours%3A+A+multicentre+study%5BTitle%5D)+AND+%22Gut%22%5BJournal%5D)">Mass cytometry-based peripheral blood analysis as a novel tool for early detection of solid tumours: A multicentre study.</a> Gut. 72 (5), 996-1006 (2023).</li>
<li>Zheng, B., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Trajectory+and+functional+analysis+of+pd-1%28high%29+cd4%28%2B%29cd8%28%2B%29+t+cells+in+hepatocellular+carcinoma+by+single-cell+cytometry+and+transcriptome+sequencing%5BTitle%5D)+AND+%22Adv+Sci%22%5BJournal%5D)">Trajectory and functional analysis of pd-1(high) cd4(+)cd8(+) t cells in hepatocellular carcinoma by single-cell cytometry and transcriptome sequencing.</a> Adv Sci. 7 (13), 2000224(2020).</li>
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<li>Simoni, Y., Chng, M. H. Y., Li, S., Fehlings, M., Newell, E. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Mass+cytometry%3A+A+powerful+tool+for+dissecting+the+immune+landscape%5BTitle%5D)+AND+%22Curr+Opin+Immunol%22%5BJournal%5D)">Mass cytometry: A powerful tool for dissecting the immune landscape.</a> Curr Opin Immunol. 51, 187-196 (2018).</li>
<li>Comi, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Coexpression+of+cd163+and+cd141+identifies+human+circulating+il-10-producing+dendritic+cells+%28dc-10%29%5BTitle%5D)+AND+%22Cell+Mol+Immunol%22%5BJournal%5D)">Coexpression of cd163 and cd141 identifies human circulating il-10-producing dendritic cells (dc-10).</a> Cell Mol Immunol. 17 (1), 95-107 (2020).</li>
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<li>Sheng, J., et al. Human endogenous retrovirus activation contributes to biliary atresia pathogenesis through re-education of resident macrophages. <a target="_blank" href="http://scholar.google.com/scholar?hl=en&safe=off&q=author%3aSheng%2C+J+%22Biorxiv%22">Biorxiv</a>. , (2022).</li>
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<li>Iyer, A., Hamers, A. A. J., Pillai, A. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Cytof+for+the+masses%5BTitle%5D)+AND+%22Front+Immunol%22%5BJournal%5D)">Cytof for the masses.</a> Front Immunol. 13, 815828(2022).</li>
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<li>Zhou, Z., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Deciphering+the+tumor+immune+microenvironment+from+a+multidimensional+omics+perspective%3A+Insight+into+next-generation+car-t+cell+immunotherapy+and+beyond%5BTitle%5D)+AND+%22Mol+Cancer%22%5BJournal%5D)">Deciphering the tumor immune microenvironment from a multidimensional omics perspective: Insight into next-generation car-t cell immunotherapy and beyond.</a> Mol Cancer. 23 (1), 131(2024).</li>
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</ol>

<table><tbody><tr><td>1×PBS</td><td>HyClone</td><td>SH30256.01</td><td></td></tr><tr><td>10×PBS</td><td>HyClone</td><td>SH30256.01</td><td></td></tr><tr><td>100 mm×20 mm tissue-culture-treated culture dish</td><td>Corning</td><td>430167</td><td></td></tr><tr><td>1000 mL pipette tips</td><td>Rainin</td><td>30389218</td><td></td></tr><tr><td>15 mL centrifuge tube</td><td>NEST</td><td>601052</td><td></td></tr><tr><td>200 mL pipette tips</td><td>Rainin</td><td>30389241</td><td></td></tr><tr><td>40 mm nylon cell strainer/70-mm nylon cell strainer</td><td>Falcon</td><td>352340</td><td></td></tr><tr><td>50 mL centrifuge tube</td><td>NEST</td><td>602052</td><td></td></tr><tr><td>70 µm syringe filter</td><td>Sangon Biotech</td><td>F613462-9001</td><td></td></tr><tr><td>Anti-Human CCR2 Antibody (clone: K036C2)</td><td>BioLegend</td><td>357224</td><td></td></tr><tr><td>Anti-Human CCR3 Antibody (clone: 5E8)</td><td>BioLegend</td><td>310724</td><td></td></tr><tr><td>Anti-Human CCR7 Antibody (clone: G043H7)</td><td>BioLegend</td><td>353240</td><td></td></tr><tr><td>Anti-Human CD103 Antibody (clone: Ber-ACT8)</td><td>BioLegend</td><td>350202</td><td></td></tr><tr><td>Anti-Human CD115 Antibody (clone: 9-4D2-1E4)</td><td>BioLegend</td><td>347314</td><td></td></tr><tr><td>Anti-Human CD117 Antibody (clone: 104D2)</td><td>BioLegend</td><td>313201</td><td></td></tr><tr><td>Anti-human CD11b Antibody (clone: 1CRF44)&nbsp;</td><td>BD</td><td>562721</td><td></td></tr><tr><td>Anti-human CD11c Antibody (clone: B-ly6)&nbsp;</td><td>BD</td><td>563026</td><td></td></tr><tr><td>Anti-Human CD123 Antibody (clone: 6H6)</td><td>BioLegend</td><td>306002</td><td></td></tr><tr><td>Anti-Human CD127 Antibody (clone: A019D5)</td><td>BioLegend</td><td>351337</td><td></td></tr><tr><td>Anti-Human CD13 Antibody (clone: WM19)</td><td>BioLegend</td><td>301701</td><td></td></tr><tr><td>Anti-Human CD138 Antibody (clone: MI15)</td><td>BioLegend</td><td>356535</td><td></td></tr><tr><td>Anti-human CD14 Antibody (clone: HCD14)</td><td>BioLegend</td><td>325604</td><td></td></tr><tr><td>Anti-Human CD141 Antibody (clone: M80)</td><td>BioLegend</td><td>344102</td><td></td></tr><tr><td>Anti-Human CD15 Antibody (clone: QA19A61)</td><td>BioLegend</td><td>376302</td><td></td></tr><tr><td>Anti-human CD16 Antibody (clone: B7311)&nbsp;</td><td>BD</td><td>561313</td><td></td></tr><tr><td>Anti-Human CD161 Antibody (clone: HP-3G10)</td><td>BioLegend</td><td>339902</td><td></td></tr><tr><td>Anti-Human CD163 Antibody (clone: GHI/61)</td><td>BioLegend</td><td>333603</td><td></td></tr><tr><td>Anti-Human CD169 Antibody (clone: 7-239)</td><td>BioLegend</td><td>346002</td><td></td></tr><tr><td>Anti-human CD19 Antibody (clone: HIB19)</td><td>BioLegend</td><td>302226</td><td></td></tr><tr><td>Anti-Human CD1c Antibody (clone: L161)</td><td>BioLegend</td><td>331501</td><td></td></tr><tr><td>Anti-Human CD20 Antibody (clone: 2H7)</td><td>BioLegend</td><td>302301</td><td></td></tr><tr><td>Anti-Human CD206 Antibody (clone: 15-2)</td><td>BioLegend</td><td>321151</td><td></td></tr><tr><td>Anti-Human CD24 Antibody (clone: ML5)</td><td>BioLegend</td><td>311129</td><td></td></tr><tr><td>Anti-Human CD25 Antibody (clone: BC96)</td><td>BioLegend</td><td>302624</td><td></td></tr><tr><td>Anti-human CD3 Antibody (clone: UCHT1)&nbsp;</td><td>BD</td><td>555916</td><td></td></tr><tr><td>Anti-Human CD31 Antibody (clone: W18200D)</td><td>BioLegend</td><td>375902</td><td></td></tr><tr><td>Anti-Human CD32 Antibody (clone: FUN-2)</td><td>BioLegend</td><td>303232</td><td></td></tr><tr><td>Anti-Human CD326 Antibody (clone: CO17-1A)</td><td>BioLegend</td><td>369812</td><td></td></tr><tr><td>Anti-Human CD33 Antibody (clone: WM53)</td><td>BioLegend</td><td>303402</td><td></td></tr><tr><td>Anti-human CD4 Antibody (clone: L200)&nbsp;</td><td>BD</td><td>563094</td><td></td></tr><tr><td>Anti-Human CD45 Antibody (clone: HI30)&nbsp;</td><td>BD</td><td>563716</td><td></td></tr><tr><td>Anti-Human CD45RO Antibody (clone: UCHL1)</td><td>BioLegend</td><td>304220</td><td></td></tr><tr><td>Anti-human CD56 Antibody (clone: 5.1H11)</td><td>BioLegend</td><td>362510</td><td></td></tr><tr><td>Anti-Human CD64 Antibody (clone: S18012C)</td><td>BioLegend</td><td>399502</td><td></td></tr><tr><td>Anti-Human CD66b Antibody (clone: 6/40c)</td><td>BioLegend</td><td>392917</td><td></td></tr><tr><td>Anti-human CD68 Antibody (clone: Y1/82A)</td><td>BioLegend</td><td>333808</td><td></td></tr><tr><td>Anti-Human CD69 Antibody (clone: FN50)</td><td>BioLegend</td><td>310902</td><td></td></tr><tr><td>Anti-Human CD7 Antibody (clone: 4H9/CD7)</td><td>BioLegend</td><td>395602</td><td></td></tr><tr><td>Anti-human CD8 Antibody (clone: RPA-T8)&nbsp;</td><td>BD</td><td>557750</td><td></td></tr><tr><td>Anti-Human CD80 Antibody (clone: W17149D)</td><td>BioLegend</td><td>375402</td><td></td></tr><tr><td>Anti-Human CD86 Antibody (clone: BU63)</td><td>BioLegend</td><td>374202</td><td></td></tr><tr><td>Anti-Human FOXP3 Antibody (clone: 206D)</td><td>BioLegend</td><td>320101</td><td></td></tr><tr><td>Anti-Human HLA_ABC Antibody (clone: W6/32)</td><td>BioLegend</td><td>311426</td><td></td></tr><tr><td>Anti-human HLA-DR Antibody (clone: L243)</td><td>BioLegend</td><td>307650</td><td></td></tr><tr><td>Anti-Human IgD Antibody (clone: IA6-2)</td><td>BioLegend</td><td>348211</td><td></td></tr><tr><td>Anti-Human Ki67 Antibody (clone: Ki-67)</td><td>BioLegend</td><td>350501</td><td></td></tr><tr><td>Anti-Human PD_L2 Antibody (clone: MH22B2)</td><td>BD</td><td>567783</td><td></td></tr><tr><td>Anti-Human PD1 Antibody (clone: EH12.2H7)</td><td>BioLegend</td><td>329951</td><td></td></tr><tr><td>Anti-Human PDL1 Antibody (clone: MIH2)</td><td>BioLegend</td><td>393602</td><td></td></tr><tr><td>Anti-human TCR-γδ Antibody (clone: B1)&nbsp;</td><td>BD</td><td>740415</td><td></td></tr><tr><td>Cell cryopreservation solution

analyse par cytométrie de masse des réponses immunitaires systémiques et locales dans le carcinome hépatocellulaire

Mass Cytometry Analysis of Systemic and Local Immune Responses in Hepatocellular Carcinoma

Analyse par cytométrie de masse des réponses immunitaires systémiques et locales dans le carcinome hépatocellulaire

Hepatocellular carcinoma (HCC) is one of the most common and deadliest forms of liver cancer worldwide. Despite advances in treatment, the prognosis for ...

mass-cytometry-analysis-systemic-local-immune-responses

Research

Recherche

JoVE Journal

Biology

Biologie

Nous décrivons ici une méthode pour séparer et enrichir les composants du microenvironnement tumoral immunitaire et non immuns dans les tumeurs sous-cutanées établies. Cette technique permet l’analyse distincte d’infiltrat immunitaire de tumeur et fractions de tumeur non immuns qui peuvent permettre la caractérisation complète du microenvironnement immunitaire de la tumeur.

Enrichissement et la caractérisation des tumeur immunitaire et Non immuns micro-environnements dans les tumeurs murines sous-cutanée établis

Nous décrivons ici une approche protéomique unicellulaires Evaluer immunitaire phénotypique et fonctionnelle (induction de cytokines intracellulaires) des altérations dans les échantillons de sang périphérique, analysés par cytométrie en masse.

Analyse unicellulaires lymphoblastiques et Production de cytokines dans le sang périphérique par cytométrie de flux massique

Ici, nous décrivons le développement d'un modèle de murine médicalement pertinent du cancer du foie récapitulant les dispositifs immunisés typiques du cancer hépatocellulaire (HCC).

Un modèle orthotopic de souris orthopique d'hépatocyteonon oncogène du cancer hépatocellulaire surgissant dans le cadre de l'inflammation et de la fibrose hépatiques

L’évasion de l’éradication des cellules par le tueur naturel (NK) par les cellules cancéreuses est importante pour l’initiation et la progression du cancer. Ici, nous présentons deux protocoles non radioactivité-basés pour évaluer la cytotoxicité cellule-négociée de NK vers les cellules hépatiques de tumeur. En outre, un troisième protocole est présenté pour analyser la migration des cellules NK.

Mesure de la cytotoxicité et de la migration à médiation cellulaire naturelle dans le contexte des cellules tumorales hépatiques

Ici, nous décrivons une stratégie simple et accessible pour visualiser, quantifier, et cartographier les cellules immunitaires dans les sections formelles de tissu tumoral paraffin-intégré. Cette méthodologie combine les techniques d’imagerie et d’analyse numérique existantes dans le but d’élargir la capacité de multiplexage et l’analyse multiparameter des essais d’imagerie.

Visualisation, quantification et cartographie des populations de cellules immunitaires dans le microenvironnement tumoral

Il manque des modèles in vitro complets qui récapitulent fidèlement la maladie humaine pertinente. La présente étude présente la création et la culture de sphéroïdes tumoraux tridimensionnels (3D), un outil in vitro fiable pour étudier l’interaction tumeur-stromale dans le carcinome hépatocellulaire humain.

Un modèle sphéroïde tridimensionnel pour étudier l’interaction tumeur-stromale dans le carcinome hépatocellulaire

Ce protocole décrit la génération de souris du système immunitaire humain (SIH) pour les études d’immuno-oncologie. Les instructions et les considérations relatives à l’utilisation de ce modèle pour tester les immunothérapies humaines sur les tumeurs humaines implantées dans ce modèle sont présentées en mettant l’accent sur la caractérisation de la réponse du système immunitaire humain à la tumeur.

Test d’immunothérapies anticancéreuses dans un modèle murin humanisé portant des tumeurs humaines

Ici, nous détaillons un protocole simple, rapide et fiable assisté par cytométrie en flux multiparamétrique et tumeur sur puce pour surveiller et caractériser les étapes constituant le cycle cancer-immunité. En effet, une compréhension approfondie des interactions entre le cancer et les cellules immunitaires fournit des informations essentielles pour déjouer les tumeurs et guider les soins cliniques.

Co-culture de systèmes in vitro pour reproduire le cycle cancer-immunité

La présence de mastocytes dans la marge interne et les zones péritumorales du carcinome hépatocellulaire après résection confère un pronostic favorable. Cette étude confirme que le logiciel d’analyse d’images QuPath est une plate-forme prometteuse qui pourrait répondre au besoin de reproductibilité, de cohérence et de précision en pathologie numérique.

Les mastocytes dans le microenvironnement du carcinome hépatocellulaire confèrent un pronostic favorable : une étude rétrospective à l’aide du logiciel d’analyse d’images QuPath 

Cette vidéo montre la dissection et l'enlèvement du thymus du fœtus ainsi la préparation des cultures ex vivo de la 2-dGuo traités thymus.

Préparation de la 2-dGuo cultures traitées orgue Thymus

Découvrez l'analyse par cytométrie de masse pour profiler les réponses immunitaires systémiques et locales dans le carcinome hépatocellulaire. Cette méthode identifie 14 types cellulaires distincts et révèle les mécanismes d'évasion tumorale pour des thérapies personnalisées.

Analyse par cytométrie de masse des réponses immunitaires systémiques et locales dans le carcinome hépatocellulaire

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