Usefulness of 6–Colour Multiparameter Flow Cytometry in Canine Lymphoma Phenotyping

Kód výsledku v IS VaVaI

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Výsledek na webu

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DOI - Digital Object Identifier

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Jazyk výsledku

angličtina

Název v původním jazyce

Usefulness of 6–Colour Multiparameter Flow Cytometry in Canine Lymphoma Phenotyping

Popis výsledku v původním jazyce

IV. meeting of the European Canine Lymphoma Network, Lugano, CH, 22.6.2019 - poster. Flow cytometry (FC) is a critical tool in the objective diagnosis and further characterization of lymphoma. Features such as expression of cell surface markers or cell size can provide important prognostic information. In many laboratories, immunophenotyping of canine leukocytes is usually performed using 2-colour or 3-colour FC. The aim of this study was to introduce in praxis a compatible 6–colour multiparameter FC protocol for routine lymphoma phenotypisation using commercially available monoclonal antibodies. For 6-colour FC, directly conjugated (CD3-FITC, CD8-AF700, CD25-eFluor660, CD21-PE) as wells as unconjugated (CD4 and γδTCR) anti-dog monoclonal antibodies and appropriate conjugates (PeCy7 and BV421) in optimal dilution were used. Furthermore, fluorescent minus one controls were prepared for compensation set-up. Fifteen samples of peripheral blood or samples of fine-needle aspirates of peripheral lymph nodes from dogs with suspected lymphoproliferative disease were stained using 2-colour and 6-colour protocol and analysed with a flow cytometer LSRFortessa operated with BD FACSDiva software v. 6.2 (BD Biosciences) to compare the results. From all tested samples, 9 were diagnosed as B-cell and 6 as T-cell lymphoma, incl. 3 T-zonal and 2 γδ-TCR. No spectral overlaps or nonspecific binding of used fluorochromes were detected. Six-colour FC is powerful method to increase the accuracy of lymphoma phenotyping by simultaneous staining of lymphocyte markers with six specific antibodies in one test tube. When compare to 2-colour FC, 6-colour approach proves to be helpful mainly in case of aberrant phenotypes and in small cell types of lymphoma, where neoplastic cells are hardly differentiable from other lymphocytes. Moreover, lesser total amount of samples and reagents are used, since less duplications of the reagents among multiple tubes is required.

Název v anglickém jazyce

Usefulness of 6–Colour Multiparameter Flow Cytometry in Canine Lymphoma Phenotyping

Popis výsledku anglicky

IV. meeting of the European Canine Lymphoma Network, Lugano, CH, 22.6.2019 - poster. Flow cytometry (FC) is a critical tool in the objective diagnosis and further characterization of lymphoma. Features such as expression of cell surface markers or cell size can provide important prognostic information. In many laboratories, immunophenotyping of canine leukocytes is usually performed using 2-colour or 3-colour FC. The aim of this study was to introduce in praxis a compatible 6–colour multiparameter FC protocol for routine lymphoma phenotypisation using commercially available monoclonal antibodies. For 6-colour FC, directly conjugated (CD3-FITC, CD8-AF700, CD25-eFluor660, CD21-PE) as wells as unconjugated (CD4 and γδTCR) anti-dog monoclonal antibodies and appropriate conjugates (PeCy7 and BV421) in optimal dilution were used. Furthermore, fluorescent minus one controls were prepared for compensation set-up. Fifteen samples of peripheral blood or samples of fine-needle aspirates of peripheral lymph nodes from dogs with suspected lymphoproliferative disease were stained using 2-colour and 6-colour protocol and analysed with a flow cytometer LSRFortessa operated with BD FACSDiva software v. 6.2 (BD Biosciences) to compare the results. From all tested samples, 9 were diagnosed as B-cell and 6 as T-cell lymphoma, incl. 3 T-zonal and 2 γδ-TCR. No spectral overlaps or nonspecific binding of used fluorochromes were detected. Six-colour FC is powerful method to increase the accuracy of lymphoma phenotyping by simultaneous staining of lymphocyte markers with six specific antibodies in one test tube. When compare to 2-colour FC, 6-colour approach proves to be helpful mainly in case of aberrant phenotypes and in small cell types of lymphoma, where neoplastic cells are hardly differentiable from other lymphocytes. Moreover, lesser total amount of samples and reagents are used, since less duplications of the reagents among multiple tubes is required.

Druh

O - Ostatní výsledky

CEP obor

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OECD FORD obor

40301 - Veterinary science

Projekt

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Návaznosti

I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Rok uplatnění

2019

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů