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Polychromatic flow cytometry in immunophenotyping of primary immunodeficiencies: hidden phenotypes revealed (CROSBI ID 650930)

Prilog sa skupa u zborniku | sažetak izlaganja sa skupa | međunarodna recenzija

Polančec, Denis ; Banić, Ivana ; Bulat-Lokas, Sandra ; Živković, Jelena ; Zenić, Lucija ; Turkalj, Mirjana Polychromatic flow cytometry in immunophenotyping of primary immunodeficiencies: hidden phenotypes revealed // European Academy of Allergy and Clinical Immunology : abstracts. 2016. str. xx-xx

Podaci o odgovornosti

Polančec, Denis ; Banić, Ivana ; Bulat-Lokas, Sandra ; Živković, Jelena ; Zenić, Lucija ; Turkalj, Mirjana

engleski

Polychromatic flow cytometry in immunophenotyping of primary immunodeficiencies: hidden phenotypes revealed

Research and clinical laboratories addressing investigation and diagnostics of primary immunodeficiencies (PID) use flow cytometry for phenotypical and functional measurements of immune components. Flow cytometry is a powerfull platform for identification and enumeration of various components of the immune system. Multi-laser instruments available today and the development of novel flurochromes conjugated to specific antibodies are used in various combinations. Simoultaneous measurement of up to twenty parameters gives insight into many phenotypically and functionally distinct subsets of leukocytes, any of which might be clinically relevant. The aim of this study was to analyze the phenotype of human peripheral blood lymphocyte subsets in the diagnostics and monitoring of PID using polychromatic flow cytometry assays. Peripheral whole blood samples were stained with antibodies for the cell surface markers: CD45, CD3, CD4, CD8, CD19, CD56 and CD16 as well as for the two activation markers: HLA-DR and CD27. Samples were lysed and fixed with 1x FACSLysing solution (BD Biosciences, USA). Upon centrifugation, leukocyte pellets were resuspended in 1% paraformaldehyde/PBS. Acquisition of the samples was performed using the Navios flow cytometer (Beckman Coulter, USA). The data were analyzed using FlowLogic™ software package (Inivai Technologies, Australia). Peripheral blood from more than 200 patients aged from 2-18 years was analysed in the same manner. A dual-panel platform involving two six color antibody combinations and extensive analysis of the FCS data files resulted in the finding of an increased number of cells with several interesting immunophenotypes: CD19low+CD27high+ B cells, CD3+CD8+CD16high+T cells, CD3-CD16-CD56+CD27+ cells and CD3-CD4+ lymphocytes. Polychromatic flow cytometry implementing an expanded immune monitoring panel in routine immunophenotyping enabled the detection of mononuclear subsets that could not be resolved in protocols where typical four-color immunophenotyping assays were used. In the studies of immune disorders, such as PID, a polychromatic approach using even six colors may allow a deeper insight and understanding of the complex interplay between immune cells and improve diagnostic procedures.

children, flow cytometry ; primary immunodeficiencies (PID)

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Podaci o prilogu

xx-xx.

2016.

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objavljeno

Podaci o matičnoj publikaciji

European Academy of Allergy and Clinical Immunology : abstracts

Podaci o skupu

European Academy of Allergy and Clinical Immunology

poster

11.06.2016-15.06.2016

Beč, Austrija

Povezanost rada

Kliničke medicinske znanosti