Pregled bibliografske jedinice broj: 888655
Polychromatic flow cytometry in immunophenotyping of primary immunodeficiencies: hidden phenotypes revealed
Polychromatic flow cytometry in immunophenotyping of primary immunodeficiencies: hidden phenotypes revealed // European Academy of Allergy and Clinical Immunology : abstracts
Beč, Austrija, 2016. str. xx-xx (poster, međunarodna recenzija, sažetak, znanstveni)
CROSBI ID: 888655 Za ispravke kontaktirajte CROSBI podršku putem web obrasca
Naslov
Polychromatic flow cytometry in
immunophenotyping of primary
immunodeficiencies: hidden phenotypes revealed
Autori
Polančec, Denis ; Banić, Ivana ; Bulat-Lokas, Sandra ; Živković, Jelena ; Zenić, Lucija ; Turkalj, Mirjana
Vrsta, podvrsta i kategorija rada
Sažeci sa skupova, sažetak, znanstveni
Izvornik
European Academy of Allergy and Clinical Immunology : abstracts
/ - , 2016, Xx-xx
Skup
European Academy of Allergy and Clinical Immunology
Mjesto i datum
Beč, Austrija, 11.06.2016. - 15.06.2016
Vrsta sudjelovanja
Poster
Vrsta recenzije
Međunarodna recenzija
Ključne riječi
children, flow cytometry ; primary immunodeficiencies (PID)
Sažetak
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.
Izvorni jezik
Engleski
Znanstvena područja
Kliničke medicinske znanosti
POVEZANOST RADA
Ustanove:
Dječja bolnica Srebrnjak
Profili:
Sandra Bulat Lokas (autor)
Ivana Banić (autor)
Lucija Zenić (autor)
Denis Polančec (autor)
Mirjana Turkalj (autor)
Jelena Knežević (autor)