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Tytuł pozycji:

Multicentre Harmonisation of a Six-Colour Flow Cytometry Panel for Naïve/Memory T Cell Immunomonitoring

Tytuł:
Multicentre Harmonisation of a Six-Colour Flow Cytometry Panel for Naïve/Memory T Cell Immunomonitoring
Autorzy:
Iole Macchia
Valentina La Sorsa
Irene Ruspantini
Massimo Sanchez
Valentina Tirelli
Maria Carollo
Giorgio Fedele
Pasqualina Leone
Giovanna Schiavoni
Carla Buccione
Paola Rizza
Paola Nisticò
Belinda Palermo
Stefania Morrone
Helena Stabile
Aurelia Rughetti
Marianna Nuti
Ilaria Grazia Zizzari
Cinzia Fionda
Roberta Maggio
Cristina Capuano
Concetta Quintarelli
Matilde Sinibaldi
Chiara Agrati
Rita Casetti
Andrea Rozo Gonzalez
Floriana Iacobone
Angela Gismondi
Filippo Belardelli
Mauro Biffoni
Francesca Urbani
Temat:
Immunologic diseases. Allergy
RC581-607
Źródło:
Journal of Immunology Research, Vol 2020 (2020)
Wydawca:
Hindawi Limited, 2020.
Rok publikacji:
2020
Kolekcja:
LCC:Immunologic diseases. Allergy
Typ dokumentu:
article
Opis pliku:
electronic resource
Język:
English
ISSN:
2314-8861
2314-7156
Relacje:
https://doaj.org/toc/2314-8861; https://doaj.org/toc/2314-7156
DOI:
10.1155/2020/1938704
Dostęp URL:
https://doaj.org/article/04486219890645d4a2e10952819dce83  Link otwiera się w nowym oknie
Numer akcesji:
edsdoj.04486219890645d4a2e10952819dce83
Czasopismo naukowe
Background. Personalised medicine in oncology needs standardised immunological assays. Flow cytometry (FCM) methods represent an essential tool for immunomonitoring, and their harmonisation is crucial to obtain comparable data in multicentre clinical trials. The objective of this study was to design a harmonisation workflow able to address the most effective issues contributing to intra- and interoperator variabilities in a multicentre project. Methods. The Italian National Institute of Health (Istituto Superiore di Sanità, ISS) managed a multiparametric flow cytometric panel harmonisation among thirteen operators belonging to five clinical and research centres of Lazio region (Italy). The panel was based on a backbone mixture of dried antibodies (anti-CD3, anti-CD4, anti-CD8, anti-CD45RA, and anti-CCR7) to detect naïve/memory T cells, recognised as potential prognostic/predictive immunological biomarkers in cancer immunotherapies. The coordinating centre distributed frozen peripheral blood mononuclear cells (PBMCs) and fresh whole blood (WB) samples from healthy donors, reagents, and Standard Operating Procedures (SOPs) to participants who performed experiments by their own equipment, in order to mimic a real-life scenario. Operators returned raw and locally analysed data to ISS for central analysis and statistical elaboration. Results. Harmonised and reproducible results were obtained by sharing experimental set-up and procedures along with centralising data analysis, leading to a reduction of cross-centre variability for naïve/memory subset frequencies particularly in the whole blood setting. Conclusion. Our experimental and analytical working process proved to be suitable for the harmonisation of FCM assays in a multicentre setting, where high-quality data are required to evaluate potential immunological markers, which may contribute to select better therapeutic options.
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