Multicentre harmonisation of a six-colour flow cytometry panel for naïve/memory T cell immunomonitoring
| Autor: | Chiara Agrati, Belinda Palermo, Massimo Sanchez, Floriana Iacobone, Mauro Biffoni, Rita Casetti, Giorgio Fedele, Francesca Urbani, Valentina La Sorsa, Pasqualina Leone, Cristina Capuano, Marianna Nuti, Helena Stabile, Iole Macchia, Andrea Rozo Gonzalez, Filippo Belardelli, Carla Buccione, Giovanna Schiavoni, Ilaria Grazia Zizzari, Paola Rizza, Maria Carollo, Cinzia Fionda, Matilde Sinibaldi, Irene Ruspantini, Valentina Tirelli, Concetta Quintarelli, Aurelia Rughetti, Paola Nisticò, Roberta Maggio, Angela Gismondi, Stefania Morrone |
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| Přispěvatelé: | Macchia, I., La Sorsa, V., Ruspantini, I., Sanchez, M., Tirelli, V., Carollo, M., Fedele, G., Leone, P., Schiavoni, G., Buccione, C., Rizza, P., Nistico, P., Palermo, B., Morrone, S., Stabile, H., Rughetti, A., Nuti, M., Zizzari, I. G., Fionda, C., Maggio, R., Capuano, C., Quintarelli, C., Sinibaldi, M., Agrati, C., Casetti, R., Rozo Gonzalez, A., Iacobone, F., Gismondi, A., Belardelli, F., Biffoni, M., Urbani, F. |
| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
CD4-Positive T-Lymphocytes
0301 basic medicine Oncology Receptors CCR7 medicine.medical_specialty CD3 Complex Article Subject Operating procedures Immunology T cells Color CD8-Positive T-Lymphocytes flow cytometry harmonization Immunophenotyping Flow cytometry 03 medical and health sciences 0302 clinical medicine T-Lymphocyte Subsets Internal medicine medicine Humans Immunology and Allergy Observer Variation medicine.diagnostic_test business.industry General Medicine RC581-607 Clinical trial 030104 developmental biology Lazio region medicine.anatomical_structure Italy 030220 oncology & carcinogenesis Leukocytes Mononuclear Leukocyte Common Antigens Immunologic diseases. Allergy business Observer variation Immunologic Memory Immunologic memory Memory T cell Biomarkers Research Article |
| Zdroj: | Journal of Immunology Research Journal of Immunology Research, Vol 2020 (2020) |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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