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Multicentre harmonisation of a six-colour flow cytometry panel for naïve/memory T cell immunomonitoring
- Source :
- Journal of Immunology Research, Journal of Immunology Research, Vol 2020 (2020)
- Publication Year :
- 2020
- Publisher :
- HINDAWI LTD, 2020.
-
Abstract
- 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.
- Subjects :
- 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
Subjects
Details
- Language :
- English
- Database :
- OpenAIRE
- Journal :
- Journal of Immunology Research, Journal of Immunology Research, Vol 2020 (2020)
- Accession number :
- edsair.doi.dedup.....bdbe1f02cb9e596d34fdb42ef9769b56