AUTHOR=Bechi Genzano Camillo , Bezzecchi Eugenia , Carnovale Debora , Mandelli Alessandra , Morotti Elisa , Castorani Valeria , Favalli Valeria , Stabilini Angela , Insalaco Vittoria , Ragogna Francesca , Codazzi Valentina , Scotti Giulia Maria , Del Rosso Stefania , Mazzi Benedetta Allegra , De Pellegrin Maurizio , Giustina Andrea , Piemonti Lorenzo , Bosi Emanuele , Battaglia Manuela , Morelli Marco J. , Bonfanti Riccardo , Petrelli Alessandra TITLE=Combined unsupervised and semi-automated supervised analysis of flow cytometry data reveals cellular fingerprint associated with newly diagnosed pediatric type 1 diabetes JOURNAL=Frontiers in Immunology VOLUME=13 YEAR=2022 URL=https://www.frontiersin.org/journals/immunology/articles/10.3389/fimmu.2022.1026416 DOI=10.3389/fimmu.2022.1026416 ISSN=1664-3224 ABSTRACT=

An unbiased and replicable profiling of type 1 diabetes (T1D)-specific circulating immunome at disease onset has yet to be identified due to experimental and patient selection limitations. Multicolor flow cytometry was performed on whole blood from a pediatric cohort of 107 patients with new-onset T1D, 85 relatives of T1D patients with 0-1 islet autoantibodies (pre-T1D_LR), 58 patients with celiac disease or autoimmune thyroiditis (CD_THY) and 76 healthy controls (HC). Unsupervised clustering of flow cytometry data, validated by a semi-automated gating strategy, confirmed previous findings showing selective increase of naïve CD4 T cells and plasmacytoid DCs, and revealed a decrease in CD56brightNK cells in T1D. Furthermore, a non-selective decrease of CD3+CD56+ regulatory T cells was observed in T1D. The frequency of naïve CD4 T cells at disease onset was associated with partial remission, while it was found unaltered in the pre-symptomatic stages of the disease. Thanks to a broad cohort of pediatric individuals and the implementation of unbiased approaches for the analysis of flow cytometry data, here we determined the circulating immune fingerprint of newly diagnosed pediatric T1D and provide a reference dataset to be exploited for validation or discovery purposes to unravel the pathogenesis of T1D.