Combined unsupervised and semi-automated supervised analysis of flow cytometry data reveals cellular fingerprint associated with newly diagnosed pediatric type 1 diabetes

• DOI: 10.3389/fimmu.2022.1026416
• Publication Date: 2022-10-27T06:33:49Z
• Authors: Camillo Bechi Gen...; Eugenia Bezzecchi; Debora Carnovale; Alessandra Mandelli; Elisa Morotti; Valeria Castorani; Valeria Favalli; Angela Stabilini; Vittoria Insalaco; Francesca Ragogna; Valentina Codazzi; Giulia Maria Scotti; Stefania Del Rosso; Benedetta Allegra...; Maurizio De Pelle...; Andrea Giustina; Lorenzo Piemonti; Emanuele Bosi; Manuela Battaglia; Marco J. Morelli; Riccardo Bonfanti; Alessandra Petrelli
• Keywords: type 1 diabetes; flow cytometry; Immunology; computational biology; fingerprints; flow cytometry; immune markers; pediatric diabetes; type 1 diabetes;; fingerprints; RC581-607; Flow Cytometry; T-Lymphocytes, Regulatory; 3. Good health; Killer Cells, Natural; 03 medical and health sciences; computational biology; Diabetes Mellitus, Type 1; 0302 clinical medicine; immune markers; Humans; Immunologic diseases. Allergy; Child; pediatric diabetes; Autoantibodies

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.