Journal: Nature

Article Title: TGFβ links EBV to multisystem inflammatory syndrome in children

doi: 10.1038/s41586-025-08697-6

Figure Lengend Snippet: a-f , Cytokine, chemokine and growth factor profiles of serum from MIS-C patients during the first seven days of hospitalization ( n = 22), untreated samples are marked in red, and at follow-up visits ( n = 28 time points, 23 patients), paediatric patients during an acute infection with a asymptomatic to mild SARS-CoV-2 (mild; n = 57 of which n = 13 had no symptoms) or a moderate to severe (mod.; n = 42 of which teo had severe symptoms), of non-infected children ( n = 40 of which five have an underlying rheumatic condition) and of children six weeks after SARS-CoV-2-infection, that did not develop MIS-C ( n = 11 of which n = 5 have an underlying rheumatic condition). a , Type 1 cytokines, b , type 2 cytokines, c , type 3 cytokines, d , IL-1 family cytokines and e , chemokines and f , growth factors. g , Correlation of cytokine quantity versus time after hospitalization and initiation of treatment. h , Serum levels over time (0 indicates day of initiation of treatment) in individuals with MIS-C are indicated and decline in TGFβ1 serum levels was correlated to time after initiation of treatment. i-j , Graph view results of simple western size-based assay results for i , SMAD2/3 and j , phospho-SMAD2/3 (Ser465/467) (Fig. ) of primary T cells restimulated ex vivo with sera from indicated patients for 30 min with or without addition of a neutralizing anti-TGFβ prior to stimulation. k-m , HEK293T cells were transfected with a plasmid expressing a dominant negative mutant human TGFBR2 or a non-expressing plasmid with Blasticidin-resistance. After positive selection with Blasticidin, cells were stimulated with MIS-C patients’ sera (MIS-C; n = 4; yellow indicates sampling after start of treatment as indicated in i ) for 30 min or left unstimulated, lysed and SMAD2/3 and phospho-SMAD2/3 (Ser465/467) levels were quantified by Simple Western Size-Based assay. k , Phospho-SMAD2/3 (Ser465/467) levels normalised to total SMAD2/3 levels. l-m , Graph view results of simple western size-based assay results for ( l ) SMAD2/3 and m phospho-SMAD2/3 (Ser465/467). n-o , Graph view results of Tubulin used as a loading control for ( n ) T cells and ( o ) transfected HEK293T cells. a-f , Lower limits of quantification are indicated by horizontal lines, short lines indicate medians. A non-parametric ANOVA (Kruskal-Wallis test) was used followed by a Dunn’s multiple comparison test with correction for multiple comparisons, g-h , a two-tailed spearman-correlation was used, or k , a two-tailed ratio paired t-test was used.

Article Snippet: A bead-based multiplex cytokine array (Cytokine/Chemokine/Growth Factor 45-Plex Human ProcartaPlex Panel1, ThermoFisher Scientific) was used according to the manufacturer’s protocol.

Techniques: Infection, Simple Western, Ex Vivo, Transfection, Plasmid Preparation, Expressing, Dominant Negative Mutation, Selection, Sampling, Control, Comparison, Two Tailed Test