Introduction:We have reported a predominant Th2 immune response in steroid-dependent nephrotic syndrome (SDNS).This study aimed to study ILC2s contribution to the Th2-biased phenotype in idiopathic nephrotic syndrome (INS) patients in relapse as well as recapitulate INS through adoptive cell transfer of patient-specific ILC2s in immunocompromised mice.Methods: A total of 14 patients with childhood-onset INS (relapse and remission paired samples) and 7 age-matched healthy controls were recruited.ILC2s were isolated through removal (positive selection) of CD19, CD4, CD8, CD14 followed by ILC2s negative selection (Miltenyi).To investigate ILC2-induced proteinuria in vivo, ILC2s from SDNS patients in relapse were adoptive transferred to NOD scid-gamma (NSG) mice.For in vitro study, ILC2 cells were cultured for 72 hours with IL-33, IL-25, TSLP (10ng/ml) and IL-2 (20ng/ml) and the culture supernatants were collected for cytokine measurement.GATA3 expression levels were examined in ILC2s though incubation of PBMC with IL-33 (10ng/ml) for 72h, with or without Dexamethasone (Dex) (0.1mol/ml).ILC subsets and GATA3 were analysed using flow cytometry, with ILC2s defined as CD45+Lin-CD127+CRTH2+.Statistical analysis was done using Mann-Whitney U tests and Wilcoxon signed-rank test for paired samples.Results: INS patients in relapse had higher ILC2s levels compared to controls (19.02.2%vs 9.62.3%,P=0.01) with concomitant elevated production of Th2 and Th2-related cytokines (IL-5, IL-13, IL-9, IL-10 and Eotaxin (P0.99), in keeping with Th2 polarisation in steroid-dependent disease.Following remission, ILC2s levels were significantly decreased (P=0.01,paired).Dexamethasone incubation of PBMC in relapsed SDNS patients resulted in downregulation of ILC2 GATA3 levels (mean difference 13.96%3.21%,P=0.02, paired), but paradoxically increased ILC2 cell numbers (mean difference 5.81.1%,P<0.001).
Inoue et al. (Wed,) studied this question.