Figure 1.

Serial high-dimensional profiling of INS patients and concomitant immunosuppressive treatments. (A) Study design of thirty children with steroid-dependent nephrotic syndrome in remission with steroids ± calcineurin inhibitors who received either rituximab (RTX) or ofatumumab (OFA) and then underwent withdrawal of immunosuppression in 3 months. After withdrawal, 17 patients developed a relapse, while 13 stayed in remission. Blood was collected before RTX or OFA therapy (T0), after immunosuppressive withdrawal (while still in remission; T1), and at the time of relapse (or at the same time after B cell depleting therapy in patients in remission; T2). Cells were barcoded for patient and time, pooled, and stained with 38 antibodies conjugated to unique metal isotopes. Single-cell data acquired from time-of-flight mass cytometry (CyTOF) was clustered using Phenograph to identify cell clusters and how they evolved over time in each patient (see Methods). (B) Percentage of patients treated with prednisone and (C) average daily prednisone doses in relapsing and non-relapsing patients before anti-CD20 therapy (T0), in remission after immunosuppression withdrawal (T1), and at relapse (or at the same time point after anti-CD-20 therapy in non-relapsing patients; T2). (D) Percentage of patients treated with calcineurin inhibitors (CNI) at the same visits. n=30. *p < 0.05 vs. T0. ^#p < 0.05 vs. relapsing at the same visit. Bar plots depict mean ± SEM.