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28.09.2018 | Brief Report | Ausgabe 1/2019

Pediatric Nephrology 1/2019

B cell phenotype in pediatric idiopathic nephrotic syndrome

Zeitschrift:
Pediatric Nephrology > Ausgabe 1/2019
Autoren:
Manuela Colucci, Rita Carsetti, Simona Cascioli, Jessica Serafinelli, Francesco Emma, Marina Vivarelli
Wichtige Hinweise

Electronic supplementary material

The online version of this article (https://​doi.​org/​10.​1007/​s00467-018-4095-z) contains supplementary material, which is available to authorized users.

Abstract

Background

A pathogenic role of B cells in non-genetic nephrotic syndrome has been suggested by the efficacy of rituximab, a B cell depleting antibody, in maintaining a prolonged remission. However, little information is available on B cell homeostasis in nephrotic syndrome patients.

Methods

We retrospectively analyzed by flow cytometry the distribution of different B cell subpopulations in 107 steroid-sensitive and in 6 genetic steroid-resistant nephrotic syndrome pediatric patients, compared with age- and sex-matched controls.

Results

Fifty-one steroid-sensitive patients at disease onset, before starting immunosuppression, presented significantly increased levels of total, transitional, memory, and switched memory B cells compared to controls. Oral immunosuppression strongly affected transitional and mature B cell levels in 27 patients in relapse and also in 29 patients in remission, whereas memory B cells were significantly higher compared to controls during relapse, despite the immunosuppressive treatment, and were normalized only in patients in remission. Children with genetic forms of steroid-resistant nephrotic syndrome presented no differences in B cell profile from controls.

Conclusions

Our study indicates that memory B cells, more than other B cell subsets, are increased and appear to be pathogenically relevant in steroid-sensitive nephrotic syndrome pediatric patients.

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Zusatzmaterial
Supplementary Table 1 (DOCX 20 kb)
467_2018_4095_MOESM1_ESM.docx
Supplementary Figure 1. Gating strategy to discriminate the different B cell subpopulations by multicolor flow cytometry analysis. (JPG 766 kb)
467_2018_4095_MOESM2_ESM.jpg
Supplementary Figure 2. Analysis of circulating B cell subpopulations in pediatric steroid-sensitive nephrotic syndrome. Levels of B-cell subsets, expressed as cells/μl of blood, from patients at disease onset (n=51), in relapse (n=27) or in remission (n=29). (A) Gated total CD19+ B cells were identified based on the expression of surface markers as depicted in Fig. S1: (B) transitional, (C) mature, (D) memory, (E) IgM memory, and (F) switched memory B cells. Each box plot represents the median and the 25th and 75th centiles. Error bars represent the smallest and the largest values. Differences between groups were compared using the nonparametric Kruskal-Wallis test and, if significant, unpaired Mann-Whitney U test was applied. *, p<0.05; **, p<0.01; ***, p<0.001. (JPG 485kb)
Supplementary Figure 3. Multicolor flow cytometry analysis of circulating B-cell subpopulations in children with genetic forms of steroid-resistant nephrotic syndrome. Levels of B-cell subsets from patients with genetic forms of steroid-resistant nephrotic syndrome (n=6) were compared with values of the same number of age- and sex-matched healthy donors (HD). (A) Gated total CD19+ B cells were identified on the basis of the expression of surface markers as depicted in Fig. S1; (B) transitional, (C) mature, (D) total memory, (E) IgM memory, and (F) switched memory B cells were expressed as percentages of total lymphocytes. Each box plot represents the median and the 25th and 75th centiles. Error bars represent the smallest and the largest values. Differences between groups were compared using the nonparametric unpaired Mann-Whitney U test. (JPG 572kb)
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