The frequency, in terms of CD19+ B cells, of those 10 events was calculated considering the median CD19+ events acquired after RTX treatment, and multiplied by the median absolute B cell count after RTX treatment
The frequency, in terms of CD19+ B cells, of those 10 events was calculated considering the median CD19+ events acquired after RTX treatment, and multiplied by the median absolute B cell count after RTX treatment. (top row plots) and RV-mBc (VLPs-GFP+) (lower row plots) were gated. B. When isotype expression is considered, na?ve B cells and IgM+ mBc express IgD and IgM; switched mBc express IgG and IgA, but a subset which only expresses IgM can also be recognized around the IgD?CD27+ gate: IgM+ only mBc; and CD27- mBc express IgA, IgG or IgM.(TIFF) pone.0097087.s001.tiff (672K) GUID:?C796CF33-DC19-489C-82E5-1D2F8D3A0EF0 Figure S2: RV-memory B cells gating strategies after RTX treatment. A representative result is usually shown for a patient four months after RTX treatment. The gating strategy is the same as the one offered in Physique S1B. The number of acquired events per windows is usually shown in addition to the percentage per subset.(TIFF) pone.0097087.s002.tiff (6.7M) GUID:?02EA6F9B-2CC5-459E-8B13-3393AFAB6427 Physique S3: Comparison of other total B cell subpopulations among the study groups. Other total B cell subpopulations analyzed are shown for healthy volunteers (HV), patients before RTX treatment (prior-RTX) and patients after RTX treatment (after-RTX). The dotted lines represent the estimated flow cytometry detection limit of KX1-004 3.5 CD19+ B cells/mL. Solid lines and error bars denote the median and interquartile range, respectively. Differences between HV (n?=?10) and patients prior-RTX (n?=?10) were evaluated with MannCWhitney assessments and between patients prior-RTX and patients after-RTX (n?=?10) with Wilcoxon assessments. All values reported are 2-tailed.(TIFF) pone.0097087.s003.tiff (520K) GUID:?CA64AFCD-52AF-44D0-9AE7-C982BF93FC0F Physique S4: Comparison of other RV and TT-specific B cell subpopulations among the study groups. Other RV- and TT-specific B cell subpopulations analyzed are shown for HV (n?=?10), patients prior-RTX (n?=?10) and patients after-RTX (n?=?10). Solid lines and error bars denote the median and interquartile range, respectively. Differences between HV and patients prior-RTX were evaluated with MannCWhitney assessments and between patients prior-RTX and patients after-RTX with Wilcoxon assessments. All values reported are 2-tailed.(TIFF) pone.0097087.s004.tiff (516K) GUID:?123DA932-59EF-4C7F-9161-782D6D89BA89 Table S1: Clinical features of patients with autoimmune diseases.(DOCX) pone.0097087.s005.docx (20K) GUID:?7544F132-2CD5-4DB7-A727-96258351424C Abstract The mechanisms that contribute to the KX1-004 maintenance of serological memory are still unclear. Rotavirus (RV) memory B cells (mBc) are enriched in IgM+ and CD27- subpopulations, which are associated with autoimmune diseases pathogenesis. In patients with autoimmune diseases treated with Rituximab (RTX), some autoantibodies (auto-Abs) decrease after treatment, but other auto-Abs and pathogen-specific IgG Abs remain unchanged. Thus, maintenance of autoimmune and pathogen-specific serological memory may depend on the type of antigen and/or Ab isotype evaluated. Antigen-specific mBc and antigen-specific Abs of different isotypes have not been simultaneously assessed in patients after RTX treatment. To study the relationship between mBc subpopulations and serological memory we characterized total, RV- and tetanus toxoid (TT)-specific mBc by circulation cytometry in patients with autoimmune diseases before and after treatment with RTX. We also measured total, RV- and TT-Abs, and some auto-Abs by kinetic nephelometry, ELISA, and EliA assessments, respectively. Minor KX1-004 differences were observed between the relative frequencies of RV-mBc in healthy controls and patients with autoimmune disease. After RTX treatment, na?ve Bc and total, RV- and TT-specific mBc [IgM+, switched (IgA+/IgG+), IgM+ only, IgD+ only, and CD27- (IgA+/IgG+/IgM+)] were significantly diminished. An important decrease Sox18 in total plasma IgM and minor decreases in total IgG and IgA levels were also observed. IgM rheumatoid factor, IgG anti-CCP, and IgG anti-dsDNA were significantly diminished. In contrast, RV-IgA, RV-IgG and RV-IgG1, and TT-IgG titers remained stable. In conclusion, in patients with autoimmunity, serological memory against RV and TT seem to be managed by long-lived plasma cells, unaffected by RTX, KX1-004 and an important proportion of total IgM and serological memory against some auto-antigens seem to be managed by short-lived plasma cells, dependent on mBc precursors depleted by RTX. Introduction Pathogen-specific protective IgG levels following natural contamination or vaccination can persist for decades, or in some cases for a lifetime, in the apparent absence of the antigen [1]. This serological memory provides the host with a first line of defense against reinfection by many microorganisms [2], and crucial pathogen-specific antibody (Ab) titers that correlate with protection have been recognized for several vaccines [3]. Additionally, in autoimmune diseases, autoantibodies (auto-Abs) of different isotypes are associated with disease activity and pathogenesis [4] and in some cases predict disease severity [5]C[7]..