August 30, 2021
Panyi G., Vmosi G., Bodnr A., Gspr R., Damjanovich S., Looking through ion channels: Recharged concepts in T-cell signaling. obtained by engrafting peripheral blood mononuclear cells (PBMCs) from LN patients into immune-deficient mice. LN mice exhibited features of the disease: increased IFN and CD3+CD8+ T cell renal infiltration, and reduced survival versus healthy donor PBMC engrafted mice. Kv1.3-NP treatment of patient PBMCs before engraftment decreased CD40L/IFN and prolonged survival of LN mice. These data show the potential benefits of targeting Kv1.3 in LN. INTRODUCTION Systemic lupus erythematosus (SLE) is a devastating autoimmune disorder with a wide variety of clinical symptoms predominantly affecting cutaneous, musculoskeletal, cardiovascular, and respiratory C-75 Trans systems. SLE-related complications result in more than 10,000 hospitalizations per year. Lupus IL9 antibody nephritis (LN) occurs in up to 60% of patients with SLE and results in significant mortality and morbidity; 10 to 30% of patients with LN develop C-75 Trans end-stage renal disease requiring dialysis or a kidney transplant (< 0.001), and post hoc testing was performed by Tukeys test, while data in (F) and (G) were analyzed by Students test. Open in a separate window Fig. 2 Immune cell profiling of kidney biopsies from LN, DN, and healthy individuals (NK) with NanoString nCounter Autoimmune Profiling panel.Shown here is the pairwise comparison of the abundance of the total tissue-infiltrating leukocytes (TILs) and the individual immune cell types between for (A) LN (= 4 patients) and NK = 7 individuals) samples and (B) DN (= 7 patients) and NK (= 7 individuals) samples. The abundance of the different immune cell types (at the RNA level) in the kidney biopsies was calculated as log2 cell type scores (see Materials and Methods) and is presented as box and whisker plots. The data are reported as the median (horizontal line), first (top box), and third (bottom box) quartiles, and each symbol represents C-75 Trans a single LN, DN, and NK individual. Statistical significance for the comparative cell type abundance was calculated using two-tailed Students test. The cell scores for a specific cell type can only be compared between two groups (such as NK and LN) but do not support claims that a cell type is more abundant than another cell type within the same group. CD8+ T cells in LN kidneys show increased cytotoxicity and proliferation Studies have shown that infiltration by hyperactive CD8+ C-75 Trans T cells plays a pivotal role in the kidney damage in LN (< 0.001 for (B) to (D)]. Post hoc testing was performed by Dunns test. Open in a separate window Fig. 4 In vitro treatment with Kv1.3-NPs decreases CD40L expression and IFN production in CD45RO+ T cells from patients with LN.(A) Schematic representation of the structure of a lipid NP used to deliver siRNA against Kv1.3 (Kv1.3-NPs) or scramble sequence RNA (scr-NP). PE-PEG-biotin, 1,2-distearoyl-test. In vitro treatment with Kv1.3 NPs decreases CD40L expression and IFN production in Tm cells of patients with LN T cell activation is accompanied by an increase in the cytosolic Ca2+, which activates calcineurin thus inducing NFAT nuclear translocation and downstream transcription of CD40L and inflammatory cytokines, both contributing to the pathogenesis of LN (< 0.001 for all groups). Data in (C) were analyzed by Students test, while data in (D) and (F) were analyzed by one-way ANOVA (< 0.05) and post hoc testing was performed by Holm-Sidak method. Table 1 Immune cell population in LN mice on days 2 and 7 after engraftment.PBMCs from two patients with LN were engrafted in four NSG mice, and immune cell populations were profiled on days 2 and 7 by flow cytometry and are presented as percentages of total live cells. Na?ve T cells were defined as CD3+CD45RO?CD38?FSCintermediate; Tm cells were defined as CD3+CD45RO+CD38?FScintermediate; plasma cells were defined as CD3?CD38+. Data were analyzed by Students test. test, while data in (F) to (H) were analyzed by one-way ANOVA [< 0.001 for (F), < 0.001 for (G), and < 0.001 for (H)] and post hoc testing was performed by Holm-Sidak method. Open in a separate window Fig. 7 CD8+ T cells in the kidneys of LN mice show increased Kv1.3 expression.(A) Representative confocal images of kidney and spleen tissues harvested 6 weeks after engraftment from LN mice that were stained for CD8 (yellow), Kv1.3 (magenta), and nuclei [4,6-diamidino-2-phenylindole (DAPI); cyan]. Scale bar, 50 m. (B) Left: Merged.