ifnλ2  (R&D Systems)

Journal: Journal of Human Immunity

Article Title: Increased prevalence of autoantibodies neutralizing IFNλ2/3 in young individuals with cystic fibrosis

doi: 10.70962/jhi.20250268

Figure Lengend Snippet: At least 10% of young individuals with CF harbor neutralizing anti-IFNλ2/3 IgG autoAbs. (A) IFNλ1/2/3 neutralization by plasmas from individuals with AD ( n = 39), various adults (CT; n = 68), or pwCF (CF; n = 84 samples from n = 51 individuals). Dotted lines indicate thresholds set using standard outlier calculations. Neutralizing plasmas are orange; numbers represent unique individuals; positive individuals are labeled. (B) Anti-IFN-III and anti-IFN-I IgGs in CF plasmas positive (neut; n = 9) or negative (non-neut; n = 6) for IFNλ2/3 neutralization. (C) Spearman’s correlation between IFNλ2/3 neutralization (A) and IgG (B). (D) Anti-IFNλ2/3 IgG subclasses in CF plasmas ( n = 5). (E) Area under the curve from PIV5-GFP replication kinetics in A549 cells pretreated with IFNλ2 (or not) in the presence of pwCF plasmas positive (pos) or negative (neg) for IFNλ2/3 neutralization. Values are triplicate means from two experiments. Error bars indicate standard deviations. For the IFNλ2+pos and IFNλ2+neg groups, data are from two patients (squares/triangles) from two experiments. (F) Fluorescence images of infected cells from E. (G) Random forest regression of clinical variables in pwCF positive ( n = 4) or negative ( n = 16) for anti-IFNλ2/3 autoAbs. Dots represent importance scores per iteration (maximum 5,000); error bars reflect importance spread. Abbreviations: MFI FC, median fluorescence intensity fold-change; IVIG, intravenous immunoglobulin; IBD, inflammatory bowel disease; CFRD, CF-related diabetes; ABPA, allergic bronchopulmonary aspergillosis; SA, Staphylococcus aureus ; PA, Pseudomonas aeruginosa . Statistical analyses were two-way ANOVA with the Šidák correction (B), Spearman correlation (C), and Mann–Whitney test (E) (ns, nonsignificant; *P < 0.05; ****P < 0.0001).

Article Snippet: Replication of GFP-encoding PIV5 (PIV5-GFP; P523; ViraTree) in A549 cells was assayed as described previously , assessing the effects of 1:100 diluted plasma samples and 10 ng/ml IFNλ2 (8417-IL; R&D Systems).

Techniques: Neutralization, Labeling, Fluorescence, Infection, MANN-WHITNEY

Journal: Frontiers in Immunology

Article Title: Iron improves the antiviral activity of NK cells

doi: 10.3389/fimmu.2024.1526197

Figure Lengend Snippet: Increased activation and transferrin receptor expression after IFNα stimulation of NK cells. Expanded NK cells were stimulated with IFNα11 (500 U/ml), IFNβ (500 U/ml), IFNλ2 (100 ng/ml), IL-2/12 (20 ng/ml, 10 ng/ml) for 18 hours or left unstimulated (unstim.). NK cells (viable, CD3 - , NK1.1 + ) were analyzed for activation (CD69) (A) . Representative dot plots and bar graphs of CD71 + NK cells and transferrin receptor (CD71) expression on CD71 + NK cells are shown in (B–D) . Statistically significant differences between the groups were analyzed with an Ordinary one-way ANOVA. All data are presented as mean values ± SEM. In (E, F) , correlation of CD71 (MFI, %) and CD69 (%) is shown. Splenocytes were stimulated or left untreated overnight and the uptake of transferrin was measured in NK cells by flow cytometry (G) . Correlation of transferrin uptake and activation is shown in (H) . Five mice from 3 independent experiments were used and analyzed by an ordinary one-way ANOVA. Black circles represent unstimulated, open circles IFNα-, grey IFNβ-, dark grey IFNλ- and bright grey IL-2/-12-stimulated NK cells. In (I) , the NK cell proliferation was analyzed by using KI-67. For (A, C–F) five mice per group collected from five independent experiments were used (n=5).

Article Snippet: On day 6, IL-2 (20 ng/ml), IL-12 (10 ng/ml), murine IFNα11 (500 U/ml) , murine IFNβ (500 U/ml) or murine IFNλ2 (100 ng/ml, Peprotech) were added for 18 hours, if indicated.

Techniques: Activation Assay, Expressing, Flow Cytometry