concentrations  (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: bioRxiv

Article Title: Type I and Type III Interferons Differentially Shape Antiviral Defense and Epithelial Integrity at the Choroid Plexus

doi: 10.64898/2026.02.10.705109

Figure Lengend Snippet: All choroid plexus organoids used were 55±5 days old. (A) Heatmap showing changes in cytokine and chemokine levels in supernatants collected from ChP organoids infected with 200 PFU of E5 at 1, 3, and 5 days post infection, displayed as fold change relative to uninfected controls. (B) Bar plot showing concentrations (log 10 pg/mL) of type III interferons (IFN-λ2/3 and IFN-λ1) and type I interferons (IFN-β and IFN-α2) in supernatants from infected ChP organoids at 1, 3, and 5 days post infection; data are shown as mean ± standard deviation with individual organoids represented as points. (C) Bar plot showing concentrations (log 10 pg/mL) of type III interferons (IFN-λ2/3 and IFN-λ1) and type I interferons (IFN-β) in CSF-like fluid collected from ChP organoids at 2 and 3 days post infection; data are shown as mean ± standard deviation with individual organoids represented as points. (D) Volcano plot showing differential gene expression in ChP organoids at 3 days post infection compared with mock-infected controls. (E) Heatmap showing the top upregulated interferon-stimulated genes (ISGs) in ChP organoids at 3 days post infection. (F) Dot plot from scRNAseq of infected cerebral organoids showing expression of type III IFN receptor subunits ( IFNLR1 and IL10RB ), type I IFN receptor subunits ( IFNAR1 and IFNAR2 ), and selected ISGs ( ISG15 , IFIT2 , and IFITM2 ) across annotated cell clusters; values are scaled such that 1 (red) represents the highest normalized expression and 0 (white) represents the lowest, and dot size indicates the percentage of cells within each cluster expressing the indicated gene. (G–I) ChP organoids were pretreated with 0, 100, or 1,000 ng/mL recombinant human IFN-λ3 prior to infection with 200 PFU of E5. (G) Bar plot showing E5 RNA levels measured by quantitative PCR in ChP organoids following IFN-λ3 pretreatment; data are shown as mean ± standard deviation with individual organoids represented as points. (H) Volcano plot showing differential gene expression in uninfected ChP organoids treated with 1,000 ng/mL IFN-λ3. (I) Heatmap showing the top upregulated ISGs in uninfected ChP organoids treated with 1,000 ng/mL IFN-λ3. For volcano plots, pink circles indicate significantly differentially expressed genes (log₂ fold change > 1; adjusted p value < 0.05). For heatmaps, red indicates higher expression and blue indicates lower expression.

Article Snippet: Choroid plexus organoids (∼55 days in culture) were pretreated overnight with 0, 100, or 1,000 ng/mL recombinant human IFN-λ3 (rhIFN-λ3; R&D Systems, cat. no. 8417-IL-025/CF) diluted in cerebral organoid differentiation medium containing vitamin A.

Techniques: Infection, Standard Deviation, Gene Expression, Expressing, Recombinant, Real-time Polymerase Chain Reaction

Journal: bioRxiv

Article Title: Type I and Type III Interferons Differentially Shape Antiviral Defense and Epithelial Integrity at the Choroid Plexus

doi: 10.64898/2026.02.10.705109

Figure Lengend Snippet: (A-E) Tg32, Tg32 Ifnlr1-/- , Tg32 Ifnar1-/- , and Tg32 Ifnar1-/- Ifnlr1-/- mice were intracranially inoculated with 200 PFU of E5 at postnatal day three, and brains were harvested at two days post infection. (A) Representative hematoxylin and eosin (H&E)–stained sections showing the choroid plexus across all four genotypes; the dotted box indicates the region shown at higher magnification below. (B–D) Blinded pathological scoring of H&E-stained sections for (B) choroid plexus epithelial pathology, (C) immune cell infiltration, and (D) epithelial vacuolation across all four genotypes, with representative images shown below. The scale at right indicates the percentage of tissue affected; red arrows denote representative pathological features. (E) Representative immunohistochemical staining for the tight junction protein ZO-1 in the choroid plexus across all four genotypes; red arrows indicate disrupted junctional staining in Tg32 Ifnar1-/- mice. (F–K) Choroid plexus organoids were pretreated with 0 or 1,000 ng/mL recombinant human IFN-λ3 and subsequently infected with 200 PFU of E5; RNA was collected at three days post infection. (F) Volcano plot showing differentially expressed genes in IFN-λ3–treated choroid plexus organoids, with point colors indicating associated Gene Ontology (GO) pathways (key at right). (G) Heatmap comparing differentially expressed genes across E5-infected, IFN-λ3–treated, and untreated choroid plexus organoids; colors at left denote associated GO pathways (key at right), with red indicating higher expression and blue indicating lower expression. (H–K) Expression changes of selected individual genes across the three conditions. Scale bars: 500 µm and 100 µm (A), 200 µm (B), 100 µm (C), and 50 µm (D, E).

Article Snippet: Choroid plexus organoids (∼55 days in culture) were pretreated overnight with 0, 100, or 1,000 ng/mL recombinant human IFN-λ3 (rhIFN-λ3; R&D Systems, cat. no. 8417-IL-025/CF) diluted in cerebral organoid differentiation medium containing vitamin A.

Techniques: Infection, Staining, Immunohistochemical staining, Recombinant, Expressing