Journal: Cell Reports Medicine

Article Title: Rewired type I IFN signaling is linked to age-dependent differences in COVID-19

doi: 10.1016/j.xcrm.2025.102285

Figure Lengend Snippet: Gradual age-dependent change in the phenotype of infection-induced CD4 + T cells and B cells (A) UMAP, showing pre-gated CD4 + T cells from the CyTOF dataset. 19 clusters have been produced using a semi-supervised approach with FlowSOM algorithm. UMAP presents all the patients that were part of the dataset and used for clustering, including follow-up measurements of some patients done approximately 2 weeks and 6 months after the first, acute infection phase measurement (details in ). (B) UMAPs, showing the location of cells belonging to the respective group (colored in, whereas gray identifies cells in other groups). Gray outlines indicate cluster regions enriched in infected children or adults. (C) Boxplots, showing relative abundance of infection-induced clusters resulting from the FlowSOM algorithm, calculated per sample within all CD4 + T cells from the CyTOF data. Kruskal-Wallis + Wilcoxon p value. (D) Scatterplots, showing mean Z score normalized CD38 and CCR6 expression in relationship to patient age for infected patients (linear model fitted to data and Spearman’s rank correlation coefficient in black) within CD4 + T cell CyTOF data. (E) UMAP, showing pre-gated B cells from the CyTOF dataset. 15 clusters have been produced using a semi-supervised approach with FlowSOM algorithm. (F) UMAPs, showing the location of cells belonging to the respective group (colored in, whereas gray identifies cells in other groups). Gray outlines indicate cluster regions enriched in infected children or adults. (G) Boxplots, showing relative abundance of infection-induced clusters resulting from the FlowSOM algorithm, calculated per sample within all B cells from the CyTOF data. Kruskal-Wallis + Wilcoxon p values. (H) Scatterplots, showing mean Z score normalized CXCR5 and CD69 expression in relationship to patient age for infected patients (linear model fitted to data and Spearman’s rank correlation coefficient in black) within B cell CyTOF data. (I) Time-dependent stacked line graphs, displaying the relative mean cluster abundance of all activated CD4 + T cell clusters determined by CyTOF. Activated clusters were defined as clusters having above-average Z -scored expression of activation markers (CD25, HLA-DR, CD38, CD137, CD69, and Ki67) compared to other clusters. Patient group color-coded figurines on the right-hand side point out cluster accumulation patterns. (J) Scatterplot, showing the sum of the relative abundance of all activated CD4 + T cell clusters determined by CyTOF.

Article Snippet: CD38 Inh_78c , Tocris, Biotechne , 6391.

Techniques: Infection, Produced, Expressing, Activation Assay

Journal: Cell Reports Medicine

Article Title: Rewired type I IFN signaling is linked to age-dependent differences in COVID-19

doi: 10.1016/j.xcrm.2025.102285

Figure Lengend Snippet: Consequences for local T cell responses and generated antibody profiles (A) Dotplot, showing scaled average expression of genes in TCRab + T cells, subset from the nasal swab scRNA-seq data. Clusters, increased with infection (0, 4, 5, 6, and 7). A total of 8 clusters have been produced using a graph-based approach as implemented in Seurat package (KNN graph with Louvain community detection). A horizontal line splits the dotplot in two parts; genes above the line were curated based on the presence of clusters with pronounced ISG signature and include other genes useful for annotation; genes below the line were found to be differentially expressed between the clusters (FindMarkers Seurat function). (B) Scatterplots showing CD38 and TNF genes transcription (average scaled expression in clusters 0, 4, 5, 6, and 7, expanded with infection) for each donor, plotted against donor’s age, using TCRab + T cells, subset from nasal swab scRNA-seq data. Linear models fitted to the data points and Spearman’s rank correlation coefficients. (C) Stacked bar chart showing relative expression strength of heavy-chain genes encoding for the different IgG and IgA isotypes in plasmablasts (B cell cluster 12, PBMC scRNA-seq experiment). Plasmablasts, expressing either of the heavy-chain genes, were pre-selected. Expression values for each gene were calculated and normalized to the total expression of all heavy-chain genes. (D) Boxplots of S1-specific IgG (left) and IgA (right) antibody titers for the acute infection phase and follow-up measurements done approximately 2 weeks and 6 months later. Titers for second and third time points are normalized to the first time point for each patient (fold change and ratio). Wilcoxon p values.

Article Snippet: CD38 Inh_78c , Tocris, Biotechne , 6391.

Techniques: Generated, Expressing, Infection, Produced

Journal: Cell Reports Medicine

Article Title: Rewired type I IFN signaling is linked to age-dependent differences in COVID-19

doi: 10.1016/j.xcrm.2025.102285

Figure Lengend Snippet: Mechanistic in vitro studies link age-dependent rewiring of type I IFN responsiveness with in vivo -detected opposite activation profiles (A) Overview of the workflow used to study the responsiveness to type I IFN and IL-1b. PBMCs from uninfected children and adults were stimulated with either SEB or a combination of SEB, ODN CpG2216, B18R, and recombinant IFNa. In a parallel experiment series, different concentrations of IFNa as well as combinations of SEB, ODN CpG2216, IL-1b, and IL-1b inhibitor anakinra were tested. After 4 days of incubation, phenotypic differences in activation marker expression were determined by flow cytometry, while cell culture supernatants were used for cytokine and chemokine quantification. Experiments focused on IL-1b and anakinra influence were only measured in cytokine proteomics. (B) Boxplot of arcsinh-transformed median CD38 fluorescence intensity in proliferating CD4 + T cells, showing influence of CpG2216-mediated activation on CD38 expression. Wilcoxon test p values. Dropout in uninfected children group SEB condition is due to low cell number. (C) Boxplot of arcsinh-transformed median CD38 fluorescence intensity in proliferating CD4 + T cells, showing influence of CpG2216-mediated activation and IFNa (30 ng/mL) on CD38 expression in children. Wilcoxon test p values. Dropouts in SEB and SEB+IFNa perturbations are due to low cell number. (D) Boxplots of CD38 median signal intensity in proliferating CD4 + T cells, separated into CD45RA − (violet filling) memory and CD45RA + naive subpopulations, showing the difference in CD38 upregulation in response to CpG2216-mediated activation and IFNa release between memory and naive CD4 + T cells. Wilcoxon test p values. Dropout in uninfected children, SEB perturbation is due to low cell number. (E) Boxplot of IFNa concentration measured in cell culture supernatant and normalized to values detected in SEB condition for each patient, showing the effectiveness of CpG2216 in provoking IFNa release as well as of B18R in reducing the concentration of soluble IFNa. Dropout in uninfected children is due to low cell number. (F) Heatmap, showing scaled average log concentration of the 18 cytokines measured in co-culturing experiments for different perturbations using PBMC. (G) Line plots, showing the dependence of IFNg, IL-21, and IL-1b concentrations on the IFNa concentration. Wilcoxon p values. (H) Scatterplot, illustrating the correlation between donor age and IL-1b concentration in supernatant when PBMCs are stimulated with SEB and CpG.

Article Snippet: CD38 Inh_78c , Tocris, Biotechne , 6391.

Techniques: In Vitro, In Vivo, Activation Assay, Recombinant, Incubation, Marker, Expressing, Flow Cytometry, Cell Culture, Transformation Assay, Fluorescence, Concentration Assay

Journal: Journal of nanobiotechnology

Article Title: Bionic bearing-inspired lubricating microspheres with Immunomodulatory effects for osteoarthritis therapy.

doi: 10.1186/s12951-025-03544-2

Figure Lengend Snippet: Fig. 1 Characterization of 78c@Lipo-FA. (A) Scheme diagram of the preparation process of Lipo-FA; materials including 78c, HPSC, cholesterol, octadecy lamine, and DSPE-PEG2k-FA. (B) TEM of liposomes. (C) Particle size of liposomes (n = 3). (D) Zeta potential of liposomes (n = 3). (E) Representative panels of the uptaking-liposomes in the BMMs assayed by flow cytometry. (F) Representative images of the uptaking-liposomes in the BMMs observed by the fluorescence microscope (After BMMs were incubated for 4 h). (G) Relative fluorescence intensity of the uptake of liposomes by BMMs (n = 3). ***p < 0.001

Article Snippet: The CD38 inhibitor (78c) was purchased from Selleck (Wuhan, China).

Techniques: Liposomes, Zeta Potential Analyzer, Flow Cytometry, Fluorescence, Microscopy, Incubation

Journal: Clinical & Translational Immunology

Article Title: Targeting CD38 with monoclonal antibodies disrupts key survival pathways in paediatric Burkitt's lymphoma malignant B cells

doi: 10.1002/cti2.70011

Figure Lengend Snippet: Correlation analysis of CD38 and MYC gene expression in pBL patient samples. (a) Unsupervised PCA analysis shows the cluster pattern of the 29 paediatric lymphoma samples (GSE10172) based on significantly differentially expressed genes among these samples (1800 genes, one‐way ANOVA with the post hoc Tukey test P = 0.01); the colour key for disease subtypes in this panel is consistent with those used in the following panels of this figure. (b) PCA chart shows the distribution of the significantly differentially expressed genes that lead to the samples clustering in a . (c) Heatmap shows the 1800 significantly differentially expressed genes and the hierarchical distribution of the samples reflecting the clustering in a . The disease subtype and absence/presence of the IgH‐MYC translocation related to these samples are also reported. Correlation analysis for MYC and CD38 gene expression was performed on the samples contained in the dataset GSE10172 and Pearson's correlation coefficient ( r ), coefficient of determination ( R 2 ) and P ‐value were calculated for (d) the whole 29 samples contained in the data set, (e) only for BL and BL‐like samples, (h) only for BL samples. (g) Same analysis as in (d–f) but on 11 pBL samples contained in the GSE64905 dataset. (h) Same analysis as in (d–g) but on 19 pBL samples contained in GSE10172 and GSE64905 data sets. Here, the data sets were merged and normalised using the Z ‐score normalisation method.

Article Snippet: The CD38 cyclase assay was conducted using the CD38 Inhibitor Screening Assay Kit (BPS Bioscience, San Diego, California, USA) according to the manufacturer's protocol.

Techniques: Expressing, Translocation Assay

Journal: Clinical & Translational Immunology

Article Title: Targeting CD38 with monoclonal antibodies disrupts key survival pathways in paediatric Burkitt's lymphoma malignant B cells

doi: 10.1002/cti2.70011

Figure Lengend Snippet: DARA and ISA differential interaction with CD38's structure and its cyclase activity and impact on pBL cell proliferation, apoptosis and cell cycle. (a, b) The extracellular domain's structure of CD38 from two perspectives (top and 180° rotated bottom view, respectively), highlighting the epitopes recognised by DARA and ISA and the active site's location. (c) The crystal structure of the unbound CD38 ectodomain (PDB entry: 1YH3), showing the accessible active site. Arrows indicate the predicted binding sites for DARA and ISA. The dashed box indicates active site. A zoomed‐in view of the active site is provided in the adjacent box. (d) The crystal structure of CD38's ectodomain complexed with NAD + (upper panel) and ADPR (lower panel) in its active site (PDB entries: 3OFS and 8P8C respectively). The dashed box indicates active site, NAD + and ADPR. A zoomed‐in view of the active site is provided in the adjacent box. (e, f) The conformational changes of CD38 when bound to the Fab region of DARA (PDB entry: 7DHA) and ISA (PDB entry: 4CMH), respectively. Dashed boxes indicate the active site. A zoomed‐in view of the active site is provided in the adjacent boxes. (g) Concentration‐dependent inhibition of CD38 cyclase activity by DARA, ISA and quercetin, a CD38 cyclase inhibitor. The assay was performed on recombinant CD38 protein, with untreated controls (represented by the blue dotted line, No inh.) set as 100%. (h, i) The cell number and % of dead cells (7AAD + ) analysis in Ramos cells after a 4‐day culture period, comparing untreated (−) to treated with DARA, ISA or beriglobin control. (j) Cell cycle stages in Ramos cells stained with Vibrant DyeCycle in the same experimental setup as (h, i) . (k) % of cells in G1, S and G2/M phase. Statistical significance was calculated with one‐way ANOVA with the post hoc Tukey test and denoted as * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001. Data are presented as mean ± SD. Non‐significance is not indicated in the figure. Data in (g) are from two independent experiments with n = 3 replicates each, with results normalised and combined. Data in (h–k) are representative of one experiment with n = 3 replicates. Independent experiments were repeated at least twice.

Article Snippet: The CD38 cyclase assay was conducted using the CD38 Inhibitor Screening Assay Kit (BPS Bioscience, San Diego, California, USA) according to the manufacturer's protocol.

Techniques: Activity Assay, Binding Assay, Concentration Assay, Inhibition, Recombinant, Control, Staining

Journal: Clinical & Translational Immunology

Article Title: Targeting CD38 with monoclonal antibodies disrupts key survival pathways in paediatric Burkitt's lymphoma malignant B cells

doi: 10.1002/cti2.70011

Figure Lengend Snippet: Comparative efficacy of DARA and ISA on modulating IgM:CD19 and IgD:CD19 interactions. (a) Fab‐PLA study of the proximity of IgM to CD19 on Ramos cells unstimulated or 5‐min anti‐IgM–stimulated without and with exposure to DARA or ISA (top to bottom). PLA signals are shown in red and nuclei in blue. Scale bar, 5 μm. (b) Scatter dot plot represents the mean of PLA signals for IgM:CD19 interaction (signal counts). (c) Fab‐PLA study of the proximity of IgD to CD19 on Ramos cells unstimulated or 5‐min anti‐IgM–stimulated without and with exposure to DARA or ISA (top to bottom). PLA signals are shown in red and nuclei in blue. Scale bar, 5 μm. (d) Scatter dot plot represents the mean of PLA signals for IgD:CD19 interaction (signal counts). Shown are representative microscope images (a, c) . Statistical significance in this figure was calculated with one‐way ANOVA with the post hoc Tukey test and denoted as * P < 0.05; ** P < 0.01; **** P < 0.0001. Non‐significance is not indicated in the figure. Independent experiments were repeated twice. In these graphs, every data point is one cell; error bars show mean ± SD. (e) Schematic representation of IgM and IgD interactions with CD19 and CD38 upon BCR activation, detailing the distinct inhibitory impacts of DARA and ISA.

Article Snippet: The CD38 cyclase assay was conducted using the CD38 Inhibitor Screening Assay Kit (BPS Bioscience, San Diego, California, USA) according to the manufacturer's protocol.

Techniques: Microscopy, Activation Assay

Journal: Clinical & Translational Immunology

Article Title: Targeting CD38 with monoclonal antibodies disrupts key survival pathways in paediatric Burkitt's lymphoma malignant B cells

doi: 10.1002/cti2.70011

Figure Lengend Snippet: Anti‐CD38 mAbs impair PI3K pathway signalling in pBL cells. (a, b) Time‐course analysis of SYK phosphorylation levels post anti‐CD38 mAb treatment over 24 h. (c) Comparative kinetic analysis of SYK dephosphorylation between DARA and ISA‐treated cells. (d, e) The phosphorylation status of AKT over a 24‐h period post anti‐CD38 mAb treatment. (f) Differential analysis of pAKT level kinetics between DARA and ISA treatments. Statistical significance was assessed using one‐way ANOVA with the post hoc Tukey test comparing untreated vs treated, indicated by: * P < 0.05; ** P < 0.01; *** P < 0.001. In (c) and (f) , the statistical significance was assessed using the unpaired t ‐test for the comparison between DARA vs ISA in each time point and the paired t ‐test for the comparison of the whole kinetic, and indicated by * P < 0.05; *** P < 0.001. Data are presented as mean ± SD. Non‐significance is not denoted. Data are representative of two independent experiments. All results were normalised and merged for consistent interpretation. (g) Schematic representation of the proposed effect of anti‐CD38 mAb (ISA) treatment on MYC/PI3K pathways and consequences on the proliferation/apoptosis balance.

Article Snippet: The CD38 cyclase assay was conducted using the CD38 Inhibitor Screening Assay Kit (BPS Bioscience, San Diego, California, USA) according to the manufacturer's protocol.

Techniques: De-Phosphorylation Assay, Comparison