Journal: iScience

Article Title: cGAS/STING sensing in dendritic cells discriminates between daptomycin sensitive and resistant Staphylococcus aureus clinical isolates

doi: 10.1016/j.isci.2026.115854

Figure Lengend Snippet: TLR2 is required for MutuDC sensing of, but not internalization of MRSA (A) Relative pHrodo labeled MRSA internalization by MutuDC over 4 h following stimulation with DapS A8819 (light blue symbols) or DapR A8817 (dark blue symbols), or media alone (white squares). Prior to stimulation, MutuDC were pre-treated for 1 h with TLR2 blocking antibody (clone T2.5; triangles with dashed lines) or media alone (circles with filled lines). Relative MRSA internalization by each DC subset is expressed as the gMFI of pHrodo. Results show the mean and (SD) of duplicates from one experiment, representative of two independent experiments. (B) Cytokine secretion (pg/mL) by MutuDC stimulated with TLR2 ligand peptidoglycan of S. aureus (PGN-SA) (10 μg/mL) or (C) DapS (A8819; light blue) or DapR (A8817; dark blue) MRSA (MOI of 10) for 18 h. MutuDC were first pre-treated with either TLR2 blocking antibody (dot-filled bars) or media alone (filled bars) as in A, or an isotype control (clone 163D3, empty bars) at 1 μg/mL. Results pooled from four (B) or three (C) independent experiments and expressed as the mean ± SEM, with each symbol (circle, square, and directional triangles) representing paired experimental replicates ( n = 3). Statistical significance determined using paired t test and reported as indicated by an ∗ when p ≤ 0.05. (D) Expression of surface activation markers by MutuDC stimulated with DapS A8819 MRSA. DC were pre-treated with TLR2 blocking antibody (black trace), isotype control (dashed red trace), and media alone (light blue shaded). Unstained control sample is shown for each marker (black dashed trace). Data shown from one experiment, representative of three independent experiments.

Article Snippet: mAB mTLR2- anti-mouse/human TLR2 , InvivoGen , Cat# mab-mtlr2; RRID: AB_763722.

Techniques: Labeling, Blocking Assay, Control, Expressing, Activation Assay, Marker

Journal: Bioactive Materials

Article Title: Injection site dictates the immune response to a biodegradable polymer and corresponding collagen regeneration

doi: 10.1016/j.bioactmat.2026.04.004

Figure Lengend Snippet: ID tissue exhibits higher densities of fibroblasts, macrophages, and microvessels compared to SC tissue. Immunofluorescence analysis comparing the expression of Vimentin (a fibroblast marker), CD68 (a macrophage marker), and CD31 (an endothelial cell marker) in ID and SC tissues. For each group, n = 3; data represent mean ± s.d.; ∗P < 0.05 and ∗∗P < 0.01.

Article Snippet: After antigen retrieval and blocking, sections were incubated overnight at 4 °C with primary antibodies targeting vimentin, CD68 (ABclonal, A20803), CD31 (R&D SYSTEMS, AF3628), HMGB1 (Cell Signaling Technology, 3935), HSP70 (ABclonal, A23457), NF-κB p65 (ABclonal, A19653), CD206 (Cell Signaling Technology, 24595), and FAPα (ABclonal, A23789 ).

Techniques: Immunofluorescence, Expressing, Marker

Journal: Bioactive Materials

Article Title: Glucose/ROS-responsive and redox-gated adaptive hydrogel dressing for accelerating diabetic wound repair via synergistic cGAS/STING pathway inhibition and oxidative stress alleviation

doi: 10.1016/j.bioactmat.2026.03.025

Figure Lengend Snippet: The cellular uptake and anti-inflammatory effect of HPSL in vitro . (A) Flow cytometry analysis and (B) semi-quantitative analysis of cellular uptake of PSL and blank NPs by M1 macrophages. n = 3. (C) Representative Giemsa staining images of LPS and high glucose-stimulated RAW 264.7 cells with different formulations, scale bar = 50 μm. (D) Immunofluorescence staining and semi-quantitative analysis of CD68 (red) and iNOS (green) in RAW 264.7 cells from different treatment groups, scale bar = 50 μm. n = 6. (E) Immunofluorescence staining and semi-quantitative analysis of CD68 (green) and Arg-1 (red) in RAW 264.7 cells from different treatment groups, scale bar = 50 μm. n = 6. Western blotting analysis and corresponding semi-quantitative analysis of (F) STING/ p -STING, (G) TBK1/ p -TBK1, (H) IRF3/ p -IRF3, (I) NF-κB, (J) TNF-α, and (K) IL-6, Lane 1: Normal group, Lane 2: Model group, Lane 3: PSL group, Lane 4: Free H151 group, Lane 5: HPSL group. n = 3. All data are shown as mean ± SEM.

Article Snippet: CD68-specific antibodies were purchased from Proteintech Group, Inc. (Wuhan, China).

Techniques: In Vitro, Flow Cytometry, Staining, Immunofluorescence, Western Blot

Journal: Bioactive Materials

Article Title: Translational selenium nanoparticles trigger apoptosis in triple-negative breast cancer cells through the MAPKs/Bcl2 pathway

doi: 10.1016/j.bioactmat.2026.02.027

Figure Lengend Snippet: Schematic illustration of PTR-SeNPs and MUC1@PTR-SeNPs synthesis and their anti-tumor efficacy against human triple-negative breast cancer.

Article Snippet: Anti-human MUC1 therapeutic antibody Fab fragment (7B8) were purchased From Creative Biolabs (TAB-423MZ-F, USA).

Techniques:

Journal: Bioactive Materials

Article Title: Translational selenium nanoparticles trigger apoptosis in triple-negative breast cancer cells through the MAPKs/Bcl2 pathway

doi: 10.1016/j.bioactmat.2026.02.027

Figure Lengend Snippet: Structure characterization of PTR-SeNPs and MUC1@ PTR-SeNPs. Structure characterization of PTR-SeNPs by (A) TEM, (B) Zetasizer Nano ZS, (C, D) Nanosight NS300, (E1-4) HRTEM-EDS and (F, G) FT-IR. (H) Confirmation of MUC1-C + PTR-SeNPs conjugation by confocal microscopy after fluorescent labeling with anti-mouse IgG (H + L). (I, J) Characterization results of the particle size and potential of MUC1@PTR-SeNPs

Article Snippet: Anti-human MUC1 therapeutic antibody Fab fragment (7B8) were purchased From Creative Biolabs (TAB-423MZ-F, USA).

Techniques: Conjugation Assay, Confocal Microscopy, Labeling

Journal: Bioactive Materials

Article Title: Translational selenium nanoparticles trigger apoptosis in triple-negative breast cancer cells through the MAPKs/Bcl2 pathway

doi: 10.1016/j.bioactmat.2026.02.027

Figure Lengend Snippet: In vitro anti-tumor efficacy of PTR-SeNPs and MUC1@PTR-SeNPs on 17 TNBC c ell lines. ( A, B ) Protein expression level of MUC1 in 17 different TNBC cell lines. ( C, D ) IC 50 and maximum % growth inhibition of PTR-SeNPs and MUC1@PTR-SeNPs on 17 TNBC cell lines. ( E, G ) Cell cycle distribution triggered by PTR-SeNPs and MUC1@PTR-SeNPs in HCC1937 and MDA-MB-436 cells. After treatment with PTR-SeNPs or MUC1@PTR-SeNPs (4 and 40 μM) in HCC1937 and MDA-MB-436 cells for 72 h, cells were stained with propidium iodide followed by flow cytometry analysis using MultiCycle software. The apoptotic cell death was quantified by measuring the sub-G1 cell population. ( F, H ) Phosphatidylserine translocation mediated by PTR-SeNPs and MUC1@PTR-SeNPs in HCC1937 and MDA-MB-436 cells. After treatment with MUC1@PTR-SeNPs (4 and 40 μM) for 48 h, cells were co-stained with propidium iodide and Annexin-V-FITC followed by flow cytometry analysis [early apoptotic subset: Annexin V+/PI- (green); late apoptotic subset: Annexin V+/PT+ (red)].

Article Snippet: Anti-human MUC1 therapeutic antibody Fab fragment (7B8) were purchased From Creative Biolabs (TAB-423MZ-F, USA).

Techniques: In Vitro, Expressing, Inhibition, Staining, Flow Cytometry, Software, Translocation Assay

Journal: Bioactive Materials

Article Title: Translational selenium nanoparticles trigger apoptosis in triple-negative breast cancer cells through the MAPKs/Bcl2 pathway

doi: 10.1016/j.bioactmat.2026.02.027

Figure Lengend Snippet: In vivo anti-tumor efficacy of MUC1@PTR- SeNPs. (A) MUC1 mRNA expression in normal tissue and primary breast cancer tumor using GEPIA database. ( B ) MUC1 expression in tumor tissues of MDA-MB-468-bearing mice in preliminary study. (C – E) Dose-dependent study of tumor inhibition effect of MUC1@PTR-SeNPs [75 (Low), 375 (Mid) & 750 μg (High) Se/kg BW/day] on BALB/c nude mice transplanted with MDA-MB-468 xenograft after oral administration for 30 days. PTR-SeNPs (High; 750 μg Se/kg BW/day) was used to investigate the possible improvement of in vivo anti-tumor efficacy by the MUC1@PTR-SeNPs. Quantitative analysis of Se content (μg/g) in (F) blood and (G) tumor tissue of experimental mice. (H) H&E, Ki67 and Tunnel fluorescence staining of tumor sections to detect apoptosis in vivo . (I) Western blot analysis of PARP, p-Bcl-2, Bax and C-caspase-9 protein expression in tumor sections. (J) In the serum of each group of tumor-bearing mice, the results of blood biochemistry-related indexes were analyzed.

Article Snippet: Anti-human MUC1 therapeutic antibody Fab fragment (7B8) were purchased From Creative Biolabs (TAB-423MZ-F, USA).

Techniques: In Vivo, Expressing, Inhibition, Fluorescence, Staining, Western Blot

Journal: Bioactive Materials

Article Title: Skin-mimetic bilayer hydrogel normalizes diabetic wound healing by orchestrating inflammatory cell dynamics: An early intervention strategy

doi: 10.1016/j.bioactmat.2026.02.025

Figure Lengend Snippet: In vitro assay of inflammation cell modulation under stimulation of SP-loaded Gel/HA and IL-10-loaded Ker/Cu. a Schematic of neutrophil migration test using a transwell system after treatment with the leaching solution of SP@Gel/HA. Gel/HA and blank culture medium were set for comparison. b Wright-Giemsa staining of HL-60 cells before and after differentiation. c Photograph of dHL-60 cells migrating to the lower chamber. d Quantitative analysis of neutrophil migration after treatments with SP@Gel/HA and Gel/HA. Untreated group serves as a control. e Schematic of macrophage polarization after treatment with LPS, IL-10, or IL-10/LPS. f Representative fluorescence images of macrophages after different treatment. Red: iNOS (M1 marker); Green: CD163 (M2c marker); Blue: DAPI (nuclear staining). g Schematic of macrophage efferocytosis test toward apoptotic dHL-60 cells under different treatments. h Flow cytometry plots of dHL-60 cells before and after apoptosis induction. i Representative fluorescent images of macrophage efferocytosis toward apoptotic dHL-60 cells under different treatments. Macrophages and apoptotic cells were stained green and red, respectively. All data were generated from at least three independent experiments and presented as the means ± standard deviation. Statistical analysis was performed by one-way ANOVA. ns, not significant; ∗∗∗∗p < 0.0001.

Article Snippet: After another 48 h, macrophage cells were harvested and stained with antibodies against human iNOS (Servicebio, GB11119) and CD163 (Abcam, ab182422) for 30 min. Fluorescence images were acquired using fluorescence microscope.

Techniques: In Vitro, Migration, Comparison, Staining, Control, Fluorescence, Marker, Flow Cytometry, Generated, Standard Deviation

Journal: Bioactive Materials

Article Title: Skin-mimetic bilayer hydrogel normalizes diabetic wound healing by orchestrating inflammatory cell dynamics: An early intervention strategy

doi: 10.1016/j.bioactmat.2026.02.025

Figure Lengend Snippet: Bilayer hydrogel orchestrates inflammatory cell dynamics during the early inflammation phase of diabetic wound healing. a Experimental timeline for assay of early neutrophil recruitment. b Immunohistochemical staining for Ly-6G in wounds at 8 h, 1 d and 3 d after injury. Diabetic wounds were treated with SP/IL-10@Bilayer, SP@Bilayer, IL-10@Bilayer, and saline solution (Model), respectively. Healthy mice treated with saline solution were set as Normal. c Quantitative analysis of Ly-6G + cells in each group. d Relative expression of CXCL-1 on day 1. e Relative expression of MCP-1 on day 1. f Experimental timeline for assay of M1 macrophage infiltration. g Immunofluorescence staining for iNOS in wounds on days 1, 3 and 6 after injury. h Quantitative analysis of iNOS + cells in each group. i-k Relative expressions of macrophage-associated pro-inflammatory cytokines including TNF-α, IL-1β and IL-6 on day 3. l Schematic illustrating the dynamic modulation of inflammatory cells during the early inflammation phase of diabetic wounds by SP/IL-10@Bilayer. All data were generated from at least three independent experiments and presented as the means ± standard deviation. Statistical analysis was performed by one-way ANOVA. # means significant difference compared to the normal group. #p < 0.05, ##p < 0.01 and ###p < 0.001; ∗ means significant difference compared to the model group. ∗p < 0.05; & means significant difference compared to SP/IL-10@Bilayer. & p < 0.05 and && p < 0.01.

Article Snippet: After another 48 h, macrophage cells were harvested and stained with antibodies against human iNOS (Servicebio, GB11119) and CD163 (Abcam, ab182422) for 30 min. Fluorescence images were acquired using fluorescence microscope.

Techniques: Immunohistochemical staining, Staining, Saline, Expressing, Immunofluorescence, Generated, Standard Deviation

Journal: iScience

Article Title: Integrating complementary approaches reveals antigen-reactive CD4 + T cell states after SARS-CoV-2 vaccination

doi: 10.1016/j.isci.2026.116175

Figure Lengend Snippet: Identification of SARS-CoV-2-specific T cell responses by reverse phenotyping (A) CoVa-Adapt study design and sample collection scheme. For all donors, PBMCs were collected at day 0 (P0), 10 days after primary (P10), 10 and 210 days after secondary (S10, S210), and 10 and 189 days after tertiary (T10, T189) vaccination. For selected donors, PBMCs were additionally sampled 108 days after tertiary vaccination (T108, n = 7). Vaccination-induced T cell responses were characterized for most donors on a quantitative level by IFNγ ELISpot. Selected CoVa-Adapt donors were subjected to in-depth characterization using scRNAseq (reverse phenotyping and epitope-specific analyses) followed by TCR functional testing. (B–G) scRNAseq data from the reverse phenotyping dataset. For reverse phenotyping, PBMCs were re-stimulated with 15-mer peptides covering the complete wild-type spike protein or left untreated. Sorted non-naïve CD4 + and/or CD8 + T cells were subjected to scRNAseq. Only CD4 + T cells are shown (annotation described in the methods section). The full dataset is depicted in . (B) UMAP of stimulated (blue) and unstimulated (orange) T cells (left) and Leiden clusters (right; cluster names in UMAP, cluster numbers on the right) ( n = 101,939 cells in total). Cells located within the reactive cluster are displayed with increased point size. (C) Dot plots of log-normalized expression of representative genes per cluster. Selected genes of the reactive cluster are highlighted in gray. Numbers on the left indicate cluster numbers with reactive cluster 12 highlighted in bold. (D and E) IFNG expression (D) and proliferation score (E) in unstimulated (stimulated cells in gray) and stimulated (unstimulated cells in gray) CD4 + T cells (left), and quantification in the stimulated condition of cells in the reactive cluster versus all other clusters (right). Cells located within the reactive cluster are displayed with increased point size. For IFNG , cells with log-normalized gene expression of 0 are shown in gray in UMAPs. Statistical testing by the Mann-Whitney U test. (F) UMAP visualization of cells classified as reactive (cells located in the reactive cluster or belonging to clones where at least one cell is in the reactive cluster) from donor A5 at individual time points after primary (P), secondary (S), and tertiary (T) vaccination in the stimulated condition. Color gradient indicates IFNG expression at the indicated time points. Non-reactive cells and cells from other donors are shown in gray. (G) Fraction of cells from donor A5 at each time point belonging to the reactive cluster. (H and I) Identification of spike-reactive T cells after 20h of in vitro re-stimulation of PBMCs with 15-mer peptides covering the complete wild-type spike protein. Peptides were provided in two subpools, S1 (depicted in H) and S2. Primary data (H) of donor A5 is shown. Quantification (I) of spot-forming units (SFU) for IFNγ ELISpot (combined frequencies of S1 and S2 subpools), data points represent individual donors ( n = 12–19 per time point), solid lines indicate the mean. Samples without SFU above the negative control were set to not detected (n.d.). Donor A5 is highlighted in pink. Statistical testing by the Kruskal-Wallis test followed by Dunn’s multiple comparisons test. Significant differences from the P10 time-point are indicated. ∗∗ p < 0.01, ∗∗∗∗ p < 0.0001, n.s. not significant.

Article Snippet: Plates were washed with PBS containing 0.05% Tween 20 (Sigma-Aldrich, P9416-50 mL) and incubated with biotinylated anti-human IFNγ monoclonal antibody (clone 7-B6-1, Mabtech, 3420-6-250) at 0.2 μg/well for 2 h. Plates were washed a second time with PBS containing 0.05% Tween 20 and subsequently incubated with an avidin-biotinylated peroxidase complex (VECTASTAIN Elite ABC-HRP Kit, Vector Laboratories, VEC-PK-6100) for 1–2 h. Afterward, plates were washed first with PBS containing 0.05% Tween 20 following one washing step with PBS.

Techniques: Enzyme-linked Immunospot, Functional Assay, Expressing, Gene Expression, MANN-WHITNEY, Clone Assay, In Vitro, Negative Control