Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Association of CCL21 + CAFs with B-cell recruitment and TLS maturation in penile squamous cell carcinoma

doi: 10.1007/s00262-026-04348-5

Figure Lengend Snippet: Evaluation of TLS maturation and prognostic analysis of PSCC patients. A UMAP plot of the cell types identified in PSCC through spatial transcriptomic sequencing, depicting 7 distinct cell populations. B Dot plot displaying the expression of cell type-specific markers. The dot size represents the proportion of cells expressing the marker, while the colour intensity indicates the mean expression level. C Regions with AUCell scores for mTLS genes ( BCL6 , AICDA , CD38 , ICOS , CXCR5 , CXCL13 , and CD21 ) above the median were defined as mTLSs, whereas those below the median were classified as iTLSs. D Spatial visualization of AUCell scores for mTLS genes, where warmer colours indicate higher scores. E Spatial distribution of iTLSs (green) and mTLSs (orange) across different regions of PSCC tissues. F Representative images H&E staining and immunofluorescence staining for CD20 (green) and CD21 (red) in iTLSs and mTLSs. G GO enrichment analysis of upregulated genes in mTLS. Darker colours denote more significant enrichment of the pathway, and larger dots represent a greater number of genes involved. H Kaplan–Meier survival curves showing that patients with mTLSs experienced significantly longer overall survival than those with iTLSs, as determined using ssGSEA ( P < 0.05)

Article Snippet: The following antibodies were used: CD20 (Proteintech, 60,271–1-Ig, 1:500), CD21 (Proteintech, 28,206–1-AP, 1:250), ACTA2/α-SMA (Sigma, SAB5700835, 1:300), and CCL21 (Abcam, ab9851, 1:300).

Techniques: Sequencing, Expressing, Marker, Staining, Immunofluorescence

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Association of CCL21 + CAFs with B-cell recruitment and TLS maturation in penile squamous cell carcinoma

doi: 10.1007/s00262-026-04348-5

Figure Lengend Snippet: Characterization of CCL21 + CAF subtypes and their association with TLS maturation in PSCC. A UMAP plot of CAF subtypes in PSCC derived from spatial transcriptomics, including CCL21 + CAFs (green), COL16A1 + CAFs (blue), MYH11 + CAFs (orange), and XBP1 + CAFs (red). B Dot plot of key markers identified across CAF subtypes. The dot size represents the proportion of cells expressing the marker, and colour intensity indicates the mean expression level. C Radar plot showing the distribution of CCL21 + CAF proportions across the iTLS-core, mTLS-core, iTLS-edge, and mTLS-edge regions. D Bar chart illustrating the spatial distances between different CAF subtypes and mTLS structures; ***, P < 0.001. E In situ visualization of TLS regions: green areas indicate the iTLS-core, blue circles indicate the iTLS-edge, orange areas represent the mTLS-core, and yellow circles denote the mTLS-edge. F Spatial localization of CCL21 + CAFs in PSCC tissues showing their association with distinct TLS regions. G Heatmap overlay of the CCL21 + CAF spatial density and TLS distribution, with red indicating a higher CCL21 + CAF abundance. H Scatter plots of the correlation between the CCL21 + CAF proportion and B-cell (left panel) and naive T-cell (right panel) numbers in the iTLS/mTLS regions. I Representative images of mIHC of TLS regions in PSCC showing the spatial distribution and colocalization of CD20 (purple), ACTA2/α-SMA (green), and CCL21 (yellow). Scale bar = 150 μm. ( J ) Patients with high CCL21 + CAF signature scores (n = 29) experienced significantly longer overall survival than those with low scores (n = 26)

Article Snippet: The following antibodies were used: CD20 (Proteintech, 60,271–1-Ig, 1:500), CD21 (Proteintech, 28,206–1-AP, 1:250), ACTA2/α-SMA (Sigma, SAB5700835, 1:300), and CCL21 (Abcam, ab9851, 1:300).

Techniques: Derivative Assay, Spatial Transcriptomics, Expressing, Marker, In Situ

Journal: Materials Today Bio

Article Title: Sophora flavescens-derived extracellular vesicles loaded with oncolytic vaccinia virus/IR1061 for NIR-II photoacoustic imaging guided multimodal treatment of diffuse large B-cell lymphoma

doi: 10.1016/j.mtbio.2025.102177

Figure Lengend Snippet: SFOVV@IR1061 induces apoptosis and ICD in A20 lymphoma cells. (A) Flow cytometric analysis of apoptosis in A20 cells treated with PBS, SFNPs, OVV, IR1061 + laser, OVV@IR1061+laser, or SFOVV@IR1061 + laser (1064 nm, 5 min, 0.8 W/cm 2 ) for 12 h. Cells were stained with Annexin V-FITC and propidium iodide (PI). (B) Representative CLSM images of live/dead cell staining using Calcein-AM (green, live cells) and PI (red, dead cells) under the same treatment conditions. (C) Quantification of the dead/live cell ratio from CLSM images shown in (B). Data are presented as mean ± SD (n = 3). (D) Western blot analysis of apoptosis-related proteins including PARP, and Caspase-3 in A20 cells treated as indicated. β-actin was used as a loading control: 1. PBS; 2. SFNPs; 3. OVV; 4. IR1061+laser; 5. OVV@IR1061+laser; 6. SFOVV@IR1061+laser. (E) Western blot analysis of ICD-related markers CRT, HSP70, and HMGB1 following the same treatments: 1. PBS; 2. SFNPs; 3. OVV; 4. IR1061+laser; 5.OVV@IR1061 +laser; 6. SFOVV@IR1061+laser. (F) Flow cytometric analysis of macrophage polarization markers CD86 (left) and CD206 (right) in RAW264.7 cells. (G) Quantification of fluorescence intensity for CD86 and CD206. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: To analyze M1/M2 macrophage polarization, cells were incubated with FITC Anti-Mouse CD86 Antibody (Elabscience, China) to detect M1 polarization and APC Anti-Mouse CD206 Antibody (Elabscience, China) to detect M2 polarization, according to the manufacturer's instructions.

Techniques: Staining, Western Blot, Control, Fluorescence

Journal: Cold Spring Harbor Molecular Case Studies

Article Title: Tumor-immune microenvironment revealed by Imaging Mass Cytometry in a metastatic sarcomatoid urothelial carcinoma with a prolonged response to pembrolizumab

doi: 10.1101/mcs.a006151

Figure Lengend Snippet: Full list of antibodies and their properties

Article Snippet: CD20 , Dy161 , L26 , 1:200 , 0.5 , Novus Biologicals , NBP2-80486.

Techniques: Concentration Assay

Journal: bioRxiv

Article Title: BCG vaccination elicits protection against Mtb infection mediated by two phases of T cell immunity

doi: 10.1101/2025.10.28.685186

Figure Lengend Snippet: A. Experimental scheme. CC042 mice were treated with anti-CD20 depletion mAbs one day before and five weeks after BCG vaccination. Mice were rested for 12 weeks after vaccination, and then all mice were infected with Rv.YFP for four weeks. B. Representative flow plots (left) showing CD19 (B cells) and CD3 (T cells) expression of total live cells in the blood of BCG vaccinated versus BCG vaccinated and B cell depleted CC042 mice and frequency (right) of CD19+ B cells in blood four weeks after BCG vaccination of CC042, as determined by flow cytometry. Data is from one of two independent experiments with similar results. One-way ANOVA with Šídák’s multiple comparisons test. Each point represents an individual subject, n=5 mice per group. Bars, mean. Error bars, SD. C. Quantification of total serum antibodies specific to CFP (left) and WCL (right) at the endpoint. One-way ANOVA with Šídák’s multiple comparisons test. Each point represents the mean of technical duplicate replicates, n=5 mice per group. Box plots indicate median (middle line), 25th, 75th percentile (box) and minimum and maximum (whiskers). Data is from one of two independent experiments with similar results. D. CFU in the lung (left) and spleen (right) at four wpi. One-way ANOVA with Šídák’s multiple comparisons test. Each point represents an individual subject, n=4-5 mice per group. Bars, mean. Error bars, SD. Data is from one of two independent experiments with similar results.

Article Snippet: CD20: Mice were given 250 μg of anti-CD20 mAb (MB20-11, Bio X Cell) suspended in 200 μL of PBS via the intraperitoneal route one week before vaccination and again six weeks later (five weeks post-vaccination).

Techniques: Infection, Expressing, Flow Cytometry

Journal: medRxiv

Article Title: Spatial mapping of Ethiopian cutaneous leishmaniasis lesions reveals distinct tissue level immune programs

doi: 10.64898/2026.02.04.26345554

Figure Lengend Snippet: A , Heatmap of top 100 DEGs in P1_L vs other lesional skin (P2-P5 L) (padj < 0.05, log2FC > ±0.25; Wilcoxon) B , UMAP of cytotoxicity signature in lesional skin (P1-P5). Top: composite score; right/below: four cytotoxic effector genes. C , Cytotoxicity score across all patients (Wilcoxon; P1_L vs P2-P5 L). D , Spatial map of cytotoxicity score in P1_L (middle), underlying H&E histology (left panel) and zoomed view of a representative area showing individual gene expression for selected genes (right). E, Cell type deconvolution (pie charts) of representative area (from D ). F , Fold enrichment of topics in high (≥Q3, top 25%) vs low (

Article Snippet: For separate experiment, sections were stained with CD20 (IGEL/773, NBP2-47840C, Novus Biologicals, 1:100, Dylight-650) for B cells and CD3E (CD3-12, ab11089, Abcam, 1:300]) for T cells.

Techniques: Gene Expression, Multiplex Assay, Immunofluorescence