Journal: Frontiers in Immunology

Article Title: Unveiling spatial complexity in solid tumor immune microenvironments through multiplexed imaging

doi: 10.3389/fimmu.2024.1383932

Figure Lengend Snippet: Immunophenotyping panel for multiplexed tissue imaging of cancer.

Article Snippet: CD206 , DCN228 , 50 , 130-123-671 , FITC (PE) , Miltenyi Biotec.

Techniques: Imaging

Journal: Frontiers in Immunology

Article Title: Unveiling spatial complexity in solid tumor immune microenvironments through multiplexed imaging

doi: 10.3389/fimmu.2024.1383932

Figure Lengend Snippet: Deep spatial profiling of human palatine tonsil tissues. (A) Hematoxylin and eosin (H&E) staining after 92 MICS cycles, including the marked epithelium, germinal center (GC), and T cell zone of the lymphoid follicle. MICS DAPI and stroma staining depicting the composition and structure of the tonsil. Markers: Collagen III, collagen IV, fibronectin (all extracellular matrix (ECM), cytokeratin (epithelium), podoplanin (lymphatic vessels), CD105/SM Actin (blood vessels). (B) Immune cell content of a human palatine tonsil comprising T cells (CD3), B cells (CD19/CD20), plasma cells (PCs) (CD38/CD138), NK cells (CD56), granulocytes (CD15/CD66b), mast cells (CD117), macrophages (MΦ) (CD163/CD169/CD206), myeloid dendritic cells (mDCs) (CD11c), and plasmacytoid dendritic cells (pDCs) (CD123). (C) Detailed view on the T cell zone, mainly composed of CD4 + helper T cells (T h ) and CD8 + cytotoxic T cells (T c ), mDCs (CD11c), and PCs (CD38/CD138). (D) Detailed view on the GC-mantle zone border, showing different B cells (CD11b, CD21, CD22), mDCs (CD11c), and PCs (CD38/CD138). (E) Cell annotations of three different tonsil samples plus respective bar graphs of gated cell populations, comparing the cell content between the three tonsil samples. Depicted markers and annotated cell types as indicated by the color code. ROI sizes: 976 x 640 µm, zoomed-in subregions in (C, D) : 334 µm x 219 µm. Scale bar: 100 µm.

Article Snippet: CD206 , DCN228 , 50 , 130-123-671 , FITC (PE) , Miltenyi Biotec.

Techniques: Staining, Clinical Proteomics

Journal: Frontiers in Immunology

Article Title: Arginine-Based Poly(I:C)-Loaded Nanocomplexes for the Polarization of Macrophages Toward M1-Antitumoral Effectors

doi: 10.3389/fimmu.2020.01412

Figure Lengend Snippet: Polarization of M0 and M2 macrophages after treatment with free and nanocomplexed poly(I:C). Expression of the M2 markers (A–D) CD206 and (E–H) CD163 in ENCP-treated M0 and M2 macrophages, in comparison to the prototypic phenotypes evaluated by FACS. M2 M1 represents M2 macrophages that were treated with LPS + IFN-γ for their M1 polarization. Macrophages were incubated with the treatments for 48 h, and the poly(I:C) dose used was 5 μg/mL. Each symbol shape represents a different donor. Values are shown as mean ± SD ( n ≥ 3). Statistical comparison was done using an ordinary one-way ANOVA followed by a Tukey's comparison test between groups; or a paired t -test for (C,G) . Statistically significant differences are represented as ** p < 0.01. C12r8, laurate-octaarginine; HA, hyaluronic acid; pArg, poly-arginine; PEG–PGA, pegylated polyglutamic acid; pIC, poly(I:C).

Article Snippet: They were then stained with APC-mouse anti-human HLA-DR (552764, BD Biosciences, CA, USA), FITC-mouse anti-human mannose receptor CD206 (551135), anti-human CD163-BV421 (562643), CD80 APC-H7-mouse anti-Human CD80 (Clone L307.4; 561134) and anti-human CD68-PE (556078) (all from BD Biosciences, CA, USA).

Techniques: Expressing, Incubation

Journal: Frontiers in Immunology

Article Title: Arginine-Based Poly(I:C)-Loaded Nanocomplexes for the Polarization of Macrophages Toward M1-Antitumoral Effectors

doi: 10.3389/fimmu.2020.01412

Figure Lengend Snippet: Schematic illustration of the in vitro effects of the poly(I:C) ENCPs developed in this study. (A) Upon interaction with macrophages, poly(I:C) nanocomplexes are taken up. (B) This process allows poly(I:C) to reach its target receptor TLR3, found in the endosomes. It is expected that this interaction activates the TLR3 and the TRIF pathway, stimulating the upregulation of type I IFN genes. (C) The expression of M2 (CD206 and CD163) surface markers was slightly decreased, while M1 (CD80 and MHC II) markers are not substantially modified. Nevertheless, (D) CXCL10 and CCL5 chemokines, involved in the attraction of CD8 T cells to the tumor microenvironment, are secreted. (E) The direct cytotoxicity of macrophages toward cancer cells is also enhanced. Images were reproduced from Servier Medical Art under a Creative Commons Attribution 3.0 Unported License https://creativecommons.org/licenses/by/3.0 .

Article Snippet: They were then stained with APC-mouse anti-human HLA-DR (552764, BD Biosciences, CA, USA), FITC-mouse anti-human mannose receptor CD206 (551135), anti-human CD163-BV421 (562643), CD80 APC-H7-mouse anti-Human CD80 (Clone L307.4; 561134) and anti-human CD68-PE (556078) (all from BD Biosciences, CA, USA).

Techniques: In Vitro, Expressing, Modification

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: Advanced science (Weinheim, Baden-Wurttemberg, Germany)

Article Title: Mucous Permeable Nanoparticle for Inducing Cuproptosis-Like Death In Broad-Spectrum Bacteria for Nebulized Treatment of Acute Pneumonia.

doi: 10.1002/advs.202408580

Figure Lengend Snippet: Figure 8. Activation of macrophage response by Cu2O-BSO NPs in vitro. a) Typical scatter plots of MH-S surface markers CD86 (M1 macrophage marker) and CD206 (M2 macrophage marker) detected by flow cytometry. b) Typical images of MH-S engulfing a bacterium. The green fluorescence represents GFP-MRSA, the red fluorescence represents MH-S, and the blue fluorescence represents the nucleus. The stronger the green fluorescence, the stronger the phagocytosis of MH-S. c,d) Representative SPM photographs and CFU count of MRSA engulfed by MH-S. e–g) ELISA results indicating levels of cytokines (IL-1𝛽, IL-6, and TNF-𝛼) secreted by MH-S in different groups.

Article Snippet: Each group of cells was centrifuged, washed, and resuspended in 500 μL of PBS, which contained an anti-mouse CD86 antibody coupled with FITC (Elabscience) and an anti-mouse CD206 antibody coupled with PE (Elabscience).

Techniques: Activation Assay, In Vitro, Marker, Cytometry, Enzyme-linked Immunosorbent Assay