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: CD3 , REA1151 , 50 , 130-120-267 , FITC (APC) , Miltenyi Biotec.

Techniques: Imaging

Journal: Journal of Translational Medicine

Article Title: Niraparib exhibits a synergistic anti-tumor effect with PD-L1 blockade by inducing an immune response in ovarian cancer

doi: 10.1186/s12967-021-03073-0

Figure Lengend Snippet: The information of antibodies used for western blot, immunohistochemistry, and flow cytometry

Article Snippet: CD3 and isotype , Human , 5 μl of antibody per test , Elabscience, E-AB-F1001E , Flow cytometry.

Techniques: Western Blot, Immunohistochemistry, Flow Cytometry

Journal: The Journal of clinical endocrinology and metabolism

Article Title: Association of postpartum thyroid dysfunction with antepartum hormonal and immunological changes.

doi: 10.1210/jc.2002-021219

Figure Lengend Snippet: FIG. 1. Graphical output from the FACScan analyzer (Becton Dickinson and Co.). Before analysis lymphocytes were stained with R-phyco- erythrin-Cy5 mouse antihuman CD3 fluorescein isothiocyanate-conjugated mouse antihuman CD4 together with either phycoerythrin- conjugated rat antihuman IL-4 or phycoerythrin-conjugated mouse antihuman IFN-. The output was gated on the basis of side scatter and CD3 staining. Shown are control cells in the left panels and stimulated cells in the right panels. Lymphocytes gated for right-angle light scatter and CD3 staining (a and b), CD4 expression (c and d), IL-4 expression (c–f), and IFN- expression (e and f) are shown.

Article Snippet: The culture plate was placed in a humidified incubator at 37 C, gas phase 5% CO2 in air, for 4 h. Aliquots (350 l) from the stimulated and unstimulated wells were transferred into fluorescenceactivated cell sorter (FACS) tubes and the surface antigen (CD3 and CD4) was stained by adding 5 l conjugated antibody (R-phycoerythrin-Cy5 conjugated mouse antihuman CD3, DAKO Corp. Ltd., Ely, UK; fluorescein isothiocyanate-conjugated mouse antihuman CD4, Serotec Ltd., Oxford, UK) and incubated in the dark for 30 min.

Techniques: Staining, Control, Expressing

Journal: Molecular Cancer Research

Article Title: Neoadjuvant Chemotherapy Induces IL34 Signaling and Promotes Chemoresistance via Tumor-Associated Macrophage Polarization in Esophageal Squamous Cell Carcinoma

doi: 10.1158/1541-7786.mcr-20-0917

Figure Lengend Snippet: Figure 1. Increased infiltration and polarized differentiation of TAMs by NAC in ESCC. A, Schematic illustration of the experimental design. B, Representative IHC images of CD163 expression in ESCC specimens from patients who received NAC [NAC (þ)] and did not receive NAC [NAC ()]. Scale bars, 100 mm; original magnification 200. Pre-, Pre-NAC; Post-, Post-NAC. C, Comparison of the number of CD163þ TAMs between biopsy and surgically resected ESCC specimens in NAC () (n ¼ 12) and NAC (þ) (n ¼ 33). D, Comparison of the number of CD163þ TAMs between NAC () (n ¼ 12) and NAC (þ) (n ¼ 33) in biopsy and surgically resected ESCC specimens. E, Comparison of the intensities of CD163 expression between biopsy and surgically resected ESCC specimens in NAC () (n ¼ 12) and NAC (þ) (n ¼ 33). F, Comparison of the intensities of CD163 expression between NAC () (n ¼ 12) and NAC (þ) (n ¼ 33) in biopsy and surgically resected ESCC specimens. G, Representative IHC images of the expression of CD163, CD3, CD8, CD105, E-cadherin, and vimentin in surgically resected ESCC specimens from patients who received NAC [NAC (þ)]. Scale bars, 100 mm; original magnification 200. Post-, Post-NAC. H, Correlations between the number of CD163þ cells and the number of CD3þ TILs, the number of CD8þ TILs, CD105þ microvessel counts, E-cadherin staining score, or Vimentin staining score in surgically resected ESCC specimens of NAC (þ) (n ¼ 33). , P < 0.01; , P < 0.001; , P < 0.0001.

Article Snippet: Reagents used in this study were acquired from the indicated suppliers: 5-FU, KU-55933, and VE-821 (Merck Sigma-Aldrich); cisplatin (FUJIFILM Wako Pure Chemical Corporation); antihuman IL34 antibody [1D12] (ab101443), anti-M-CSF antibody [EP1179Y] (ab52864; Abcam); Novocastra Liquid Mouse Monoclonal Antibody CD163 (NCL-L-CD163; Leica Biosystems); anti-human CD3 antibody (IR503), anti-human CD8 (M7103), and E-cadherin mAb (NCH-38; Dako/Agilent Technology); anti-human CD105 antibody (28117-1-AP; Proteintech); vimentin mAb (SP20; Nichirei Bioscience); antibody APC/Cyanine7 anti-human CD14 [HCD14] (325619), PE anti-human CD163 [GHI/61] (333605; BioLegend); 7-AAD (559925; BD Biosciences); PLX-3397 [PLX; HY-16749], BLZ945 [BLZ; HY-12768] (MedChemExpress).

Techniques: Expressing, Comparison, Staining

Journal: Advanced Science

Article Title: Bispecific Nanosystems Enable Multieffector Immune Cell Retargeting for Hematologic Malignancy Therapy

doi: 10.1002/advs.202509103

Figure Lengend Snippet: A) This diagram depicts the dual‐targeting strategy of biHSNPs in facilitating tumor cell elimination. Each biHSNP features two functional arms: one specifically binds to a tumor‐associated antigen on lymphoma cells (CD19/CD20), while the other engages and activates immune effector cells (CD3/CD16). By bridging tumor cells and immune cells, this approach enhances immune‐mediated tumor cell lysis, leveraging the immune system's inherent ability to identify and eliminate abnormal cells for precise and efficient cancer cell killing. B) Schematic illustration of the T cell activation and tumor cell death mechanism mediated by biHSNPs.

Article Snippet: FITC anti‐human CD19 antibody[CB19] (catalog No.: E‐AB‐F1004C, species reactivity: human), FITC anti‐human CD20 antibody[BCA/B20] (Catalog No.: E‐AB‐F1045C, species reactivity: human) and FITC anti‐human CD3 antibody[OKT‐3] (catalog No.: E‐AB‐F1001C, species reactivity: human) were obtained from Elabscience Biotechnology Co.,Ltd. (China, Wuhan).

Techniques: Functional Assay, Lysis, Activation Assay

Journal: Advanced Science

Article Title: Bispecific Nanosystems Enable Multieffector Immune Cell Retargeting for Hematologic Malignancy Therapy

doi: 10.1002/advs.202509103

Figure Lengend Snippet: Schematic illustration of the chemical synthesis and characterization of antibody‐conjugated hollow silica nanoparticles. A) Synthetic pathway of PEGylated hollow silica nanoparticles. B) Fourier transform‐infrared spectroscopy (FT‐IR) characterization of HSNP@PEG (2). C) Proton nuclear magnetic resonance ( 1 H NMR) characterization of HSNP@PEG. D) DLS and TEM images of HSNP@PEG. E) DLS and TEM images of HSNP@PEG@NH 2 (3). F) Fluorescence spectrum of HSNP@PEG@NH 2 @FITC (9.62 µ m ), confirming successful FITC conjugation. G) DLS and TEM images of HSNP@PEG@NH 2 @FITC (4). H–K) DLS and TEM results for monospecific nanoparticles (HSNP αCD3 , HSNP αCD16 , HSNP αCD19 , HSNP αCD20 ). L–O) DLS and TEM results for bispecific nanoparticles (HSNP αCD3 + αCD19 , HSNP αCD3 + αCD20 , HSNP αCD16 + αCD19 , HSNP αCD16 + αCD20 ). All scale bars in the TEM images represent 100 nm. HSNP αCD3 , HSNP αCD16 , HSNP αCD19 and HSNP αCD20 : Monospecific HSNPs conjugated with a single type of antibody (anti‐CD3, anti‐CD16, anti‐CD19, or anti‐CD20, respectively). HSNP αCD3 + αCD19 , HSNP αCD3 + αCD20 , HSNP αCD16 + αCD19 and HSNP αCD16 + αCD20 : Bispecific HSNPs conjugated with two different types of antibodies (e.g., anti‐CD3 and anti‐CD19). Antibody types are indicated using lowercase Greek letters (e.g., αCD3).

Article Snippet: FITC anti‐human CD19 antibody[CB19] (catalog No.: E‐AB‐F1004C, species reactivity: human), FITC anti‐human CD20 antibody[BCA/B20] (Catalog No.: E‐AB‐F1045C, species reactivity: human) and FITC anti‐human CD3 antibody[OKT‐3] (catalog No.: E‐AB‐F1001C, species reactivity: human) were obtained from Elabscience Biotechnology Co.,Ltd. (China, Wuhan).

Techniques: Fourier Transform Infrared Spectroscopy, Spectroscopy, Nuclear Magnetic Resonance, Fluorescence, Conjugation Assay

Journal: Advanced Science

Article Title: Bispecific Nanosystems Enable Multieffector Immune Cell Retargeting for Hematologic Malignancy Therapy

doi: 10.1002/advs.202509103

Figure Lengend Snippet: Binding specificity and cell–cell linkage induced by biHSNPs. Target cells were incubated with FITC‐labeled, antibody‐coated HSNPs at 4 °C for 30 min, followed by flow cytometry analysis to confirm binding specificity. A–C) Depict cell–cell linkage induced by biHSNPs: A) Schematic illustration of cell‐cell complex formation mediated by biHSNPs. B) Confocal microscopy images showing stable junctional complexes between immune effector cells and tumor cells facilitated by HSNP αCD3 + αCD19 and HSNP αCD3 + αCD20 . C) Flow cytometry analysis of cell–cell complexes between pre‐stained Jurkat (green, CellTracker Green CMFDA) and Raji cells (red, CellTracker Red CMTPX), demonstrating effective linkage by biHSNPs. D) Verification of biHSNPs binding to CD3 + cells using PBMCs as the CD3 + cell line. E) Verification of biHSNPs binding to CD19 + /CD20 + cells using Raji cells as the target. F) Verification of biHSNPs binding to CD16 + cells using PBMCs as the CD16 + cell line. G) Verification of biHSNPs binding to CD19 + /CD20 + cells using Raji cells as the target.

Article Snippet: FITC anti‐human CD19 antibody[CB19] (catalog No.: E‐AB‐F1004C, species reactivity: human), FITC anti‐human CD20 antibody[BCA/B20] (Catalog No.: E‐AB‐F1045C, species reactivity: human) and FITC anti‐human CD3 antibody[OKT‐3] (catalog No.: E‐AB‐F1001C, species reactivity: human) were obtained from Elabscience Biotechnology Co.,Ltd. (China, Wuhan).

Techniques: Binding Assay, Incubation, Labeling, Flow Cytometry, Confocal Microscopy, Staining

Journal: Advanced Science

Article Title: Bispecific Nanosystems Enable Multieffector Immune Cell Retargeting for Hematologic Malignancy Therapy

doi: 10.1002/advs.202509103

Figure Lengend Snippet: A) Cytotoxicity and cytokine release assays of biHSNPs in luciferase‐transfected Raji cells. B) In vitro cytotoxicity of HSNP@PEG, HSNP αCD3 , HSNP αCD19 , and bispecific HSNP αCD3 + αCD19 . C) In vitro cytotoxicity of HSNP@PEG, HSNP αCD3 , HSNP αCD20 , and bispecific HSNP αCD3 + αCD20 . D,E) Cytokine secretion levels (IFN‐γ, IL‐2, and TNF‐α) for HSNP αCD3 , HSNP αCD19 , and HSNP αCD3 + αCD19 ; and HSNP αCD20 and HSNP αCD3 + αCD20 . F) In vitro cytotoxicity of HSNP@PEG, HSNP αCD16 , HSNP αCD19 , and bispecific HSNP αCD16 + αCD19 . G) In vitro cytotoxicity of HSNP@PEG, HSNP αCD16 , HSNP αCD20 , and bispecific HSNP αCD16 + αCD20 . H,I) Cytokine secretion levels (IFN‐γ, IL‐2, and TNF‐α) for HSNP αCD16 , HSNP αCD19 , HSNP αCD16 + αCD19 , HSNP αCD20 , and HSNP αCD16 + αCD20 . J) Cytotoxicity of combined bispecific nanoparticle treatments: HSNP αCD3 + αCD19 with HSNP αCD16 + αCD19 , and HSNP αCD3 + αCD20 with HSNP αCD16 + αCD20 . K) In vitro cytotoxicity of HSNP αCD3 + αCD19 combined with HSNP αCD16 + αCD19 , and HSNP αCD3 + αCD20 combined with HSNP αCD16 + αCD20 . Data are presented as mean ± SD ( n ≥ 3), Statistical differences in B–I) were analyzed by One‐way ANOVA and K) were analyzed by Student's t‐test and the statistical significance is indicated as ** P ≤ 0.01 and *** P ≤ 0.001.

Article Snippet: FITC anti‐human CD19 antibody[CB19] (catalog No.: E‐AB‐F1004C, species reactivity: human), FITC anti‐human CD20 antibody[BCA/B20] (Catalog No.: E‐AB‐F1045C, species reactivity: human) and FITC anti‐human CD3 antibody[OKT‐3] (catalog No.: E‐AB‐F1001C, species reactivity: human) were obtained from Elabscience Biotechnology Co.,Ltd. (China, Wuhan).

Techniques: Luciferase, Transfection, In Vitro