antihuman cd20 fitc  (Miltenyi Biotec)

Journal: Advanced Science

Article Title: Spatial Transcriptional Dynamics of CD74⁺ B Cells in Tertiary Lymphoid Structures Drive Immune Evolution in Penile Squamous Cell Carcinoma

doi: 10.1002/advs.202509742

Figure Lengend Snippet: Gene expression in and functional analysis of CD74⁺ B cells within and outside TLSs in PSCC tissues. A) Volcano plot of the differentially expressed genes in CD74⁺ B cells from TLSs compared with those from nTLSs, with larger and redder dots indicating higher expression levels. B) Spatial in situ expression of H2BC5 , HLA‐DRA , IGKC , CYBA , ATP2A3 , and MS4A1 , with deeper colors representing higher expression levels; dashed circles denote TLS regions. C) Bubble plot showing the results of the GO enrichment analysis of genes upregulated in CD74⁺ B cells within TLSs compared with those in nTLS regions. D) Transcription factors exhibiting greater activity in CD74⁺ B cells from TLSs than in B cells from nTLSs. E) Workflow of the use of serial tissue sections from the same PSCC patient sample. The first section was used to localize CD74⁺ B cells within TLS regions, whereas the remaining two sections were used to assess the expression of key transcription factors, such as IRF4 , JUN , NFATC1 , and STAT5A , within the same TLS‐localized CD74⁺ B‐cell populations. F) Representative image of mIF staining of PSCC tissue sections from three patients. Left panel: Section was stained with CD74 (green) and CD20 (orange) to localize CD74⁺ B cells within TLS regions. Sections and show the expression of IRF4 (red), JUN (yellow), NFATC1 (purple), and STAT5A (green) at the corresponding TLS locations. Right panel: quantification of transcription factor expression levels in CD74 − B cells and CD74⁺ B cells. Two‐sided Wilcoxon test, * p < 0.05, *** p < 0.001.

Article Snippet: The antibodies used were CD20 (Proteintech, PE‐FCA65575), CD45 (Proteintech, FITC‐98117), and CD74 (Biolegend, 326812).

Techniques: Gene Expression, Functional Assay, Expressing, In Situ, Activity Assay, Staining

Journal: Advanced Science

Article Title: Spatial Transcriptional Dynamics of CD74⁺ B Cells in Tertiary Lymphoid Structures Drive Immune Evolution in Penile Squamous Cell Carcinoma

doi: 10.1002/advs.202509742

Figure Lengend Snippet: A high proportion of CD74⁺ B cells within TLSs is correlated with improved clinical prognosis in PSCC patients. A) Representative images of immunofluorescence staining for CD20 (green), CD74 (red), and DAPI (blue) in PSCC tissue sections. The area of interest (AOI) is outlined by a dashed line, scale bar = 50 µm. B) Quantitative analysis of CD74⁺CD20⁺ B‐cell coexpression in all enrolled patients ( n = 29), stratified by pathological complete response (pCR; left) and radiological treatment response (right). pCR, pathological complete response, wherein the response evaluation criteria followed RECIST 1.1; CR, complete response; PR, partial response; SD, stable disease; PD, progressive disease; RECIST, response evaluation criteria in solid tumors. C) Kaplan–Meier survival curve analysis of patients with PSCC with high ( n = 8) or low ( n = 11) CD74 expression in B cells in TLSs; p = 0.19. D) Differentiation and immune activation response patterns of CD74⁺ B cells in the TLSs of PSCC patients. Survival analysis revealed that patients with higher TLS scores had better prognoses. The differentiation of CD74⁺ B cells into various plasma cell subgroups, including IGKV4⁺, IGLV4⁺, IGHV1⁺, and IGHM⁺ plasma cells, was considered the starting point of the developmental trajectories. In the TLSs of PSCC tissues, CD74⁺ B cells activate immune responses, proliferation, and migration‐related transcription factors in naive T cells via the HLA‐DRA ligand and CD4 receptor, promoting immune activation within the tumor microenvironment. ** p < 0.01 and *** p < 0.001.

Article Snippet: The antibodies used were CD20 (Proteintech, PE‐FCA65575), CD45 (Proteintech, FITC‐98117), and CD74 (Biolegend, 326812).

Techniques: Immunofluorescence, Staining, Expressing, Activation Assay, Clinical Proteomics, Migration

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