human cd3  (Sino Biological)

Journal: Journal for Immunotherapy of Cancer

Article Title: Non-superagonist CD28-based dual-signal T cell engager targeting

doi: 10.1136/jitc-2025-013246

Figure Lengend Snippet: KK-LC-1×CD28 enhances the specific lysis of KK-LC-1-expressing target cells by T cells in vitro. ( A–C ) Dose-dependent lysis of MKN45, HGC27(oe-KK-LC-1), and HGC27 by unstimulated T cells varies proportionally with CD3 bispecific protein concentration (E:T ratio 10:1; incubation time, 48 hours). ( D ) Representative confocal microscopy images (upper) and surface projections (lower) of live/dead staining in MCSs. Viable cells (green: calcein AM) and dead cells (red: Propidium Iodide) are quantified; scale bar=100 µm. ( E ) Quantitative analysis of viability in MCSs. ( F ) Combination therapy (KK-LC-1×CD3 + KK-LC-1×CD28) induces cytotoxicity against MKN45 and HGC27(oe-KK-LC-1) cells across E:T ratios (1:1, 5:1, 10:1; 48 hours). KK-LC-1×CD3(2.5 µg/mL) KK-LC-1×CD28 (5 µg/mL). ( G ) The cytotoxic efficacy of KK-LC-1×CD3 monotherapy and combination therapy was evaluated under an E:T ratio of 10:1 for 48 hours, with or without T cell involvement. KK-LC-1×CD3 (2.5 µg/mL) KK-LC-1×CD28 (5 µg/mL). Data are represented as mean±SEM (n=3). *p<0.05; **p<0.01; ***p<0.001; ****p<0.0001; ns: not significant. E:T, effector-to-target; KK-LC-1, Kita-Kyushu Lung Cancer Antigen-1; MCSs, multicellular spheroids.

Article Snippet: Similarly, the extracellular domains of human CD3 (Sino Biological, Cat# 10977-H08H) and CD28 (Sino Biological, Cat# 90182-C08H) were individually immobilized on CM5 chips via amine coupling under identical conditions.

Techniques: Lysis, Expressing, In Vitro, Protein Concentration, Incubation, Confocal Microscopy, Staining

Journal: iScience

Article Title: Inducible deletion of Ezh2 in CD4 + T cells inhibits kidney T cell infiltration and prevents interstitial nephritis in MRL/ lpr lupus-prone mice

doi: 10.1016/j.isci.2024.111114

Figure Lengend Snippet:

Article Snippet: Then cells were blocked with anti-mouse CD16/CD32 (BD Biosciences) followed by staining with the following antibodies: anti-CD3 epsilon-VioBlue (Clone 17A2, Miltenyi Biotec), anti-TCR β-Alexa Fluor 488 (Clone H57-597, BioLegend), anti-CD4-Pacific Green (Clone RM4-5, Thermo Fisher Scientific), and anti-CD8a-PE-Cyanine5 (Clone 53-6.7, BioLegend) in PBS with 1% fetal bovine serum (FBS).

Techniques: Recombinant, Staining, Enzyme-linked Immunosorbent Assay, Diagnostic Assay, Software, RNA Sequencing

Journal: Molecular Therapy

Article Title: Engineering memory T cells as a platform for long-term enzyme replacement therapy in lysosomal storage disorders

doi: 10.1016/j.ymthe.2024.09.033

Figure Lengend Snippet: Engineered Tm cells provide IDUA systemically and reduce GAG accumulations across multiple tissues, ameliorating histopathological hallmarks of MPS I (A) Flow cytometry results of human CD45RO within organs of Tm cell-treated NSG-MPS I mice. (B) Fluorometric assay of tissue lysate IDUA activity levels at week 22 post human Tm treatment. (C) GAG content of tissue lysates at week 22 post human Tm cell treatment. (D) Urine GAG content at 0, 6, 12, and 18 weeks post Tm cell treatment. (A–D) Heterozygous (black), NSG-MPS I (red), and Tm cell-treated NSG-MPS I (blue) NSG mice ( n = 12 mice each cohort). (E) Representative histological stained images of human CD3 (top), human IDUA (middle), and LAMP-1 (bottom) of mouse cohorts. (F) Quantification of human IDUA immunohistochemistry within liver samples ( n = 4 each cohort). (G) Quantification of LAMP-1 immunohistochemistry within liver samples ( n = 4 each cohort). Statistical analysis: two-way ANOVA (A–D), one-way ANOVA (F and G) ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001.

Article Snippet: Prepared tissue sections were deparaffinized, rehydrated, and then stained with rabbit monoclonal anti-LAMP antibody (ab208943, Abcam), anti-human CD3 epsilon antibody (MAB10670, R&D Systems), or anti-human alpha-L-iduronidase antibody (AF4119, Biotechne).

Techniques: Flow Cytometry, Activity Assay, Staining, Immunohistochemistry

Journal: Molecular Therapy

Article Title: Engineering memory T cells as a platform for long-term enzyme replacement therapy in lysosomal storage disorders

doi: 10.1016/j.ymthe.2024.09.033

Figure Lengend Snippet: Engineered Tm cells have the capability to ameliorate pathological MPS I hallmarks within the central nervous system (A) Representative histological images of human CD3 staining within brain tissue. Black arrows, CD3 immunopositive cells; gray arrows, astrocytes (top). Representative histological images of human IDUA within brain tissue (middle). Representative histological images of LAMP-1 staining within brain tissue. Black arrows, neurons; gray arrows, astrocytes (bottom). (B) Quantification of LAMP-1 immunohistochemistry within brain tissue samples ( n = 4 each cohort). (C) Barnes maze time to escape of heterozygous (black), untreated NSG-MPS I (red), and Tm cell-treated NSG-MPS I mice (blue) ( n = 9). Statistical analysis: one-way ANOVA (B), two-way ANOVA (C) ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001.

Article Snippet: Prepared tissue sections were deparaffinized, rehydrated, and then stained with rabbit monoclonal anti-LAMP antibody (ab208943, Abcam), anti-human CD3 epsilon antibody (MAB10670, R&D Systems), or anti-human alpha-L-iduronidase antibody (AF4119, Biotechne).

Techniques: Staining, Immunohistochemistry