fitc anti mouse cd3  (Elabscience Biotechnology)

Journal: Molecular Therapy Oncology

Article Title: Engineered Salmonella -mediated c-di-AMP delivery activates STING to remodel the tumor microenvironment

doi: 10.1016/j.omton.2026.201185

Figure Lengend Snippet: Effect of SL disA therapy on T cells and macrophage from tumor samples (A, B) Flow Cytometry analysis of CD3 and CD8 surface markers(A) and CD3 + CD8 + cells statistical graph (B). (C, D) Flow cytometry analysis of CD3 and CD4 surface markers (C) and CD3 + CD4 + cells statistical graph (D). (E, F) Flow cytometry analysis of F4/80 and CD86 surface markers (E) and F4/80 + CD86 + cells statistical graph (F). Data are expressed as mean ± SEM, n = 3. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001, one-way ANOVA with Tukey’s multiple comparisons tests.

Article Snippet: The following antibodies were used: FITC anti-mouse F4/80 (clone CI: A3-1), APC anti-mouse CD86 (clone GL-1), FITC anti-mouse CD3 (clone 17A2), APC anti-mouse CD4 (clone GK1.5), and APC anti-mouse CD8 (clone YTS-169), all purchased from Elabscience Biotechnology Co., Ltd.

Techniques: Flow Cytometry

Journal: Frontiers in Immunology

Article Title: Translational 3D in vitro models for immunotherapy testing: from reconstituted organoid co-culture assays to autologous ex vivo patient tissues

doi: 10.3389/fimmu.2026.1721016

Figure Lengend Snippet: Co-culture assay allows versatile IO drug testing. (a) Heatmap showing reduced organoid volume upon treatment with CD3-EGFR (dosed between 2000 and 0.20 ng/ml) compared to the isotype control within eight organoid models. Representative images of two cervical models are presented, showing effect of isotype and CD3-EGFR exposure. Bar graphs show image quantification results: normalized organoid volume and IFN-γ levels measured in the supernatant of the co-cultures. (b) Organoid model CV19046B was co-cultured with EGFR targeting and non-transduced CAR T cells showing images, immune cell infiltration in the organoids, total organoid volume and cytokine levels of IFN-γ and IL-12p70 measured in the supernatants. (c) Luminescence of lung organoid model LU5178-Lucg co-cultured with pre-activated T cells upon treatment with Pembrolizumab or the isotype control. Set of n=4 technical replicates with mean and SD shown. (d) Left: Claudin 18.2 ( CLDN18 ) gene expression in four Pancreatic cancer (PA) models as quantified by bulk RNAseq analysis. Right: Claudin 18.2 protein expression determined by FACS analysis. Per condition, n=1; minimum of 10.000 events counted per condition). (e) PA organoid models were co-cultured with PBMCs and treated with 0.001-10 µg/mL AMG910 or 10 µg hIgG control. Graphs show the percentage of CD69 + PBMCs (left), PBMC infiltration in the organoids (middle) and total organoid volume normalized to the hIgG control (right). Statistical analysis used for 3a, 3e is ordinary One-way ANOVA, followed by post-hoc multiple comparison test using Tukey’s methodology. For 3b means are compared using unpaired t-test. For 3c a Two-Way ANOVA was used, followed by multiple comparisons using Dunnett methodology. ns, not significant. Scale bars in 3a and 3b represent 500 µm.

Article Snippet: Test treatments included bispecific antibodies targeting EGFR and CD3 (0.2 ng/mL-2 μg/ml) (Creative Biolabs- #CBL-BilE003-11) or BSA and CD3 control (2 μg/ml) (Creative Biolabs, Cat #BSFV-C001), Claudin 18.2 targeting BiTE AMG910 (1 ng/mL-10 μg/mL) (MCE, Cat #HY-P99350) or Ultra-LEAFTM Purified Human IgG1 Isotype Ctrl (10 μg/mL) (Biolegend, Cat #403501), EGFRscFv-CD28-CD3ζ CAR T-cells and Non transduced T cells (both ProMab Biotechnologies).

Techniques: Co-culture Assay, Control, Cell Culture, Gene Expression, RNA sequencing, Expressing, Comparison

Journal: Frontiers in Immunology

Article Title: Translational 3D in vitro models for immunotherapy testing: from reconstituted organoid co-culture assays to autologous ex vivo patient tissues

doi: 10.3389/fimmu.2026.1721016

Figure Lengend Snippet: ICI testing in Ex Vivo Patient Tissue assay in native TME environment. (a) Left panel shows cropped images from sample OLS-016 shown as a projection from an image stack, part of a single well. Raw images of nuclei stained with Hoechst33342 and actin cytoskeleton with rhodamine-Phalloidin. Object segmentation output after Ominer processing, separated for large object and single cells. Middle and right panels show high resolution images of samples OLP-028 and OLS-147 immuno-stained with EpCAM and CD3, and CD45 antibodies, confirming presence of both multicellular and single tumor cells as well as immune infiltrated cells in the resection samples present after 6 days culture. (b) Quantified HCI readout of untreated negative control (medium), immuno-stimulatory controls CD3/CD28 Dynabeads and SEA, immune checkpoint inhibiting (ICI) Pembrolizumab and Ipilimumab, and toxicity control Staurosporin in NSCLC samples OLS-016, OLS-113, OLS-090. Normalized, as percentage of control (poc), tumoroid size (median) shown in a violin plot, object count (median) shown in a box plot, single cell count (mean) shown in a bar chart. n=8 replicates per condition (c) Cytokine analyses in pooled supernatants per treatment condition collected from OLS-016 assay plate for analytes IFN-γ, Granzyme B, and IL-2. Averages of two pooled sets of quadruplicates with standard deviation shown, no statistics shown. (d) Single DAB staining and Multiplex IF (mIF) on FFPE embedded tumor fragments from patient-derived NSCLC samples. Single staining for epithelial marker Pan-CK, TME and immune cell and biomarkers α-SMA, CD45, CD3, CD4, CD8, CD68, CD80, CD163, FoxP3, PD-L1, PD1, CTLA-4. mIF for epithelial marker EpCAM, and TME markers α-SMA and CD45. Quantification on the stacked bar shows the representative presence of each subpopulation in the mIF sample. (e) Flow cytometry analyses of the immune populations (CD45 + ) of sample OLS-090 identifies most cells as basic T cells (CD3 + ) next to myeloid cells (CD11b + ) presented in a donut plot. (f) A more extensive flow cytometry panel tracks subsets of immune cells at baseline and timepoints 24h and 144h represented in donut plots. Overall, relative proportions of cells remain stable in the first day, whereas after 144h fewer cells could be recovered from the sample, of which the CD8 + population was progressively less present. (g) Heatmaps of n=49 NSCLC, n=6 Ovarian cancer, n=10 Breast cancer, and n=5 Bladder cancer ex vivo samples’ responses to controls and immuno-stimulatory and ICI compounds in EVPT platform. Total tumoroid area was normalized as % of medium control for every sample. Shown is the median value from 8 technical replicate wells. Statistical analysis used for 4b is Ordinary One-way ANOVA, followed by post-hoc multiple comparison test using Tukey’s methodology. Scale bar in 4a segmentations (left) represents 200 µm in IF 100 µm (middle, right), in 4d 100 µm.

Article Snippet: Test treatments included bispecific antibodies targeting EGFR and CD3 (0.2 ng/mL-2 μg/ml) (Creative Biolabs- #CBL-BilE003-11) or BSA and CD3 control (2 μg/ml) (Creative Biolabs, Cat #BSFV-C001), Claudin 18.2 targeting BiTE AMG910 (1 ng/mL-10 μg/mL) (MCE, Cat #HY-P99350) or Ultra-LEAFTM Purified Human IgG1 Isotype Ctrl (10 μg/mL) (Biolegend, Cat #403501), EGFRscFv-CD28-CD3ζ CAR T-cells and Non transduced T cells (both ProMab Biotechnologies).

Techniques: Ex Vivo, Staining, Negative Control, Control, Single Cell, Standard Deviation, Multiplex Assay, Derivative Assay, Marker, Flow Cytometry, Comparison

Journal: Frontiers in Immunology

Article Title: Translational 3D in vitro models for immunotherapy testing: from reconstituted organoid co-culture assays to autologous ex vivo patient tissues

doi: 10.3389/fimmu.2026.1721016

Figure Lengend Snippet: Ex vivo patient tissue assay allows IO drug efficacy assessment in native TME environment. (a) Heatmap of three NSCLC samples treated with T cell engager bi-specific antibody targeting CD3 and EGFR, or BSA in the control, Cetuximab, SEA, measured either with HCI analyses, or IFN-γ as a fast-readout for immune cell activation. (b) HCI measurements as described in (4B). (c) Heatmap of normalized tumoroid area measurements from HCI 10 primary samples embedded together with autologous PBMCs and exposed to medium, SEA, Pembrolizumab (ICI), experimental antibody compound X in low, intermediate (int), and high dose, and combination of the these with Pembrolizumab. (d) HCI measurements as described in (4b), and normalized infiltration measurement of autologous PBMCs into tumoroids. (e) Representative high-resolution images (cropped) of stained tumoroid objects (as in a) together with cell tracker labeled PBMCs. Set of n=8 technical replicates with mean or median and SD shown. Statistical analysis used for 5b, 5d is Ordinary One-way ANOVA, followed by post-hoc multiple comparison test using Tukey’s methodology. ns, not significant. Scale bar represents 50 µm.

Article Snippet: Test treatments included bispecific antibodies targeting EGFR and CD3 (0.2 ng/mL-2 μg/ml) (Creative Biolabs- #CBL-BilE003-11) or BSA and CD3 control (2 μg/ml) (Creative Biolabs, Cat #BSFV-C001), Claudin 18.2 targeting BiTE AMG910 (1 ng/mL-10 μg/mL) (MCE, Cat #HY-P99350) or Ultra-LEAFTM Purified Human IgG1 Isotype Ctrl (10 μg/mL) (Biolegend, Cat #403501), EGFRscFv-CD28-CD3ζ CAR T-cells and Non transduced T cells (both ProMab Biotechnologies).

Techniques: Ex Vivo, Control, Activation Assay, Staining, Labeling, Comparison