human ifnγ  (R&D Systems)

Journal: iScience

Article Title: Targeting ROR1 with humanized antibody drug conjugates and cytokine fusion proteins for cancer therapy

doi: 10.1016/j.isci.2026.115578

Figure Lengend Snippet: Generation of high-affinity antibodies against human ROR1 (A) The binding affinity of ROR1-targeted antibodies generated by the hybridoma technique was analyzed by ELISA against purified ROR1 extracellular domain proteins (ROR1-ECD) (replicate = 2). Amino acid sequences of variable regions were presented in . (B) Flow cytometry analysis of the binding between ROR1-targeted mouse antibodies and ROR1 presented on MDA-MB-231 cells (replicate = 2). (C) The antibodies’ isotypes and binding affinity measured by ELISA and FACS were summarized (data were represented as mean). (D) The binding affinity of chimeric antibodies was detected by ELISA against purified ROR1-ECD proteins (replicate = 2). (E) Flow cytometry analysis of the binding between chimeric antibodies and ROR1 presented on MDA-MB-231 cells (replicate = 2). The analysis of the binding on NCI-H226 cells was presented in . (F) The chimeric antibodies’ binding affinity measured by ELISA and FACS were summarized (data were represented as mean).

Article Snippet: Human IFN-gamma ELISA Kit , MULTI SCIENCES , Cat#EK180.

Techniques: Binding Assay, Generated, Enzyme-linked Immunosorbent Assay, Purification, Flow Cytometry

Journal: iScience

Article Title: Targeting ROR1 with humanized antibody drug conjugates and cytokine fusion proteins for cancer therapy

doi: 10.1016/j.isci.2026.115578

Figure Lengend Snippet: Epitope analysis of ROR1 antibodies (A) The binding affinity of antibodies against truncated ROR1 variants expressed on CHO cells was detected by FACS (replicate = 2). (B) ELISA analysis of the antibodies’ binding activity to truncated ROR1 proteins coated on ELISA plates (replicate = 2). (C) The degradation of ROR1 protein after 5 μg/mL R-001c ∼ R-009c treatment of MDA-MB-231 cells for 18 h. (D) ROR1 internalization was monitored by immunofluorescence staining and confocal microscopy. Scale bars, 20 μm. (E) The GTP-Rac1 signaling pathway was detected by western blot after treatment with R-001c, R-002c, R-004c, R-005c, and UC-961 for 18 h, followed by Wnt5a simulation for 5 min.

Article Snippet: Human IFN-gamma ELISA Kit , MULTI SCIENCES , Cat#EK180.

Techniques: Binding Assay, Enzyme-linked Immunosorbent Assay, Activity Assay, Immunofluorescence, Staining, Confocal Microscopy, Western Blot

Journal: iScience

Article Title: Targeting ROR1 with humanized antibody drug conjugates and cytokine fusion proteins for cancer therapy

doi: 10.1016/j.isci.2026.115578

Figure Lengend Snippet: Development and in vitro characterization of ROR1-humanized antibodies (A) The binding affinity of humanized antibodies was evaluated by ELISA against ROR1-ECD proteins (replicate = 2, data were represented as mean). Amino acid sequences of variable regions were presented in . (B) Flow cytometry analysis of the binding between ROR1-targeted humanized antibodies and ROR1 presented on MDA-MB-231 cells (replicate = 2, data were represented as mean). The analysis of the binding on NCI-H226 cells was presented in . (C) ROR1 internalization induced by Hu001-2 and Hu005-46 was detected by confocal microscopy. Scale bars, 50 μm. The ROR1 internalization induced by UC-961 was presented in A. (D) The internalization kinetics and binding affinity of humanized antibodies were monitored over 4 h by FACS. (E) The binding K D of Hu001-2 and Hu005-46 was evaluated by surface plasmon resonance (SPR) analysis. The binding K D of UC-961 was presented in B.

Article Snippet: Human IFN-gamma ELISA Kit , MULTI SCIENCES , Cat#EK180.

Techniques: In Vitro, Binding Assay, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Confocal Microscopy, SPR Assay

Journal: iScience

Article Title: Targeting ROR1 with humanized antibody drug conjugates and cytokine fusion proteins for cancer therapy

doi: 10.1016/j.isci.2026.115578

Figure Lengend Snippet: Development of immune-activating antibody-cytokine fusion proteins (A) Schematic diagram of the Hu001-2-IL15Rα-IL15 fusion protein. Purification and characterization of fusion proteins were presented in . (B) Binding affinity to ROR1-ECD and IL-15Rβ was detected by ELISA (replicate = 2, data were represented as mean). The binding affinity with ROR1 on MDA-MB-231 cells was presented in . (C) After 6-day culture with Hu001-2-IL15Rα-IL15, CD8 + T cell and CD56 + NK cell populations in PBMCs were analyzed by FACS. (D) IFN-γ and TNF-α secretion by PBMCs were measured by ELISA. (E) The cytotoxicity activity of fusion proteins was evaluated by co-culture of PBMCs with MDA-MB-231 cells at an effector-to-target ratio of 10:1 and 20:1 for 24 h. Results were expressed as means ± SD ( n = 3). Statistical analysis was performed using two-tailed unpaired t test with Welch’s correction, ∗∗ p < 0.01 and ∗∗∗ p < 0.001.

Article Snippet: Human IFN-gamma ELISA Kit , MULTI SCIENCES , Cat#EK180.

Techniques: Protein Purification, Binding Assay, Enzyme-linked Immunosorbent Assay, Activity Assay, Co-Culture Assay, Two Tailed Test

Journal: Bioactive Materials

Article Title: Splenic dendritic cell-targeting mRNA transfection of H-type ionizable lipid-based LNPs for enhancing tumor immunotherapy

doi: 10.1016/j.bioactmat.2026.02.018

Figure Lengend Snippet: In vivo DC maturation and T cell activation in C57BL/6J mice induced by mOVA/H 18 NPs through intravenous injection. C57BL/6J mice were vaccinated with different formulations on Day 0 and Day 5. On Day 10, the spleens of mice were collected and analyzed by flow cytometry. Quantification analysis of (A) CD80 + CD86 + DCs and (B) CD40 + DCs in the spleen. Quantification analysis of (C) CD3 + CD4 + T cells and (D) CD3 + CD8 + T cells in the spleen. (E) Quantification analysis and (F) representative flow cytometry contour plots of OVA-specific CD8 + T cells among all cell populations in the spleen. (G) Quantification analysis and (H) representative flow cytometry contour plots of IFN-γ + CD8 + T cells among all cell populations in the spleen. (I) Quantification results and (J) representative images of IFN- γ -secreting immune cells in the spleen of mice analyzed by enzyme-linked immunospot (ELISpot) assay. Data were shown as mean ± SD (n = 3).

Article Snippet: Quantification analysis of (C) CD3 + CD4 + T cells and (D) CD3 + CD8 + T cells in the spleen. (E) Quantification analysis and (F) representative flow cytometry contour plots of OVA-specific CD8 + T cells among all cell populations in the spleen. (G) Quantification analysis and (H) representative flow cytometry contour plots of IFN-γ + CD8 + T cells among all cell populations in the spleen. (I) Quantification results and (J) representative images of IFN- γ -secreting immune cells in the spleen of mice analyzed by enzyme-linked immunospot (ELISpot) assay.

Techniques: In Vivo, Activation Assay, Injection, Flow Cytometry, Enzyme-linked Immunospot

Journal: Molecular Therapy Oncology

Article Title: AAV immuno-gene therapy platform delivering vectorized cytokines defines a new modality for high-grade glioma treatment

doi: 10.1016/j.omton.2026.201183

Figure Lengend Snippet: Vectorized IFNβ drives durable signaling and complete tumor regression in human glioblastoma models in vivo (A) Sustained hIFNβ secretion in human GBM6 cells treated with AAV9-hIFNβ (red, MOI = 4E5 vg/cell) or recombinant hIFNβ cytokine (r-hIFNβ, purple, 47 IU/mL, equivalent to 114 pg/mL), measured by ELISA at indicated time points. 50% media washouts every 5 h for the first 20 h in the r-hIFNβ condition mimic in vivo cytokine clearance (half-life = 4–5 h). Full media exchanges were performed at 24, 48, 72, and 96 h post-treatment. (B) Number of differentially expressed genes (DEGs, p -Adj<0.01) in GBM6 cells 24–96 h post-treatment with AAV9-hIFNβ or r-hIFNβ vs. media controls. (C) Enrichment scores for type I IFN and TNFα response pathways across treatments and time points. (D) Heatmap of the top 10 IFN and TNFα response genes (Log2FC vs. media controls) in GBM6 cells treated as in (A). (E) Schematic of orthotopic PDX (SF11411) and cell line-derived xenograft ([CDX], GBM6-FLuc) studies in athymic nu/nu mice treated intratumorally with saline, AAV9-GFP, or AAV9-hIFNβ via CED. (F) Kaplan-Meier survival curves for PDX mice treated as in (E). Saline = black, AAV9-GFP (2E11 vg/brain) = blue, AAV9-hIFNβ (2E11 vg/brain) = red. Vertical dashed line = day of treatment (day 9). p < 0.04 by log-rank (Mantel-Cox) test. n = 30 (10 per treatment arm). (G) Longitudinal BLI of GBM6-FLuc tumor growth in CDX mice treated as in (E). Saline = black, AAV9-GFP (2E11 vg/brain) = blue, AAV9-hIFNβ (2E11 vg/brain) = red. Thin lines = individual mice, thick lines = geometric mean. Vertical dashed line = day of treatment (day 9). ∗ p < 0.04 by Kruskal-Wallis test with Dunn’s multiple comparisons correction on day 22. n = 30 (10 per treatment arm). (G′) Representative BLI images from each treatment group 11 days post-treatment. (H) Kaplan-Meier survival curves for CDX mice. p < 0.001 by log-rank (Mantel-Cox) test. (I) Distribution of treatment responses in CDX by BLI flux (photons/second) at day 27. Tumor free = BLI flux <2.5 × 10 5 p/s, tumor reduction = ≥30% decrease from assignment on day 9, no change = between 30% decrease and 20% increase from assignment on day 9, tumor growth = ≥20% increase from assignment on day 9, death = mice that died before day 27. (J) Dose-response analysis of AAV9-hIFNβ efficacy in CDX mice. AAV9-GFP (2E11 vg/brain) = blue, AAV9-hIFNβ hi (2E11 vg/brain) = solid red, and AAV9-hIFNβ lo (1E11 vg/brain) = dashed red. Thin lines = individual mice, thick lines = geometric mean. Vertical dashed line = day of treatment (day 9). ∗∗ p < 0.02 by Kruskal-Wallis test with Dunn’s multiple comparisons correction on day 20. n = 45 (15 per treatment arm). For data interpretation, tumor burden threshold = 2.5 × 10 5 . (J′) Representative BLI images of tumors 11 days post-treatment. (K) Kaplan-Meier survival curves from (J). p < 0.002 (AAV9-hIFNβ hi), p < 0.005 (AAV9-hIFNβ lo) by log-rank (Mantel-Cox) test compared to AAV9-GFP. (I) Distribution of treatment responses in CDX mice at day 27 by BLI flux as in (I).

Article Snippet: 24, 48, 72, and 96 h after treatment, cell supernatants were collected and IFN variant levels were measured using IFN ELISA kits following the manufacturer’s instructions (human IFNα [PBL Cat# 41135-1], human IFNβ [PBL Cat#41410], and human IFNγ [R&D Systems Cat#: DIF50C]).

Techniques: In Vivo, Recombinant, Enzyme-linked Immunosorbent Assay, Derivative Assay, Saline

Journal: Molecular Therapy Oncology

Article Title: AAV immuno-gene therapy platform delivering vectorized cytokines defines a new modality for high-grade glioma treatment

doi: 10.1016/j.omton.2026.201183

Figure Lengend Snippet: Spatial transcriptomics reveals rapid, localized transcriptional remodeling of the tumor microenvironment following vectorized hIFNβ treatment (A) Coronal brain sections from representative human GBM6-FLuc CDX mice collected pre-treatment (0 h, n = 1) or 48 h ( n = 1) after intratumoral AAV9-hIFNβ infusion (2E11 vg/brain), stained with H&E (left) and subjected to Visium Spatial Gene Expression profiling (right). Annotated clusters were assigned based on anatomical localization and marker gene expression. Dashed lines denote tumor borders. Scale bars, 1 mm. (B) Top 10 marker genes for each spatially resolved cluster identified across 0 h and 48 h datasets. Values are shown as log-normalized expression centered at 0 (Seurat “scale.data”). (C) Spatial expression of canonical human GBM tumor markers ( CD44 , VIM , TOP2A , and NOTCH1 ) delineating tumor and peri-tumor regions before (0 h) (top) and after (bottom) (48 h) AAV9-hIFNβ treatment. (D) Expression maps of the human IFNβ payload and hallmark IFN-response genes ( CXCL10 , IFIT1 , and IFIT2 ), demonstrating tumor-restricted transgene expression and induction of an IFN-specific transcriptional program within 48 h. (E) Spatial expression of host mouse immune-response genes ( Gfap , Ifitm3 , and Irf7 ) showing localized activation of astroglial and innate immune pathways proximal to the tumor. (F) Integrated datasets (0 and 48 h) visualized using canonical correlation analysis (CCA), showing distinct clustering of tumor and peri-tumor regions (left) and enrichment of IFN-response gene module expression (right). (G) Volcano plot depicting differential gene expression between 0- and 48-h tumor clusters. Red, IFN-response genes; gray, other significantly upregulated genes ( p -Adj <0.01); blue, non-significant. (H) Top enriched Gene Ontology (GO) terms among upregulated genes in 48-h tumor cells, highlighting interferon and inflammatory response pathways (∗∗ p -Adj <0.01; ∗ p -Adj <0.05).

Article Snippet: 24, 48, 72, and 96 h after treatment, cell supernatants were collected and IFN variant levels were measured using IFN ELISA kits following the manufacturer’s instructions (human IFNα [PBL Cat# 41135-1], human IFNβ [PBL Cat#41410], and human IFNγ [R&D Systems Cat#: DIF50C]).

Techniques: Spatial Transcriptomics, Staining, Gene Expression, Marker, Expressing, Activation Assay

Journal: Molecular Medicine Reports

Article Title: Elevated IgG levels induce an M2-to-M1 phenotypic shift in mucosal macrophages and restrict the growth of invasive sphenoid sinus pituitary adenomas

doi: 10.3892/mmr.2026.13878

Figure Lengend Snippet: IFN-γ suppresses tumor growth and invasion. (A) Cytokine profiling of co-culture supernatants via ELISAs: IFN-γ, IL-1β, IL-6, IL-10, TGF-β and TNF-α. (B-D) Spatial expression patterns of IFN-γ. (B) Immunofluorescence imaging of the invasive front in SSIT, showing DAPI (blue), IBA-1 + macrophages (red) and IFN-γ + signals (green), and grayscale intensity distribution. (C) Immunofluorescence imaging of TIM and NIM, showing DAPI (blue), IBA-1 + macrophages (red) and IFN-γ + signals (green). (D) Quantification of relative IFN-γ expression in TIM and NIM. (E) Representative Ki-67 immunohistochemistry images of SSIT cases stratified into IFN-γ-high and IFN-γ-low groups (n=5 each; median split). (F) Quantification of Ki-67 index comparing the two groups. (G) EdU staining demonstrating dose-dependent suppression of TtT/GF pituitary adenoma cell proliferation by IFN-γ (0–100 ng/ml; 48 h). (H) Representative flow cytometry histograms for cell cycle analysis of cells treated with IFN-γ (0–100 ng/ml) in the absence (0 µM) or presence (5 µM) of ruxolitinib. (I) Stacked bar plot showing the percentages of cells in the G 1 , S and G 2 /M phases under the same treatment conditions. (A) One-way ANOVA with Tukey's post hoc multiple comparisons test. (D and F) Unpaired two-tailed Student's t-test. *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001. CTRL, control; DMC, digested mucosal culture; EdU, 5-ethynyl-2′-deoxyuridine; IBA-1, ionised calcium binding adaptor molecule 1; MTC, mucosal tissue culture; NIM, non-invaded mucosa; ns, not significant; PE-A, phycoerythrin-area; SSIT, sphenoid sinus-invasive tumor; TIM, tumor-invaded mucosa.

Article Snippet: Human IFN-γ, IL-1β, IL-6, IL-10, TGF-β and TNF-α levels were quantified using commercial ELISA kits (Human IFN-γ ELISA kit, cat. no. E-EL-H0108; Human IL-1β ELISA kit, cat. no. E-EL-H0149; Human IL-6 ELISA kit, cat. no. E-EL-H0102; Human IL-10 ELISA kit, cat. no. E-EL-H0103; Human TGF-β ELISA kit, cat. no. E-EL-H0110; Human TNF-α ELISA kit, cat. no. E-EL-H0109; Wuhan Elabscience Biotechnology Co., Ltd.).

Techniques: Co-Culture Assay, Expressing, Immunofluorescence, Imaging, Immunohistochemistry, Staining, Flow Cytometry, Cell Cycle Assay, Two Tailed Test, Control, Binding Assay

Journal: Molecular Medicine Reports

Article Title: Elevated IgG levels induce an M2-to-M1 phenotypic shift in mucosal macrophages and restrict the growth of invasive sphenoid sinus pituitary adenomas

doi: 10.3892/mmr.2026.13878

Figure Lengend Snippet: Elevated IgG levels drive macrophage M2-to-M1 reprogramming. (A) Sphenoid sinus-invasive tumor cases stratified into CD19-high (n=5) and CD19-low (n=5) groups based on the cohort median of CD19 + B cell density, with (B) quantitative analyses of macrophage polarization (M1-like versus M2-like). (C) Dural-invasive tumor and non-invasive tumor cases stratified into IgG-high (n=27) and IgG-low (n=26) groups based on the cohort median of relative IgG immunohistochemistry staining intensity, with (D) quantitative analyses of M1-like/M2-like macrophage proportions. (E and F) RAW264.7 macrophages were pre-polarized with IL-4 (20 ng/ml) or with lipopolysaccharide (100 ng/ml) plus IFN-γ (20 ng/ml) for 24 h, followed by IgG (10 µg/ml) exposure. Relative (E) IL-6 and (F) TNF-α mRNA expression in RAW264.7 macrophages pre-polarized to M0, M1 or M2 states. (G) Representative flow cytometric cell-cycle profiles of TtT/GF cells following the indicated treatments. (H) Stacked bar plot summarizing the percentages of cells from (G) in G 1 , S and G 2 /M phases. (I) Representative images from the scratch wound assay at 0, 24, 48 and 72 h under the indicated treatments. (J) Quantification of scratch wound closure. (K) Representative western blot images showing total STAT1, p-STAT1, total STAT3, p-STAT3 and β-actin levels in cells treated with IFN-γ (100 ng/ml), IL-6 (100 ng/ml), IFN-γ + IL-6 (50 ng/ml each), ruxolitinib (5 µM) or IFN-γ + IL-6 (50 ng/ml each) plus ruxolitinib (5 µM), as indicated. (L) Densitometric semi-quantification of p-STAT1/STAT1 (ratio). (B and D) Unpaired two-tailed Student's t-test. (E, F, J and L) One-way ANOVA with Tukey's post hoc multiple comparisons test. *P<0.05, ***P<0.001, ****P<0.0001. CTRL, control; IBA-1, ionised calcium binding adaptor molecule 1; ns, not significant; p-, phosphorylated; PE-A, phycoerythrin-area.

Article Snippet: Human IFN-γ, IL-1β, IL-6, IL-10, TGF-β and TNF-α levels were quantified using commercial ELISA kits (Human IFN-γ ELISA kit, cat. no. E-EL-H0108; Human IL-1β ELISA kit, cat. no. E-EL-H0149; Human IL-6 ELISA kit, cat. no. E-EL-H0102; Human IL-10 ELISA kit, cat. no. E-EL-H0103; Human TGF-β ELISA kit, cat. no. E-EL-H0110; Human TNF-α ELISA kit, cat. no. E-EL-H0109; Wuhan Elabscience Biotechnology Co., Ltd.).

Techniques: Immunohistochemistry, Staining, Expressing, Scratch Wound Assay Assay, Western Blot, Two Tailed Test, Control, Binding Assay

Journal: Molecular Medicine Reports

Article Title: Elevated IgG levels induce an M2-to-M1 phenotypic shift in mucosal macrophages and restrict the growth of invasive sphenoid sinus pituitary adenomas

doi: 10.3892/mmr.2026.13878

Figure Lengend Snippet: Anti-CD47 mAb enhances ADCP to suppress tumor cell proliferation. (A) Immunofluorescence staining of CD47 (red) and DAPI (blue) in a representative subset of non-invasive tumor, dural-invasive tumor and sphenoid sinus-invasive tumor cases (n=10 per group). (B) Paired comparison of CD47 fluorescence intensity at the IF versus the TC. (C) RAW264.7 macrophages were pre-polarized with IL-4 (20 ng/ml) or with lipopolysaccharide (100 ng/ml) plus IFN-γ (20 ng/ml) for 24 h, followed by anti-CD47 mAb (10 µg/ml) treatment for 12 h. Quantitative PCR was used to analyze polarization/activation markers. (D) Schematic illustrating anti-CD47 mAb-mediated blockade of the CD47-SIRPα axis and enhancement of ADCP. (E) EdU assay of TtT/GF cell proliferation in a Transwell co-culture with anti-CD47 mAb-treated polarized macrophages. (F) Quantification of EdU-positive cells. (G) Representative microscopy images and flow cytometry plots showing macrophage phagocytosis of pHrodo™ Red-labeled GFP-TtT/GF cells. (H) Quantification of phagocytosis (%). (B) Paired two-tailed Student's t-test. (C, F and H) One-way ANOVA with Tukey's post hoc multiple comparisons test. **P<0.01, ***P<0.001, ****P<0.0001. ADCP, antibody-dependent cellular phagocytosis; Arg-1, arginase 1; EdU, 5-ethynyl-2′-deoxyuridine; FcγR, Fcγ receptor; GFP, green fluorescent protein; IF, invasive front; mAb, monoclonal antibody; NOS2, nitric oxide synthase 2; ns, not significant; PE, phycoerythrin; SIRPα, signal regulatory protein-α; SSCA, side scatter area; TC, tumor core.

Article Snippet: Human IFN-γ, IL-1β, IL-6, IL-10, TGF-β and TNF-α levels were quantified using commercial ELISA kits (Human IFN-γ ELISA kit, cat. no. E-EL-H0108; Human IL-1β ELISA kit, cat. no. E-EL-H0149; Human IL-6 ELISA kit, cat. no. E-EL-H0102; Human IL-10 ELISA kit, cat. no. E-EL-H0103; Human TGF-β ELISA kit, cat. no. E-EL-H0110; Human TNF-α ELISA kit, cat. no. E-EL-H0109; Wuhan Elabscience Biotechnology Co., Ltd.).

Techniques: Immunofluorescence, Staining, Comparison, Fluorescence, Real-time Polymerase Chain Reaction, Activation Assay, EdU Assay, Co-Culture Assay, Microscopy, Flow Cytometry, Labeling, Two Tailed Test

Journal: Molecular Medicine Reports

Article Title: Elevated IgG levels induce an M2-to-M1 phenotypic shift in mucosal macrophages and restrict the growth of invasive sphenoid sinus pituitary adenomas

doi: 10.3892/mmr.2026.13878

Figure Lengend Snippet: Summary graphic illustration. This illustration summarizes the proposed model during pituitary adenoma invasion. The tumor invasive front abuts an intact sphenoid sinus mucosa, forming a distinct boundary. The mucosal compartment is enriched for ionised calcium binding adaptor molecule 1-positive macrophages with an M1-like predominance and IgG-high B cells. B cell-derived IgG promotes M2-to-M1 macrophage reprogramming, while coordinated IFN-γ and IL-6 production establishes a tumor-suppressive cytokine gradient that decreases from mucosa toward the tumor core, constraining proliferation and migration via JAK-STAT1 activation. Therapeutically, anti-CD47 monoclonal antibody blocks the CD47-SIRPα ‘don't-eat-me’ axis and augments antibody-dependent cellular phagocytosis, highlighting a strategy for immune checkpoint-targeted therapy that may complement surgical management. FcR, Fc receptor; JAK, Janus kinase; mAb, monoclonal antibody; p-, phosphorylated; SIRPα, signal regulatory protein-α.

Article Snippet: Human IFN-γ, IL-1β, IL-6, IL-10, TGF-β and TNF-α levels were quantified using commercial ELISA kits (Human IFN-γ ELISA kit, cat. no. E-EL-H0108; Human IL-1β ELISA kit, cat. no. E-EL-H0149; Human IL-6 ELISA kit, cat. no. E-EL-H0102; Human IL-10 ELISA kit, cat. no. E-EL-H0103; Human TGF-β ELISA kit, cat. no. E-EL-H0110; Human TNF-α ELISA kit, cat. no. E-EL-H0109; Wuhan Elabscience Biotechnology Co., Ltd.).

Techniques: Binding Assay, Derivative Assay, Migration, Activation Assay

Journal: Human Mutation

Article Title: Whole‐Exome Sequencing to Screen Personal Neoantigens With High Immunogenicity in Patients With Microsatellite Stability (MSS)–Advanced Colorectal Cancer

doi: 10.1155/humu/3876230

Figure Lengend Snippet: Evaluating the immunogenicity of neoantigens from patients with CRC. Autologous PBMCs were stimulated with candidate mutated peptides every 3 days in the presence of IL‐2. On Day 10, T‐cell responses to each antigen were measured by an IFN‐ γ ELISpot assay. The PBMCs were obtained from P1 with CRC. No peptide (medium only) or VSV‐NP 43-69 (STKVALNDLRAYVYQGIKSGNPSILHI) stimulation was used as a control. Data are presented as mean ± SD of three independent experiments. ∗ p < 0.05 and ∗∗ p < 0.01 compared with IFN‐ γ production by PBMCs stimulated without peptide or with VSV‐NP 43-69 .

Article Snippet: After 10 days, the IFN‐ γ response from the prestimulated T cells against neoantigens was assessed using ELISpot assays with a Human IFN‐ γ ELISpot kit (Mabtech).

Techniques: Immunopeptidomics, Enzyme-linked Immunospot, Control

Journal: Human Mutation

Article Title: Whole‐Exome Sequencing to Screen Personal Neoantigens With High Immunogenicity in Patients With Microsatellite Stability (MSS)–Advanced Colorectal Cancer

doi: 10.1155/humu/3876230

Figure Lengend Snippet: Cytotoxicity of NRTs raised by in vitro stimulation of PBLs of P1. NRTs were induced with autologous ZNF169‐A275S– and CDH4‐V456M–pulsed DCs derived from the PBLs of P1. On Day 7, after the third stimulation, the NRTs were harvested for analysis. (A) IFN‐ γ secretion by NRT lines in response to mutated and WT peptides. IFN‐ γ ‐positive SFCs/10 5 NRTs were detected by cytokine‐specific ELISpot. (B–F) Cytotoxicity at the indicated E:T ratios measured by a CCK‐8 kit. Peptide‐specific targets were mutated protein–pulsed T2 cells and minimally nucleated SW480 cells, whereas VSV‐NP 43-69 –pulsed T2 cells, T2 cells alone, and SW480 cells alone were used as controls. Data are expressed as mean ± SEM ( ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001).

Article Snippet: After 10 days, the IFN‐ γ response from the prestimulated T cells against neoantigens was assessed using ELISpot assays with a Human IFN‐ γ ELISpot kit (Mabtech).

Techniques: In Vitro, Derivative Assay, Enzyme-linked Immunospot, CCK-8 Assay

Journal: Microbiology Spectrum

Article Title: MPT64 antigen-induced immune responses as a novel diagnostic tool for tuberculosis

doi: 10.1128/spectrum.03381-24

Figure Lengend Snippet: IFN-γ levels produced by lymphocytes stimulated by secreted-type proteins. ( A ) ELISA analysis of IFN-γ levels in splenocytes of immunized mice at different times (* P < 0.001 compared with MPT64 group). ( B ) Identification by the SDS-PAGE of MPT64. ( C ) WB identification of supernatant versus bacterial precipitate at different time points after MPT64 induction expression.

Article Snippet: IFN-γ levels were measured using the human IFN-γ T-SPOT.TB kit (DAKEWE Biotechnology) and quantified as spot-forming units (SFUs) using an immunospot image analyzer (BioReader-E, BioSYS, Germany).

Techniques: Produced, Enzyme-linked Immunosorbent Assay, SDS Page, Expressing

Journal: Microbiology Spectrum

Article Title: MPT64 antigen-induced immune responses as a novel diagnostic tool for tuberculosis

doi: 10.1128/spectrum.03381-24

Figure Lengend Snippet: The release of IFN-γ from both MPT64 and MPT64+CLE was much higher in active pulmonary TB patients than in the healthy control population. ( A ) IFN-γ levels of active tuberculosis and healthy control MPT64. ( B ) The ROC curve of the ELISpot for MPT64. ( C ) IFN-γ levels of active tuberculosis and healthy control MPT64+CLE. ( D ) The ROC curve of the ELISpot for MPT64+CLE. ( E ) Representative plot of MPT64 protein as antigen-stimulated PBMC in TB humans and healthy controls. ( F ) Representative plot of MPT64+CLE protein as antigen-stimulated PBMC in TB humans and healthy controls. “****” indicates a statistically significant difference.

Article Snippet: IFN-γ levels were measured using the human IFN-γ T-SPOT.TB kit (DAKEWE Biotechnology) and quantified as spot-forming units (SFUs) using an immunospot image analyzer (BioReader-E, BioSYS, Germany).

Techniques: Control, Enzyme-linked Immunospot

Journal: Microbiology Spectrum

Article Title: MPT64 antigen-induced immune responses as a novel diagnostic tool for tuberculosis

doi: 10.1128/spectrum.03381-24

Figure Lengend Snippet: Dominant epitopes in the T and B cells of the MPT64 protein. ( A ) ELISA measurement of IFN-γ release of each peptide and the complete sequence of MPT64. ( B ) The ELISA measurement of IFN-γ release from TB human PBMC stimulation by each peptide and the MPT64 complete sequence. ( C ) DTH of each polypeptide and the complete sequence of MPT64 in the mouse. ( D ) ELISA absorbance of the affinity of each polypeptide and the complete sequence of MPT64 to the MPT64 rabbit polyclonal antibody. ( E ) Spot hybridization shows the affinity of each polypeptide and the complete MPT64 sequence with the MPT64 rabbit polyclonal antibody. ( F ) Schematic diagram of the 3D predicted structure of each polypeptide, as well as the complete sequence of MPT64.

Article Snippet: IFN-γ levels were measured using the human IFN-γ T-SPOT.TB kit (DAKEWE Biotechnology) and quantified as spot-forming units (SFUs) using an immunospot image analyzer (BioReader-E, BioSYS, Germany).

Techniques: Enzyme-linked Immunosorbent Assay, Sequencing, Hybridization