interferon γ  (Cusabio)

Journal: bioRxiv

Article Title: Inexpensive High-Throughput Multiplexed Cytokine Detection for Tuberculosis Diagnostics Using Amplified Enzymatic Metallization

doi: 10.64898/2026.01.27.700981

Figure Lengend Snippet: (a) Schematic of laser-cut PDMS microwell array reversibly assembled on a polystyrene petri dish and workflow of enzymatically amplified silver metallization using Biotin-BSA and poly-HRP-SA. (b) Cell-phone image of silver metallized wells after assay. (c) Biotin-BSA dilution curve obtained from cell-phone image silver darkness quantification in ImageJ. (d) Assay stack for initial biotinyl tyramide signal amplification testing using biotinylated anti-IFN-γ as capture. (e) Dilution curve of silver darkness of increasing biotinylated anti-IFN-γ as target and 10 µg/mL, 1 µg/mL, and 0 µg/mL biotinyl tyramide. All assays were repeated three (n=3) times and fit lines are four-parameter logistic sigmoidal curves.

Article Snippet: Recombinant human IFN-γ protein (11725-HNAS), biotinylated anti-human IFN-γ antibody (11725-R209-B), anti-human IFN-γ antibody (11725-R238), HRP-conjugated anti-human IFN-γ antibody (11725-R209-H), recombinant human TNF-α protein (10602-HNAE), biotinylated anti-human TNF-α antibody (10602-MM08-B), and anti-human TNF-α antibody (10602-MM01) were obtained from SinoBiological.

Techniques: Amplification

Journal: bioRxiv

Article Title: Inexpensive High-Throughput Multiplexed Cytokine Detection for Tuberculosis Diagnostics Using Amplified Enzymatic Metallization

doi: 10.64898/2026.01.27.700981

Figure Lengend Snippet: (a) Schematic of immunoassay for cytokine detection. (b) Initial LOD optimization for the IFN-γ assay. (c-e) IFN-γ (n=2), IL-2 (n=5), TNF-α (n=2) dilution curves using recombinant cytokines (f-h) Correlation plots of ELISA absorbance values at 450 nm versus silver darkness values for IFN-γ (f), TNF-α (g), and IL-2 (h). All fit lines are four-parameter logistic sigmoidal curves.

Article Snippet: Recombinant human IFN-γ protein (11725-HNAS), biotinylated anti-human IFN-γ antibody (11725-R209-B), anti-human IFN-γ antibody (11725-R238), HRP-conjugated anti-human IFN-γ antibody (11725-R209-H), recombinant human TNF-α protein (10602-HNAE), biotinylated anti-human TNF-α antibody (10602-MM08-B), and anti-human TNF-α antibody (10602-MM01) were obtained from SinoBiological.

Techniques: Recombinant, Enzyme-linked Immunosorbent Assay

Journal: bioRxiv

Article Title: Inexpensive High-Throughput Multiplexed Cytokine Detection for Tuberculosis Diagnostics Using Amplified Enzymatic Metallization

doi: 10.64898/2026.01.27.700981

Figure Lengend Snippet: (a) Schematic demonstrating the stimulation groups and controls. Dilution curves for IFN-γ (b-c), IL-2 (d-e), and TNF-α (f-g) in pooled LTB+ and LTB-clinical supernatant samples. Clinical samples consisted of NIL (negative control), Ag1, Ag2, and Mitogen (positive control) groups (n=2).

Article Snippet: Recombinant human IFN-γ protein (11725-HNAS), biotinylated anti-human IFN-γ antibody (11725-R209-B), anti-human IFN-γ antibody (11725-R238), HRP-conjugated anti-human IFN-γ antibody (11725-R209-H), recombinant human TNF-α protein (10602-HNAE), biotinylated anti-human TNF-α antibody (10602-MM08-B), and anti-human TNF-α antibody (10602-MM01) were obtained from SinoBiological.

Techniques: Negative Control, Positive Control

Journal: bioRxiv

Article Title: Inexpensive High-Throughput Multiplexed Cytokine Detection for Tuberculosis Diagnostics Using Amplified Enzymatic Metallization

doi: 10.64898/2026.01.27.700981

Figure Lengend Snippet: Individual participant silver darkness values for IFN-γ (a), IL-2 (b), and TNF-α (c) detection from NIL, Ag1, Ag2, and Mitogen samples of IGRA+ and IGRA-patients using a sample dilution of 1:10. Average of n=2 repeats was plotted for each sample.

Article Snippet: Recombinant human IFN-γ protein (11725-HNAS), biotinylated anti-human IFN-γ antibody (11725-R209-B), anti-human IFN-γ antibody (11725-R238), HRP-conjugated anti-human IFN-γ antibody (11725-R209-H), recombinant human TNF-α protein (10602-HNAE), biotinylated anti-human TNF-α antibody (10602-MM08-B), and anti-human TNF-α antibody (10602-MM01) were obtained from SinoBiological.

Techniques:

Journal: bioRxiv

Article Title: Inexpensive High-Throughput Multiplexed Cytokine Detection for Tuberculosis Diagnostics Using Amplified Enzymatic Metallization

doi: 10.64898/2026.01.27.700981

Figure Lengend Snippet: (a) Schematic of laser-cut PDMS microwell array reversibly assembled on a polystyrene petri dish and workflow of enzymatically amplified silver metallization using Biotin-BSA and poly-HRP-SA. (b) Cell-phone image of silver metallized wells after assay. (c) Biotin-BSA dilution curve obtained from cell-phone image silver darkness quantification in ImageJ. (d) Assay stack for initial biotinyl tyramide signal amplification testing using biotinylated anti-IFN-γ as capture. (e) Dilution curve of silver darkness of increasing biotinylated anti-IFN-γ as target and 10 µg/mL, 1 µg/mL, and 0 µg/mL biotinyl tyramide. All assays were repeated three (n=3) times and fit lines are four-parameter logistic sigmoidal curves.

Article Snippet: Recombinant human IFN-γ protein (11725-HNAS), biotinylated anti-human IFN-γ antibody (11725-R209-B), anti-human IFN-γ antibody (11725-R238), HRP-conjugated anti-human IFN-γ antibody (11725-R209-H), recombinant human TNF-α protein (10602-HNAE), biotinylated anti-human TNF-α antibody (10602-MM08-B), and anti-human TNF-α antibody (10602-MM01) were obtained from SinoBiological.

Techniques: Amplification

Journal: bioRxiv

Article Title: Inexpensive High-Throughput Multiplexed Cytokine Detection for Tuberculosis Diagnostics Using Amplified Enzymatic Metallization

doi: 10.64898/2026.01.27.700981

Figure Lengend Snippet: (a) Schematic of immunoassay for cytokine detection. (b) Initial LOD optimization for the IFN-γ assay. (c-e) IFN-γ (n=2), IL-2 (n=5), TNF-α (n=2) dilution curves using recombinant cytokines (f-h) Correlation plots of ELISA absorbance values at 450 nm versus silver darkness values for IFN-γ (f), TNF-α (g), and IL-2 (h). All fit lines are four-parameter logistic sigmoidal curves.

Article Snippet: Recombinant human IFN-γ protein (11725-HNAS), biotinylated anti-human IFN-γ antibody (11725-R209-B), anti-human IFN-γ antibody (11725-R238), HRP-conjugated anti-human IFN-γ antibody (11725-R209-H), recombinant human TNF-α protein (10602-HNAE), biotinylated anti-human TNF-α antibody (10602-MM08-B), and anti-human TNF-α antibody (10602-MM01) were obtained from SinoBiological.

Techniques: Recombinant, Enzyme-linked Immunosorbent Assay

Journal: bioRxiv

Article Title: Inexpensive High-Throughput Multiplexed Cytokine Detection for Tuberculosis Diagnostics Using Amplified Enzymatic Metallization

doi: 10.64898/2026.01.27.700981

Figure Lengend Snippet: (a) Schematic demonstrating the stimulation groups and controls. Dilution curves for IFN-γ (b-c), IL-2 (d-e), and TNF-α (f-g) in pooled LTB+ and LTB-clinical supernatant samples. Clinical samples consisted of NIL (negative control), Ag1, Ag2, and Mitogen (positive control) groups (n=2).

Article Snippet: Recombinant human IFN-γ protein (11725-HNAS), biotinylated anti-human IFN-γ antibody (11725-R209-B), anti-human IFN-γ antibody (11725-R238), HRP-conjugated anti-human IFN-γ antibody (11725-R209-H), recombinant human TNF-α protein (10602-HNAE), biotinylated anti-human TNF-α antibody (10602-MM08-B), and anti-human TNF-α antibody (10602-MM01) were obtained from SinoBiological.

Techniques: Negative Control, Positive Control

Journal: bioRxiv

Article Title: Inexpensive High-Throughput Multiplexed Cytokine Detection for Tuberculosis Diagnostics Using Amplified Enzymatic Metallization

doi: 10.64898/2026.01.27.700981

Figure Lengend Snippet: Individual participant silver darkness values for IFN-γ (a), IL-2 (b), and TNF-α (c) detection from NIL, Ag1, Ag2, and Mitogen samples of IGRA+ and IGRA-patients using a sample dilution of 1:10. Average of n=2 repeats was plotted for each sample.

Article Snippet: Recombinant human IFN-γ protein (11725-HNAS), biotinylated anti-human IFN-γ antibody (11725-R209-B), anti-human IFN-γ antibody (11725-R238), HRP-conjugated anti-human IFN-γ antibody (11725-R209-H), recombinant human TNF-α protein (10602-HNAE), biotinylated anti-human TNF-α antibody (10602-MM08-B), and anti-human TNF-α antibody (10602-MM01) were obtained from SinoBiological.

Techniques:

Journal: Cell Reports Medicine

Article Title: CAR-T triggers TAM reeducation and adaptive anti-tumor response via TREM2 deficiency or CD40 agonist

doi: 10.1016/j.xcrm.2025.102539

Figure Lengend Snippet: The dual intervention of Trem2 -KO and GPC3-CAR-T induced profound remodeling of T cell ecosystems in both tumor and splenic microenvironments (A) UMAP clusters showed projection of cells onto a reference CD8 + T cell atlas (colored T cell subtypes) in tumor and spleen. (B) Proportion of the distinct CD8 + TILs and intrasplenic CD8 + T cell subtypes in tumor and spleen. (C) Violin plots revealed cell function based on each cell state scored by gene expressions of exhaustion, proliferation, cytotoxicity, and memory/naive phenotypes from CD8 + T subtypes in tumor, with medians and quartiles indicated. (D) Flow cytometry and quantification plots of IFN-γ + CD8 + T cells (effector CD8) in tumor and spleen. n = 8. (E) UMAP clusters showed projection of cells onto a reference CD4 + T cell atlas (colored T cell subtypes) in tumor and spleen. (F) Proportion of the distinct CD4 + TILs and intrasplenic CD4 + T cell subtypes in tumor and spleen. (G) Violin plots revealed cell function based on each cell state scored by gene expressions of exhaustion, proliferation, cytotoxicity, and memory/naive phenotypes from CD4 + T subtypes in tumor, with medians and quartiles indicated. (H) Flow cytometry and quantification plots of IFN-γ + CD4 + T cells in tumor and spleen. n = 8. (I and J) Multiple immunofluorescence images of tumor and spleen tissue between groups. Scale bars, 50 μm. Quantification of cells numbers from three random fields per mouse ( n = 8 mice/group). Data were represented by mean ± SEM. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001.

Article Snippet: Mouse IFN-γ (Interferon Gamma) ELISA Kit , MultiSciences , Cat#E-HSEL-M0007.

Techniques: Cell Function Assay, Flow Cytometry, Immunofluorescence

Journal: Cell Reports Medicine

Article Title: CAR-T triggers TAM reeducation and adaptive anti-tumor response via TREM2 deficiency or CD40 agonist

doi: 10.1016/j.xcrm.2025.102539

Figure Lengend Snippet: Trem2 -KO combined with IFN-γ promotes metabolic reprogramming in TAMs (A) Heatmap of KEGG pathways and differential gene set analysis of glycolysis, TCA, and fatty acid metabolism. Results were applied separately to Cxcl9 + and Spp1 + subtypes. (B) Heatmap of differential gene set analysis of glycolysis, TCA, and fatty acid metabolism. Results were applied separately to each group of CAR-T-treated WT or Trem2 -KO mice. (C) Schematic representation of in vitro tumor-educated macrophage model. BMDMs isolated from Trem2 -KO or WT mice were educated for 24 h in a transwell system with cancer cells, with or without IFN-γ (40 ng/mL) intervention. (D) Relative mRNA expression levels of Cxcl9 and Spp1 in BMDMs were quantified by RT-qPCR, and the ratio of Cxcl9 / Spp1 was calculated. (E) AMP and ATP concentrations in BMDMs were measured using dedicated assay kits, and the AMP/ATP ratio was calculated from these values. Relative mRNA expression levels of Ldha, Idh2, and Cpt1 in BMDMs were quantified by RT-qPCR. (F) Relative protein expression levels of indicated molecules were assessed by western blot (WB) and normalized to β-actin. (G, H, and O) BMDMs were pretreated for 2 h with the AMPK inhibitor compound C (20 μM; G), mTOR inhibitor rapamycin (1 μM; H), or SYK inhibitor R406 (1 μM; O), followed by 24 h co-culture with tumor cells in the presence or absence of IFN-γ (40 ng/mL). Protein expression was analyzed by WB and normalized to β-actin. (I and J) The real-time changes of OCR of BMDMs were stimulated with or without IFN-γ in the basal state and following the additions of oligomycin (Oligo), fluorocarbonyl cyanide phenylhydrazone (FCCP), etomoxir (Eto), and rotenone + antimycin A (Rot/AA). The average of basal OCR, maximal OCR, and Eto-sensitive OCR were revealed. (K and L) The real-time measurement of ECAR of BMDMs was stimulated with or without IFN-γ in the basal state and following the additions of glucose (Gluc), oligomycin (Oligo) and 2-deoxy-D-glucose (2-DG). The average of basal ECAR, maximal ECAR, and glycolytic reserve were revealed. (M and N) The relative protein expression levels of indicated molecular were assessed by WB and normalized to β-actin. (P) The iBMDMs were transiently transfected to overexpress SOCS1 for 48 h, followed by 24-h stimulation with or without IFN-γ (40 ng/mL). Indicated molecular protein expression levels were tested by WB and normalized to β-actin. Data were represented by mean ± SEM. ns, no significance, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001.

Article Snippet: Mouse IFN-γ (Interferon Gamma) ELISA Kit , MultiSciences , Cat#E-HSEL-M0007.

Techniques: In Vitro, Isolation, Expressing, Quantitative RT-PCR, Western Blot, Co-Culture Assay, Transfection

Journal: Cell Reports Medicine

Article Title: CAR-T triggers TAM reeducation and adaptive anti-tumor response via TREM2 deficiency or CD40 agonist

doi: 10.1016/j.xcrm.2025.102539

Figure Lengend Snippet: CD40 agonism triggers a metabolic reprogram of TAMs similar to that of Trem2 -KO (A) Heatmap of differential gene set analysis in TAM subpopulations. Genes were enriched in Mac_Cxcl9 cluster. (B) The network diagram illustrated the highly expressed genes in Mac_Cxcl9 cluster and their interactions with shared transcription factors. (C) Correlation analysis between the ratio of Cd40 + TAMs and Cxcl9 + TAMs . Each dot represented a single-cell sequencing sample of tumor and spleen. (D) BMDMs isolated from Trem2 -KO or WT mice were educated for 24 h in a transwell system with cancer cells, with or without IFN-γ intervention. Relative protein expression levels of CD40 were assessed by WB and normalized to β-actin. (E) Schematic representation of in vitro tumor-educated macrophage model. BMDMs isolated from WT mice were educated for 24 h in a transwell system with cancer cells, with or without IFN-γ (40 ng/mL) and CD40 agonism (crosslinked FGK45, 20 ng/mL) intervention. (F) Relative mRNA expression levels of Cxcl9 and Spp1 in BMDMs were quantified by RT-qPCR, and the ratio of Cxcl9/Spp1 was calculated. n = 3. (G and H) The real-time changes of OCR of BMDMs were stimulated with or without IFN-γ and CD40 agonism in the basal state and following the additions of oligomycin (Oligo), FCCP, etomoxir (Eto), and rotenone + antimycin A (Rot/AA). The average of basal OCR, maximal OCR, and Eto-sensitive OCR were revealed. n = 3. (I and J) The real-time measurement of ECAR of BMDMs were stimulated with or without IFN-γ and CD40 agonism in the basal state and following the additions of glucose (Gluc), oligomycin (Oligo), and 2-deoxy-D-glucose (2-DG). The average of basal ECAR, maximal ECAR, and glycolytic reserve were revealed. n = 3. (K, L, O, and P) BMDMs from WT mice were co-cultured with tumor cells in transwells for 24 h with or without IFN-γ (40 ng/mL) and CD40 agonism (crosslinked FGK45, 20 ng/mL) (K and O). For inhibitor studies, BMDMs were pre-treated with AMPK inhibitor (20 μM) (L) or STAT1 inhibitor (100 μM) (P) for 2 h prior to co-culture. Protein levels were analyzed by western blot and normalized to β-actin. (M and N) AMP and ATP concentrations in BMDMs were measured using dedicated assay kits, and the AMP/ATP ratio was calculated from these values. n = 3. (Q) Mechanism schema diagram. Data were represented by mean ± SEM. ns, no significance, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001.

Article Snippet: Mouse IFN-γ (Interferon Gamma) ELISA Kit , MultiSciences , Cat#E-HSEL-M0007.

Techniques: Sequencing, Isolation, Expressing, In Vitro, Quantitative RT-PCR, Cell Culture, Co-Culture Assay, Western Blot

Journal: Cell Reports Medicine

Article Title: CAR-T triggers TAM reeducation and adaptive anti-tumor response via TREM2 deficiency or CD40 agonist

doi: 10.1016/j.xcrm.2025.102539

Figure Lengend Snippet: The combination of CD40 agonism with GPC3-CAR-T therapy held promising application prospects in the treatment of HCC (A) Schematic procedure of HCC model constructed by Hepa1-6 cell orthotopic injection. GPC3-CART and CD40 agonist (FGK45) were injected. (B) Tumor representative photographs of different groups were shown. (C) Dot plots showed liver weight of mice in different groups. n = 8. (D) Survival curves of each group of mice. n = 8. (E and F) Multiple immunofluorescence images of TAMs, CD8 + , and CD4 + T cells in tumor tissue between groups. Scale bar, 50 μm. Quantification of cells numbers from three random fields per mouse ( n = 8 mice/group). Data were represented by mean ± SEM. ∗∗∗ p < 0.001. (G and H) Flow cytometry and quantification plots of IFN-γ + CD8 + T cells and IFN-γ + CD4 + T cells, as well as IFN-γ − CD8 + T cells and IFN-γ - CD4 + T cells from TILs in vivo . N = 8. Data were represented by mean ± SEM. ∗ p < 0.01, ∗∗∗ p < 0.001. (I) Schematic procedure of a therapeutic breast cancer liver metastasis model constructed by 4T1 cell expressing hTrop2 protein. Murine hTrop2-CAR-T and CD40 agonist (FGK45) were injected. (J) Tumor representative photographs of different groups were shown. (K) Dot plots showed liver weight of mice in different groups. n = 8. Data were represented by mean ± SEM. ns, no significance, ∗∗∗ p < 0.001. (L) Survival curves of each group of mice. n = 8. (M) Schematic representation of the transendothelial migration assay. TAMs were isolated from human HCC tissue. Sotigalimab (20 ng/mL) and h-IFN-γ (40 ng/mL) were added. Flow cytometry quantification of migrated CD8 + T cells (CellTrace Far Red labeled, n = 5). Data were represented by mean ± SEM. ns, no significance, ∗∗ p < 0.01, ∗∗∗ p < 0.001. (N) Schematic representation of the transendothelial migration assay. DTS was isolated from human GPC3 + HCC tissue. GPC3-CAR-T was added at indicated ratio, with or without sotigalimab. Flow cytometry quantification of migrated CD8 + T cells (CellTrace Far Red labeled, n = 5). Data were represented by mean ± SEM. ns, no significance, ∗∗∗ p < 0.001.

Article Snippet: Mouse IFN-γ (Interferon Gamma) ELISA Kit , MultiSciences , Cat#E-HSEL-M0007.

Techniques: Construct, Injection, Immunofluorescence, Flow Cytometry, In Vivo, Expressing, Migration, Isolation, Labeling