polyclonal anti tnfα  (R&D Systems)

Journal: Cell Reports Medicine

Article Title: LIFUS-driven engineered bacteria reprogram immunosuppressive niches via mechano-NOTCH signaling

doi: 10.1016/j.xcrm.2026.102658

Figure Lengend Snippet: Mechanical immunotherapy effect of LIFUS combined with VNP /ARG-GV for tumors (A) Schematic illustrating the five-cycle LIFUS + VNP /ARG-GVs treatment regimen over 21 days. (B and C) Repeated LIFUS activation of intratumoral VNP /ARG-GVs resulted in marked suppression of tumor growth compared with all control and monotherapy groups (∗∗∗∗ p < 0.0001; n = 6) (B). (D) Kaplan-Meier survival analysis demonstrating significantly prolonged survival in the combination group (median 47.5 days), with several mice surviving beyond 60 days. All control and monotherapy cohorts succumbed by days 30–40 (control and LIFUS alone: median 20 days, VNP /ARG: 27.5 days, VNP /ARG-GVs: 25 days, n = 10). (E and G) Flow cytometry analysis (E) showing differential immune remodeling across groups. The details of the flow cytometry gating strategy are shown in . LIFUS alone did not alter CD4 + /CD8 + infiltration. VNP /ARG monotherapy induced about 1.5-fold increase in CD8 + T cells. Combination treatment produced a pronounced expansion of cytotoxic CD8 + T cells (63.96% ± 4.90% vs. 14.31% ± 2.0% in control; ∗∗∗∗ p < 0.0001; n = 4) (G). (F and H) The combination group showed the strongest suppression of FOXP3 + Tregs (7.76% ± 1.60%), significantly lower than controls (28.38% ± 2.59%; ∗∗∗ p < 0.005; n = 4) (H). (I and J) ELISA quantification of effector cytokines. IFN-γ levels were highest in the combination group (37.4 ± 6.88 ng/mL), followed by LIFUS alone (12.1 ± 2.3 ng/mL) and control (5.6 ± 1.4 ng/mL) ( n = 4) (I). TNF-α expression followed a similar trend (1.37 ± 0.41 ng/mL vs. 0.42 ± 0.15 ng/mL and 0.18 ± 0.06 ng/mL) ( n = 4) (J). (K and L) Representative Ki67 and hematoxylin and eosin staining showing reduced proliferation and increased apoptosis across groups, with the strongest effects observed in the LIFUS + VNP /ARG-GVs cohort. Scale bars, 100 μm. Data are representative of three (B, D, I, and J) or two (G and H) independent experiments. Data are mean ± SD or chi-square (D). Statistical analysis was performed using one-way ANOVA (B and H–K), two-way ANOVA (G), or log rank test (D). See also .

Article Snippet: Anti-mouse TNF- α antibody , Proteintech , Cat# 17590-1-AP; RRID: AB_2271853.

Techniques: Activation Assay, Control, Flow Cytometry, Produced, Enzyme-linked Immunosorbent Assay, Expressing, Staining

Journal: Cell Reports Medicine

Article Title: LIFUS-driven engineered bacteria reprogram immunosuppressive niches via mechano-NOTCH signaling

doi: 10.1016/j.xcrm.2026.102658

Figure Lengend Snippet: Mechanobiology-enhanced adoptive T cell therapy improves control of primary and metastatic tumors (A) Schematic representation of the B16-OVA orthotopic melanoma model and the OT-1-based treatment strategy. The GVs (1 × 10 6 particles/mL) were injected intratumorally every cycle with LIFUS or without LIFUS before OT-1 cells (1 × 10 6 cells) were adoptively transferred later. (B) Tumor growth curves over 15 days. OT-1 +LIFUS treatment resulted in the most pronounced tumor inhibition, exhibiting significantly smaller tumor volumes than both OT-1 −LIFUS and control groups (266.7 ± 127.2 mm 3 vs. control 1,800 ± 1.003 mm 3 , ∗∗∗∗ p < 0.0001, vs. OT-1 −LIFUS 796.7 ± 613.2 mm 3 ; ∗∗∗ p < 0.005; n = 3). (C and D) Tumor (C) and body weight (D) measurements. The OT-1 +LIFUS group exhibited a marked reduction in tumor weight (0.18 ± 0.04 g vs. 0.71 ± 0.04 g in controls; ∗∗∗∗ p < 0.0001; n = 5). In contrast, body weight in OT-1 +LIFUS group remained stable throughout the 15-day treatment period (16.55 ± 0.15 g on day 15 vs. 16.57 ± 0.11 g at baseline; p > 0.05; n = 3). (E) Flow cytometric quantification of exhausted OT-1 cells within tumors. The OT-1 +LIFUS group exhibited a 94.1% reduction in PD-1 + Tim-3 + -co-expressing OT-1 cells compared with controls (2.55% ± 1.27% vs. 43.0% ± 4.60%; ∗∗∗∗ p < 0.0001; n = 5). (F) Intracellular cytokine staining of TNF-α and IFN-γ. OT-1 +LIFUS significantly enhanced effector cytokine production relative to both comparison groups (∗∗∗∗ p < 0.0001; n = 5). (G) Proliferative activity of intratumoral OT-1 cells, assessed by Ki-67 expression. OT-1 +LIFUS showed the highest proportion of Ki-67 + OT-1 cells (63.20% ± 8.43%), indicating improved expansion and functional fitness (∗∗∗∗ p < 0.0001; n = 5). (H) Schematic of the 4T1-luciferase lung metastasis model and CAR-T-based treatment strategy. CAR-T cells were incubated with GVs (1 × 10 6 particles/mL, 1:1 ratio) for 10 min and exposed to LIFUS 60 s before i.v. injection. (I) Bioluminescence analysis of metastatic tumor burden. Photon flux in the CAR-T +LIFUS group was reduced the most relative to controls (mean (4.23 ± 2.09) × 10 7 ps −1 cm −2 sr −1 at the endpoint), consistent with substantially attenuated metastatic progression ( n = 5). (J) Representative longitudinal bioluminescence imaging. Detectable thoracic luciferase signal at day 0 confirmed successful establishment of lung metastases. Metastatic burden increased rapidly in controls beginning at day 5, whereas CAR-T +LIFUS exhibited minimal signal on days 10–15 and near-complete regression by day 25 ((1.6 ± 9.3) × 10 4 ps −1 cm −2 sr −1 vs. (5.3 ± 2.3) × 10 7 ps −1 cm −2 sr −1 in the control group, ∗∗∗∗ p < 0.0001). Tails were covered to prevent signal interference from freshly administered D-luciferin ( n = 5) (see D). (K) Kaplan-Meier survival analysis. CAR-T +LIFUS markedly prolonged survival (median 63.5 days) compared with CAR-T monotherapy (47.5 days; ∗∗ p < 0.01) and controls (35.5 days; ∗∗∗ p < 0.005) ( n = 6). Data are representative of two (B–G, I, and K) independent experiments. Data are mean ± SD (B–G), the mean value (I), or the Chi square (K). Statistical analysis was performed using one-way ANOVA (C and E–G), two-way ANOVA (B and D), or log rank test (K). Also see D–S5F.

Article Snippet: Anti-mouse TNF- α antibody , Proteintech , Cat# 17590-1-AP; RRID: AB_2271853.

Techniques: Control, Injection, Inhibition, Expressing, Staining, Comparison, Activity Assay, Functional Assay, Luciferase, Incubation, Imaging

Journal: Journal of Pain Research

Article Title: Unexpected Role of TNFα Signaling in the Resolution of Postoperative Pain in Mice

doi: 10.2147/JPR.S543971

Figure Lengend Snippet: Neutralization of TNFα prolonged pain hypersensitivity induced by surgical incision. Mechanical sensitivity was assessed with von Frey filaments. ( A ) Neutralizing polyclonal anti-TNFα (5 µg) was injected at 2 and 24 h after incision (Control n= 4M and 3F, anti-TNFα n= 2M and 4F). Two-way ANOVA with Sidak’s correction time x treatment interaction: F (5, 55) = 5.0, p=0.0008. ( B ) Neutralizing monoclonal anti-TNFα (5 µg) was injected at 2 and 24 h after incision (Control n= 3M and 3F, anti-TNFα n= 3M and 3F). Two-way ANOVA with Sidak’s correction time x treatment interaction: F(2.6,25.9)= 18.6, p<0.0001). On the graph * = p<0.05, and ** = p<0.01. BL: indicates baseline measures performed before the surgery.

Article Snippet: Intraplantar administration of neutralizing polyclonal anti-TNFα (RD System, #AF-410-NA, 5μg), isotype control IgG (Cell signaling Technology #7076S), monoclonal anti-TNFα clone XT3 (BioXCell, #BE0058, 5μg), isotype control InVivoPlus mouse IgG1 clone MOPC-21 (BioXCell, #BP0083), Etanercept (MedChemExpress #HY-108847) and PBS.

Techniques: Neutralization, Injection, Control

Journal: Journal of Pain Research

Article Title: Unexpected Role of TNFα Signaling in the Resolution of Postoperative Pain in Mice

doi: 10.2147/JPR.S543971

Figure Lengend Snippet: Absence of sex difference on the impact of TNFα signaling in acute postoperative pain resolution. ( A ) Effect of polyclonal antibody on mechanical pain corresponding to . ( B ) Effect of monoclonal antibody on mechanical pain corresponding to . ( C ) Effect of etanercept on mechanical pain corresponding to . ( D ) Effect of etanercept on thermal pain corresponding to . Effect of sex assessed by two-way ANOVA with multiple comparison test correction ( A ) p=0.14, ( B ) p=0.22, ( C ) p=0.59, and ( D ) p=0.53. BL: indicates baseline measures performed before the surgery.

Article Snippet: Intraplantar administration of neutralizing polyclonal anti-TNFα (RD System, #AF-410-NA, 5μg), isotype control IgG (Cell signaling Technology #7076S), monoclonal anti-TNFα clone XT3 (BioXCell, #BE0058, 5μg), isotype control InVivoPlus mouse IgG1 clone MOPC-21 (BioXCell, #BP0083), Etanercept (MedChemExpress #HY-108847) and PBS.

Techniques: Comparison

Journal: Biological Research

Article Title: Female prediabetic rats are protected from vascular dysfunction: the role of nitroso and sulfide signaling

doi: 10.1186/s40659-024-00575-1

Figure Lengend Snippet: Gene and protein expressions of inflammatory factors in female Wistar and HTG rats

Article Snippet: Afterward, the membranes were incubated with a rabbit polyclonal anti-eNOS antibody (Abcam, Cambridge, UK; dilution 1:1000), a rabbit polyclonal anti-iNOS antibody (Proteintech ® , Manchester, UK; dilution 1:1000), a rabbit polyclonal anti-CBS antibody (Proteintech ® , Manchester, UK; dilution 1:3000), a mouse monoclonal anti-CSE antibody (Proteintech ® , Manchester, UK; dilution 1:5000), a mouse monoclonal anti-TNFα antibody (Proteintech ® , Manchester, UK; dilution 1:3000), a mouse monoclonal anti-NFκB antibody (Cell Signaling, Danvers, MA, USA; dilution 1:1000), a rabbit polyclonal anti-Mcp-1 antibody (Abcam, Cambridge, UK; dilution 1:1000), a rabbit polyclonal anti-ICAM antibody (Proteintech ® , Manchester, UK; dilution 1:1000), or a rabbit polyclonal anti-COX2 antibody (Proteintech ® , Manchester, UK; dilution 1:500) overnight at 4 °C.

Techniques:

Journal: BMC Biotechnology

Article Title: Anti-inflammatory potential of aspergillus unguis SP51-EGY: TLR4-dependent effects & chemical diversity via Q-TOF LC-HRMS

doi: 10.1186/s12896-024-00890-1

Figure Lengend Snippet: mRNA sequences used for qPCR

Article Snippet: Cyclooxygenase-2 (COX-2) Polyclonal Antibody, Mouse TNF-α, and Mouse IL-6 ELISA Kits were purchased from Elabscience (Wuhan, China).

Techniques: Sequencing

Journal: BMC Biotechnology

Article Title: Anti-inflammatory potential of aspergillus unguis SP51-EGY: TLR4-dependent effects & chemical diversity via Q-TOF LC-HRMS

doi: 10.1186/s12896-024-00890-1

Figure Lengend Snippet: Expression levels of iNOS, COX-2, TNF-α, and IL-6 mRNA in LPS-stimulated RAW 264.7 macrophages. Fungal extracts in medium A-D (10 µg/mL) were used for treatment of LPS-stimulated RAW 264.7 macrophages (LPS concentration: 10 ng/mL). The mRNA levels of iNOS, COX-2, TNF-α and IL-6 were measured using qPCR by the comparative method (2 −ΔΔCT ). Results are presented as the mean ± SE ( n = 3). Statistical significance was calculated by one-way ANOVA followed by Student–Newman–Keuls post-hoc test. $ P < 0.05 vs. control cells. * P < 0.05 vs. LPS-stimulated cells

Article Snippet: Cyclooxygenase-2 (COX-2) Polyclonal Antibody, Mouse TNF-α, and Mouse IL-6 ELISA Kits were purchased from Elabscience (Wuhan, China).

Techniques: Expressing, Concentration Assay, Control

Journal: BMC Biotechnology

Article Title: Anti-inflammatory potential of aspergillus unguis SP51-EGY: TLR4-dependent effects & chemical diversity via Q-TOF LC-HRMS

doi: 10.1186/s12896-024-00890-1

Figure Lengend Snippet: Determination of protein levels of the pro-inflammatory cytokines TNF-α and IL-6 in the cell culture medium of the LPS-stimulated RAW 264.7 macrophages using ELISA. The selected fungal extracts (Sh F and St Cell in medium B and Sh Cell in medium C) showed a significant reduction of the LPS-stimulated upregulation of the pro-inflammatory cytokines TNF-α ( A ) and IL-6 ( B ). Results are presented as the mean ± SE ( n = 3). Statistical significance was calculated by one-way ANOVA followed by Student–Newman–Keuls post-hoc test. $ P < 0.05 vs. control cells. * P < 0.05 vs. LPS-stimulated cells

Article Snippet: Cyclooxygenase-2 (COX-2) Polyclonal Antibody, Mouse TNF-α, and Mouse IL-6 ELISA Kits were purchased from Elabscience (Wuhan, China).

Techniques: Cell Culture, Enzyme-linked Immunosorbent Assay, Control