il 1β  (R&D Systems)

Journal: Journal of Inflammation Research

Article Title: The Relationship Between Decreased Serum Klotho Protein and Cognitive Impairment in Patients After Abdominal Surgery

doi: 10.2147/JIR.S593405

Figure Lengend Snippet: The correlation between serum Klotho and clinical indicators. Pearson correlation test was performed between Klotho with ( A ) duration of anesthesia, ( B ) postoperative MoCA scores, ( C ) postoperative CRP, ( D ) postoperative IL-1β, and ( E ) postoperative IL-6 in all 93 cases of patients after abdominal surgery.

Article Snippet: CRP (catalog no. SEKH-0138) and Klotho (catalog no. SEKH-0280) ELISA kits were purchased from Solarbio (Beijing, China), while IL-1β (catalog no. DLB50) and IL-6 (catalog no. D6050B) ELISA kits were obtained from R&D Systems (Minneapolis).

Techniques:

Journal: Journal of Inflammation Research

Article Title: The Relationship Between Decreased Serum Klotho Protein and Cognitive Impairment in Patients After Abdominal Surgery

doi: 10.2147/JIR.S593405

Figure Lengend Snippet: Proposed mechanism linking decreased postoperative serum Klotho levels to postoperative cognitive dysfunction (POCD) after abdominal surgery. Surgery and prolonged anesthesia lead to a reduction in serum Klotho levels, which contributes to increased oxidative stress and systemic inflammation, as indicated by elevated levels of CRP, IL-1β, and IL-6. These processes promote neuroinflammation and synaptic dysfunction, resulting in cognitive decline and POCD. Arrows indicate the direction of the effects, illustrating how decreased Klotho mediates the relationship between perioperative stress, inflammation, and postoperative cognitive impairment.

Article Snippet: CRP (catalog no. SEKH-0138) and Klotho (catalog no. SEKH-0280) ELISA kits were purchased from Solarbio (Beijing, China), while IL-1β (catalog no. DLB50) and IL-6 (catalog no. D6050B) ELISA kits were obtained from R&D Systems (Minneapolis).

Techniques:

Journal: Frontiers in Immunology

Article Title: Secretory VSMC-derived THBS1 promotes macrophage M1 polarization and inflammation in intracranial aneurysms via the CD47/NF-κB axis

doi: 10.3389/fimmu.2026.1790488

Figure Lengend Snippet: Secretory VSMCs promote macrophage M1 polarization and inflammatory activation through the THBS1/CD47/NF-κB axis. (A) ELISA quantification of THBS1 secretion from HA-VSMCs before and after TGF-β1/PDGF-BB stimulation and following shTHBS1 transduction. (B–D) qRT-PCR analysis of macrophage IL-1β, TNF-α, and iNOS mRNA in the Transwell co-culture system. (E, F) ELISA quantification of IL-1β and TNF-α secretion in co-culture supernatants. (G, H) Flow cytometric analysis of CD86 + macrophages and NF-κB transcriptional activity (Dual-luciferase reporter assay). All in vitro experiments were independently performed in three biological replicates (n = 3). Data are presented as mean ± standard deviation (SD). Statistical comparisons among multiple groups were performed using one-way analysis of variance (ANOVA) followed by Tukey’s post hoc multiple-comparison test. *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: At the protein secretion level, commercially available Quantikine ® ELISA kits (R&D Systems, DY201 for IL-1β; DY210 for TNF-α) were used to quantify IL-1β and TNF-α concentrations in the lower-chamber supernatants following the manufacturer’s instructions.

Techniques: Activation Assay, Enzyme-linked Immunosorbent Assay, Transduction, Quantitative RT-PCR, Co-Culture Assay, Activity Assay, Luciferase, Reporter Assay, In Vitro, Standard Deviation, Comparison

Journal: Science Advances

Article Title: Inflammatory arthritis irAE may represent a unique autoimmune disease primarily driven by T cells but likely not autoantibodies

doi: 10.1126/sciadv.aea4262

Figure Lengend Snippet: ( A to C ) Human naïve B cells were isolated from PBMCs and cultured under indicated conditions for 7 days; pembrolizumab or isotype control IgG4 (IgG4 Iso) was added on day 2; n = 8. (A) Expression of CD38 and CD27 on B cells. Right: Percentages of CD27 + CD38 − and CD27 + CD38 + B cells. (B) Expression of CD138 on CD27 + CD38 + B cells. Right: Percentage of CD138 + CD27 + CD38 + B cells. (C) Different immunoglobulin isotypes were measured in the culture supernatant of (A) and (B) by multiplex assay. ( D to F ) B cells from Humanized PD-1 (HuPD-1) mice were isolated and cultured with LPS, IL-4, BAFF, or ODN 2006, anti-IgM, IL-21, IL-4 or R848, anti-IgM, anti-CD40, IL-21, and IFN-γ for 3 days; pembrolizumab or isotype control was added on day 1. (D) Expression of IgG2c on activated B cells. FSC-H, Forward scatter height. Right: Percentage of IgG2c + B cells; n = 5. (E) Expression of IgG1 on activated B cells. Right: Percentage of IgG1 + B cells; n = 5. (F) Different immunoglobulin isotypes in the supernatant were measured by multiplex assay; n = 5. Data in graphs represent mean ± SEM. Significance was tested by two-way ANOVA.

Article Snippet: For CXCL13, IL-21, and CX3CL1 measurements, the following kits were used: Human CXCL13/BLC/BCA-1 Quantikine Enzyme-Linked Immunosorbent Assay (ELISA) (R&D Systems, catalog no. DCX130), Human IL-21 DuoSet ELISA (R&D Systems, catalog no. DY8879-05), and Human CX3CL1/Fractalkine DuoSet ELISA (R&D Systems, DY365); all steps were performed according to the manufacturer’s instructions.

Techniques: Isolation, Cell Culture, Control, Expressing, Multiplex Assay

Journal: Science Advances

Article Title: Inflammatory arthritis irAE may represent a unique autoimmune disease primarily driven by T cells but likely not autoantibodies

doi: 10.1126/sciadv.aea4262

Figure Lengend Snippet: ( A to G ) CD8 + T cells isolated from healthy donor were cultured in the plate coated with anti–human CD3/CD28 (10 μg/ml) for indicated days in the presence of vehicle control (control), IFN-α (100 ng/ml), IL-6 (100 ng/ml), IL-12 (100 ng/ml), or the combination. (A) Summaries of the percentage of CD38 + CD127 − CD8 + T cells at days 1 and 5. (B) MFI of CD69 (day 5). (C) MFI of CD25 (day 5). (D) Activated CD8 + T cells were restimulated with PMA, ionomycin, and monensin for 5 hours; expression of granzyme B and IFN-γ in CD8 + T cells was examined. [(E) to (G)] CD8 + T cells were cultured for 5 days. MFIs (all relative to those under control condition) of MTDR (E), MTG (F), and GluCy5 (G) were summarized. ( H to L ) CD4 + T cells were isolated from HC, cultured in the plate coated with anti–human CD3/CD28 (10 μg/ml) for indicated days in the presence of IFN-α (100 ng/ml), IL-6 (100 ng/ml), IL-12 (100 ng/ml), control, or the combination of them. (H) Activated CD4 + T cells were restimulated with PMA, ionomycin, and monensin for 5 hours, and IL-21 level was examined in the CD45RA − CD4 + T cells. (I) Activated CD4 + T cells were restimulated with plate-coated anti–human CD3/CD28 (10 μg/ml) with monensin for 6 hours, and expression of CXCL13 was measured in CD45RA − CD4 + T cells. (J) CD38 and CXCR5 were examined on CD4 + T cells at day 5. (K) Percentage of CXCL13 + cells in indicated T cells at day 5. (L) Percentage of IL-21 + cells in indicated T cells at day 5. Data in graphs represent mean ± SEM. Significance was tested by one-way ANOVA.

Article Snippet: For CXCL13, IL-21, and CX3CL1 measurements, the following kits were used: Human CXCL13/BLC/BCA-1 Quantikine Enzyme-Linked Immunosorbent Assay (ELISA) (R&D Systems, catalog no. DCX130), Human IL-21 DuoSet ELISA (R&D Systems, catalog no. DY8879-05), and Human CX3CL1/Fractalkine DuoSet ELISA (R&D Systems, DY365); all steps were performed according to the manufacturer’s instructions.

Techniques: Isolation, Cell Culture, Control, Expressing

Journal: Science Advances

Article Title: Inflammatory arthritis irAE may represent a unique autoimmune disease primarily driven by T cells but likely not autoantibodies

doi: 10.1126/sciadv.aea4262

Figure Lengend Snippet: ( A to G ) PBMCs from the patients with irAE were cultured in the plate coated with anti-CD3 and anti-CD28 (10 μg/ml) in the presence of IgG1 isotype control or anti–human IL-6R (50 μg/ml), anti–human IL-12p40 (50 μg/ml), and anti–human IFNAR1 (50 μg/ml) for 3 days; n = 9. (A) Expression of CD38 and CD127 on CD8 + T cells. Right: Percentage of CD38 + CD127 − CD8 + T cells. (B and C) CD8 + T cells activated for 5 days were restimulated with PMA, ionomycin, and monensin for 5 hours. (B) Expression of granzyme B and IFN-γ. Right: Percentage of granzyme B + IFN-γ + CD8 + T cells. (C) Expression of perforin and IFN-γ. Right: Percentage of perforin + IFN-γ + CD8 + T cells. [(D) and (E)] MFIs of GluCy5 (D), TMRM, and MTDR (E) in CD8 + T cells were presented. [(F) and (G)] CD4 + T cells activated for 5 days were restimulated with PMA, ionomycin, and monensin for 5 hours. (F) Expression of IL-21 and IL-2. Right: Percentage of IL-21 + IL-2 + CD4 + T cells. (G) Expression of CD38 and CXCR5 on CD4 + T cells. Right: Percentage of CD38 + CXCR5 − CD4 + T cells. Data in graphs represent mean ± SEM. Significance was tested paired Student’s t test [(A) to (G)].

Article Snippet: For CXCL13, IL-21, and CX3CL1 measurements, the following kits were used: Human CXCL13/BLC/BCA-1 Quantikine Enzyme-Linked Immunosorbent Assay (ELISA) (R&D Systems, catalog no. DCX130), Human IL-21 DuoSet ELISA (R&D Systems, catalog no. DY8879-05), and Human CX3CL1/Fractalkine DuoSet ELISA (R&D Systems, DY365); all steps were performed according to the manufacturer’s instructions.

Techniques: Cell Culture, Control, Expressing

Journal: Central-European Journal of Immunology

Article Title: p65 enhanced METTL3-mediated m 6 A methylation of HMGB1 to promote microglia M1 polarization in sepsis-associated encephalopathy

doi: 10.5114/ceji.2025.154882

Figure Lengend Snippet: p65, METTL3, and HMGB1 increased in lipopolysaccharide (LPS)-induced HMC3 cells. HMC3 cells were stimulated with LPS. A ) CCK-8 kit was assessed using cell viability. B ) The proportion of CD86+CD11b+ cells in HMC3 cells was evaluated using flow cytometry. Data are presented as means ± SD. Significance levels were: * p < 0.05, ** p < 0.01, and *** p < 0.001 C ) Inflammatory cytokines including TNF-α, IL-1β, and IL-6 were measured by ELISA kits. D ) Western blot was performed to detect the levels of iNOS, p-p65, METTL3, and HMGB1. All experiments were performed in triplicate. Data are presented as means ± SD. Significance levels were: * p < 0.05, ** p < 0.01, and *** p < 0.001

Article Snippet: Concentrations of inflammatory cytokines, such as tumor necrosis factor α (TNF-α, #DTA00D), interleukin 1β (IL-1β, #DLB50), and IL-6 (#D6050), were assessed using ELISA kits from R&D Systems located in Minneapolis, MN, USA.

Techniques: CCK-8 Assay, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Western Blot

Journal: Central-European Journal of Immunology

Article Title: p65 enhanced METTL3-mediated m 6 A methylation of HMGB1 to promote microglia M1 polarization in sepsis-associated encephalopathy

doi: 10.5114/ceji.2025.154882

Figure Lengend Snippet: p65 activated METTL3 promoted LPS-induced microglia M1 polarization HMC3 cells were transfected with sh-METTL3 and/or oe-p65 and stimulated with LPS. A ) qRT-PCR was used to detect the expression of p65 and METTL3. B ) A CCK-8 kit was used to assess cell viability. C ) Flow cytometry was used to evaluate the proportion of CD86+CD11b+ cells among HMC3 cells. Data are presented as means ± SD. Significance levels were considered as follows: * p < 0.05, ** p < 0.01, and *** p < 0.001 C ) Flow cytometry was used to evaluate the proportion of CD86+CD11b+ cells among HMC3 cells. Data are presented as means ± SD. Significance levels were considered as follows: * p < 0.05, ** p < 0.01, and *** p < 0.001 D ) TNF-α, IL-1β, and IL-6 levels were measured by ELISA kits. E ) Western blot was performed to detect iNOS, p-p65, and METTL3 expression. All experiments were performed in triplicate. Data are presented as means ± SD. Significance levels were considered as follows: * p < 0.05, ** p < 0.01, and *** p < 0.001

Article Snippet: Concentrations of inflammatory cytokines, such as tumor necrosis factor α (TNF-α, #DTA00D), interleukin 1β (IL-1β, #DLB50), and IL-6 (#D6050), were assessed using ELISA kits from R&D Systems located in Minneapolis, MN, USA.

Techniques: Transfection, Quantitative RT-PCR, Expressing, CCK-8 Assay, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Western Blot

Journal: Central-European Journal of Immunology

Article Title: p65 enhanced METTL3-mediated m 6 A methylation of HMGB1 to promote microglia M1 polarization in sepsis-associated encephalopathy

doi: 10.5114/ceji.2025.154882

Figure Lengend Snippet: METTL3 up-regulated HMGB1 and enhanced LPS-induced microglia M1 polarization. HMC3 cells were transfected with sh-HMGB1 and/or oe-METTL3 and stimulated with LPS. A ) qRT-PCR was used to detect the expression of METTL3 and HMGB1. B ) A CCK-8 kit was used to assess cell viability. C ) Flow cytometry was used to evaluate the proportion of CD86+CD11b+ cells among HMC3 cells. Data are presented as means ± SD. Significance levels were considered as follows: * p < 0.05, ** p < 0.01, and *** p < 0.001 C ) Flow cytometry was used to evaluate the proportion of CD86+CD11b+ cells among HMC3 cells. Data are presented as means ± SD. Significance levels were considered as follows: * p < 0.05, ** p < 0.01, and *** p < 0.001 D ) TNF-α, IL-1β, and IL-6 levels were measured by ELISA kits. E ) Western blot was performed to detect iNOS, HMGB1, and METTL3 expression. All experiments were performed in triplicate. Data are presented as means ± SD. Significance levels were considered as follows: * p < 0.05, ** p < 0.01, and *** p < 0.001

Article Snippet: Concentrations of inflammatory cytokines, such as tumor necrosis factor α (TNF-α, #DTA00D), interleukin 1β (IL-1β, #DLB50), and IL-6 (#D6050), were assessed using ELISA kits from R&D Systems located in Minneapolis, MN, USA.

Techniques: Transfection, Quantitative RT-PCR, Expressing, CCK-8 Assay, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Western Blot