Journal: Brain, behavior, and immunity

Article Title: STING recognition of viral dsDNA by nociceptors mediates pain in mice.

doi: 10.1016/j.bbi.2024.07.013

Figure Lengend Snippet: Fig. 4. Virus-derived dsDNA-induced pain behaviors rely on TRPV1 expression. (A) Illustration depicting the canonical STING signaling and the hypothesized dependency on TRP channels. (B) Nocifensive behaviors and mechanical sensitivity induced by intraplantar (i.pl.) injection of HSV-60 (5 µg/20 µl) after intra- peritoneal (i.p.) injection of STING agonists or a vehicle control (n = 6 male mice/group). (C-E) Nocifensive behaviors i.pl. injection of HSV-60 (5 µg/20 µl) in mice treated with TBK1 antagonist GSK8612 (5 mg/kg, i.p., C), IFNß antibody (10 µg, i.pl.), TRPA1 inhibitor HC-030031 (35 mg/kg, i.p., E) or TRPM8 inhibitor AMG2850 (30 mg/kg, i.p., E), compared to their respective controls (n = 4–5 male mice/group). (F) Nocifensive behaviors and mechanical sensitivity induced by i.pl. injection of HSV-60 (5 µg/20 µl) after i.p. injection of TRPV1 inhibitor AMG9810 (100 mg/100 ml) or vehicle control (n = 5 male mice/group). Statistical analysis: two-tailed unpaired Student’s t-test (C-E, F; nocifensive behaviors), one-way ANOVA followed by Dunnett’s multiple comparisons test (B; nocifensive behaviors), two-way ANOVA followed by Dunnett’s multiple comparisons test (B; mechanical sensitivity) or ˇSíd´ak’s multiple comparisons test (F; mechanical sensitivity). Error bars indicate mean ± s.e.m. and *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

Article Snippet: An ELISA kit for IFNß (# 424101) and IFNα (#421151) were purchased from PBL Assay Science.

Techniques: Virus, Derivative Assay, Expressing, Injection, Control, Two Tailed Test

Journal: Translational Oncology

Article Title: IL1RA inhibits the progression of oral squamous cell carcinoma by mediating type Ⅰ interferon response

doi: 10.1016/j.tranon.2025.102428

Figure Lengend Snippet: IL1RA is associated with oncogenic signaling pathways and type I interferon response in OSCC in vitro . A. Volcano plots visualizing 132 DEGs between CAL27 cells transfected with NC and oeIL1RA. Down-regulated, up-regulated and non-regulated genes were labeled in blue, red, and grey colors, respectively. B. GSEA showed less enriched cancer-related features or processes in CAL27 cells overexpressing IL1RA. C. GSEA showed that IL1RA overexpression was significantly associated with the type I interferon response. D. A Venn diagram visualizing 45 (61.6 %) OSCC patients simultaneously carrying mutations in the IL1RA, IFNA , and IFNB genes in the TCGA dataset.

Article Snippet: Slices were then incubated with diluted primary antibodies against IL1RA (NBP1-32568, Novusbio, 1:600), E-Cadherin (#13116, CST, 1:400), N-Cadherin (#3195, CST, 1:100), Vimentin (#5741, CST, 1:100), IFNA (18013-AP, Proteintech, 1:400), and IFNB (27506-AP, Proteintech, 1:400) overnight at 4° C, and the secondary antibody (horseradish peroxidase-labeled goat anti-rabbit) at room temperature for 2 h. Cell nuclei were dyed with a DAB staining solution.

Techniques: Protein-Protein interactions, In Vitro, Transfection, Labeling, Over Expression

Journal: Translational Oncology

Article Title: IL1RA inhibits the progression of oral squamous cell carcinoma by mediating type Ⅰ interferon response

doi: 10.1016/j.tranon.2025.102428

Figure Lengend Snippet: Overexpression of IL1RA up-regulates type I interferon proteins in OSCC in vitro . A. H&E (the first lane) and IHC staining (the latter three lanes) of IFNA and IFNB in OSCC specimens of the high and low IL1RA expression groups (n=30, scale bar=100 μm). B-C. The mRNA (B) and protein expressions (C) of IFNA and IFNB in OSCC specimens of the high and low IL1RA expression groups. * P <0.05, and ** P <0.01 vs. NC group.

Article Snippet: Slices were then incubated with diluted primary antibodies against IL1RA (NBP1-32568, Novusbio, 1:600), E-Cadherin (#13116, CST, 1:400), N-Cadherin (#3195, CST, 1:100), Vimentin (#5741, CST, 1:100), IFNA (18013-AP, Proteintech, 1:400), and IFNB (27506-AP, Proteintech, 1:400) overnight at 4° C, and the secondary antibody (horseradish peroxidase-labeled goat anti-rabbit) at room temperature for 2 h. Cell nuclei were dyed with a DAB staining solution.

Techniques: Over Expression, In Vitro, Immunohistochemistry, Expressing

Journal: Translational Oncology

Article Title: IL1RA inhibits the progression of oral squamous cell carcinoma by mediating type Ⅰ interferon response

doi: 10.1016/j.tranon.2025.102428

Figure Lengend Snippet: IL1RA promotes the expressions of IFNA and IFNB in the OSCC xenografts and their release in OSCC cells. A. IHC staining of positive expressions of IFNA and IFNB in OSCC xenografts of oeIL1RA group and NC group in vivo (scale bar=100 μm). B. The mRNA levels of IFNA and IFNB in OSCC xenografts of the oeIL1RA group and NC group in vivo . C. ELISA showed contents of IFNA and IFNB in the cell supernatant of the oeIL1RA group and NC group in vitro . * P <0.05, and ** P <0.01 vs. NC group.

Article Snippet: Slices were then incubated with diluted primary antibodies against IL1RA (NBP1-32568, Novusbio, 1:600), E-Cadherin (#13116, CST, 1:400), N-Cadherin (#3195, CST, 1:100), Vimentin (#5741, CST, 1:100), IFNA (18013-AP, Proteintech, 1:400), and IFNB (27506-AP, Proteintech, 1:400) overnight at 4° C, and the secondary antibody (horseradish peroxidase-labeled goat anti-rabbit) at room temperature for 2 h. Cell nuclei were dyed with a DAB staining solution.

Techniques: Immunohistochemistry, In Vivo, Enzyme-linked Immunosorbent Assay, In Vitro

Journal: PLoS ONE

Article Title: Role of Lactobacillus pentosus Strain b240 and the Toll-Like Receptor 2 Axis in Peyer's Patch Dendritic Cell-Mediated Immunoglobulin A Enhancement

doi: 10.1371/journal.pone.0091857

Figure Lengend Snippet: (A) PP cells (1.5×10 6 cells) were cultured with saline (open circles), 1.2×10 6 counts of heat-killed b240 (closed squares), or 1.2 × 10 7 counts of heat-killed b240 (closed circles) for 1, 3, 5, and 7 days. (B, C) In the presence or absence of heat-killed b240 (4.7×10 6 counts), PP cells (5.8×10 5 cells) were cultured with (B) anti-IL-6 mAb (10 μg/ml), anti-IFN-γ mAb (10 μg/ml), anti-TNF mAb (10 μg/ml), rat IgG1 k isotype control (10 μg/ml), (C) LE540 (1 μM), BCMA-Ig+ TACI-Ig (5 μg/ml each), dimethyl sulfoxide, or human IgG1 Fc antibody (10 μg/ml) for 4 days. The stimulation index of each sample was calculated (for example, (b240-treatment and anti-IL-6 Ab treatment)/(saline-treatment and anti-IL-6 Ab treatment) is the stimulation index for anti-IL-6 Ab treatment). (D) PP cells (5.8×10 5 cells) were cultured with a low dose (light gray), medium dose (dark gray), and high dose (black) of rIL-6 (0.4, 2, or 10 ng/ml), rIFN-γ (0.6, 3, or 15 ng/ml), rTNF (0.08, 0.4, or 2 ng/ml), or heat-killed b240 (4.7×10 6 counts) for 4 days. IgA or cytokine in the culture supernatants was determined by ELISA or CBA. Data are expressed as mean ± SEM (n = 3). (A, B) * P <0.05 versus control group by Dunnett's test. (C) Student's t -test was conducted. (D) Statistical analysis was not conducted. Data are representative of 2 independent experiments producing similar results.

Article Snippet: IFN-α and IFN-β were measured using Mouse Interferon Alpha and Mouse Interferon Beta ELISA kit (PBL Biomedical Laboratories, Piscataway, NJ, USA), respectively.

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

Journal: PLoS ONE

Article Title: Role of Lactobacillus pentosus Strain b240 and the Toll-Like Receptor 2 Axis in Peyer's Patch Dendritic Cell-Mediated Immunoglobulin A Enhancement

doi: 10.1371/journal.pone.0091857

Figure Lengend Snippet: (A, C) In the presence (black) or absence (white) of heat-killed b240 (1.6×10 6 counts), (A) purified WT PP IgD + cells (2×10 5 cells/well) were cultured with or without purified WT PP CD11c + B220 − DCs (1×10 5 cells/well), (B) purified WT PP CD11c + B220 − DCs and purified WT PP IgD + B cells were co-cultured with (black) or without (white) anti-IL-6 mAb (10 μg/ml) in the presence or absence of heat-killed b240 (1.6×10 6 counts), and (C) purified WT or TLR2 −/− PP CD11c + B220 − DCs and purified WT or TLR2 −/− PP IgD + B cells were co-cultured for 7 days. IgA and IL-6 in the culture supernatants were determined by ELISA and CBA, respectively. Data are expressed as mean ± SEM (n = 1–3). ND, not detected. * P <0.05 versus (A) saline group by Welch's t -test, (B) isotype Ig group by Student's t- t est, (C) WT DCs + WT B cells + b240 co-culture by Dunnett's test. Data are representative of 2 independent experiments producing similar results.

Article Snippet: IFN-α and IFN-β were measured using Mouse Interferon Alpha and Mouse Interferon Beta ELISA kit (PBL Biomedical Laboratories, Piscataway, NJ, USA), respectively.

Techniques: Purification, Cell Culture, Enzyme-linked Immunosorbent Assay, Saline, Co-Culture Assay

Journal: Biochemical Journal

Article Title: Suppression of interferon β gene transcription by inhibitors of bromodomain and extra-terminal (BET) family members

doi: 10.1042/BJ20141523

Figure Lengend Snippet: RAW cells were incubated for 1 h without (−) or with (+) 1.0 μM JQ1 or 1.0 μM BI2536 and then stimulated for 8 h without (−) or with (+) poly(I:C) (10 μg/ml) ( A and C ) or for 4 h without (−) or with (+) LPS (100 ng/ml) ( B and D ). At each time point, the total RNA was extracted from the cells and Ifnb mRNA was quantified by qPCR ( A and B ) and the concentration of IFNβ in the cell culture medium was determined by ELISA ( C and D ). Results are means+S.E.M for triplicate determinations. ( A ) and ( B ) show the fold increase in mRNA levels relative to the values measured in cells that had not been stimulated with LPS or poly(I:C).

Article Snippet: The level of secreted IFNβ in the cell culture medium was determined using the Verikine mouse and human IFNβ ELISA kits (PBL Interferon Source) or the LEGEND MAXTM Mouse IFN-β ELISA Kit (BioLegend) following the manufacturer's protocol.

Techniques: Incubation, Concentration Assay, Cell Culture, Enzyme-linked Immunosorbent Assay

Journal: Biochemical Journal

Article Title: Suppression of interferon β gene transcription by inhibitors of bromodomain and extra-terminal (BET) family members

doi: 10.1042/BJ20141523

Figure Lengend Snippet: ( A and B ) Gen2.2 cells were incubated for 1 h with or without BI-2536 (1.0 μM), JQ1 (1.0 μM) or I-BET151 (1.0 μM) and then stimulated for 8 h with CL097 (1.0 μg/ml) ( A ) or for 12 h with ODN1826 (1.0 μM) ( B ). The concentration of IFNβ in the culture medium was measured by ELISA. Results are means+S.D. from two independent experiments each performed in duplicate. ( C ) Gen2.2 cells were incubated for 1 h with or without BI-2536 (1.0 μM), JQ1 (1.0 μM), I-BET151 (1.0 μM) or BI-605906 (5.0 μM) and then stimulated for 1 h with CL097. Staining with anti-IRF5, or DAPI to reveal nuclei, followed by deconvolution microscopy was performed as described in the Materials and methods section. ( D ) Gen2.2 cells were incubated for 1 h with BI-2536 (1 μM) or JQ1 (1 μM), then stimulated for 1 h with the TLR7 agonist CL097 (1 μg/ml), cross-linked and lysed. Chromatin was sheared by sonication and ChIP was performed using anti-IRF5. The enrichment of the Ifnb promoter was measured by qPCR, normalizing to input. Results are means+S.D. similar results were obtained in three independent experiments each performed in duplicate. IP, immunoprecipitation.

Article Snippet: The level of secreted IFNβ in the cell culture medium was determined using the Verikine mouse and human IFNβ ELISA kits (PBL Interferon Source) or the LEGEND MAXTM Mouse IFN-β ELISA Kit (BioLegend) following the manufacturer's protocol.

Techniques: Incubation, Concentration Assay, Enzyme-linked Immunosorbent Assay, Staining, Microscopy, Sonication, Immunoprecipitation

Journal: Emerging microbes & infections

Article Title: Novel transcription and replication-competent virus-like particles system modelling the Nipah virus life cycle.

doi: 10.1080/22221751.2024.2368217

Figure Lengend Snippet: Figure 6. Utility of trVLP-NiV in evaluating the effects of different kinds of antiviral agents. (A) 293T-NPL cells were infected with trVLP-NiV and cultured with 10 μM of FEMA 4809, remdesivir, and ribavirin, respectively. DMSO was used as a control. After 24 h infection, RLuc activity was quantified and normalized to DMSO-treated cells. Antiviral effect of ribavirin (B), remdesivir (C), IFN-α (D), and IFN-β (E). The 293T-NPL cells were infected with trVLP-NiV and treated with indicated concentrations of ribavirin or remdesivir. For IFN-α and IFN-β, cells were pre-treated with IFN-α or IFN-β for 8 h before infection. After 24 h of infection, RLuc activity (dark orange) and cell viability (blue) were measured. (F to H) Evaluating neutralizing activity with trVLP-NiV. The indicated concentrations of mAbs were incubated with trVLP-NiV for 1 h at 37 °C. In addition, residual infectivity was determined using 293T-NPL cells. IC50, the half-maximum inhibitory concentration; CC50, 50% cytotoxic concentration. N = 3. Error bars indi cate SEM. NS, no significance; *P < 0.05; **P < 0.01. Significance assessed by student’s t-test.

Article Snippet: For interferon-α (IFN-α) and IFN-β treatment, the cells were pre-treated with different concentrations of IFN-α (MCE, HYP73246) and IFN-β (Sino Biological, 10704-HNAS-5) 8 h prior to infection.

Techniques: Infection, Cell Culture, Control, Activity Assay, Incubation, Concentration Assay