il1ra  (Proteintech)

Journal: Journal of Clinical and Translational Hepatology

Article Title: IL1RA + Myeloid-derived Suppressor Cells Activate Epithelial-mesenchymal Transition to Facilitate Lymphatic and Hepatic Metastasis in Pancreatic Ductal Carcinoma

doi: 10.14218/JCTH.2025.00416

Figure Lengend Snippet: (A) tSNE plot showing the four neutrophil-derived MDSC subtypes. (B) The top five expressed genes of IL1RN + MDSCs in the four neutrophil-derived MDSC subsets. (C, D) Representative flow cytometry data and quantification graphs showing the proportion of IL1RA + MDSCs in LNN and LNP PDAC tissues. (E) Representative images of immunofluorescence staining of IL1RA + MDSCs in LNN and LNP PDAC tissues. Scale bar = 100 µm. (F) Trajectory of neutrophil-derived MDSCs along pseudotime in two-dimensional space. (G) Heatmap showing dynamic changes in gene expression along pseudotime. (H) Prognostic analysis of PDAC patients with low or high IL1RA + MDSC recruitment (stratified by the median IL1RA + MDSC proportion in all samples). Kaplan–Meier survival plots of 16 patients are shown. Data are presented as mean ± SD in (D). Statistical differences were assessed using a two-tailed Student’s t -test. LNN, lymph node-negative; LNP, lymph node-positive; PDAC, pancreatic ductal adenocarcinoma; MDSCs, myeloid-derived suppressor cells.

Article Snippet: Primary antibodies included CD11b (Abcam, Cambridge, UK), CD68 (Proteintech, Wuhan, China), MPO (Abcam), IL1RA (Proteintech), and VEGFA (Proteintech).

Techniques: Derivative Assay, Flow Cytometry, Immunofluorescence, Staining, Gene Expression, Two Tailed Test

Journal: Journal of Clinical and Translational Hepatology

Article Title: IL1RA + Myeloid-derived Suppressor Cells Activate Epithelial-mesenchymal Transition to Facilitate Lymphatic and Hepatic Metastasis in Pancreatic Ductal Carcinoma

doi: 10.14218/JCTH.2025.00416

Figure Lengend Snippet: (A) Schematic illustration of IL1RN + MDSC–PDAC co-culture system. (B) Transwell migration assays in PANC-1 and ASPC-1 cells treated with normal medium (Control), IL1RA + MDSC-conditioned medium (IL1RA + MDSC CM), or IL1RA + MDSC CM + Axitinib. Scale bar = 50 µm. (C) Western blot analysis of EMT markers (N-cadherin, E-cadherin, ZO-1, Claudin-1) in PANC-1 cells under treatments identical to (B); β-actin was detected as a control. Grayscale was determined using ImageJ. (D) Relative expression of stemness-related mRNA after treatment with IL1RA + MDSC CM or IL1RA + MDSC CM + Axitinib. (E) Representative images of subcutaneous tumors in BALB/c nude mice from the PDX model: Group 1 (Control, tumor tissue only); Group 2 (tumor tissue co-implanted with IL1RN + MDSCs); Group 3 (tumor tissue co-implanted with IL1RN + MDSCs + Axitinib). (F) Excised tumor tissues from (E). (G, H) Tumor weights (G) and volumes (H) from the mouse model presented as bar graphs. (I) Representative H&E staining images of mouse pancreatic tumor tissues. Scale bar = 100 µm. Data are represented as mean ± SD of three independent biological replicates in (B) and four independent replicates in (F and G). Statistical significance was determined by one-way ANOVA with Dunnett’s post-hoc test: ns p ≥ 0.05; * p < 0.05; *** p < 0.001; **** p < 0.0001. PDAC, pancreatic ductal adenocarcinoma; MDSCs, myeloid-derived suppressor cells.

Article Snippet: Primary antibodies included CD11b (Abcam, Cambridge, UK), CD68 (Proteintech, Wuhan, China), MPO (Abcam), IL1RA (Proteintech), and VEGFA (Proteintech).

Techniques: Co-Culture Assay, Migration, Control, Western Blot, Expressing, Staining, Derivative Assay

Journal: ImmunoHorizons

Article Title: Deficiency of interleukin-1 receptor antagonist aggravates Citrobacter rodentium infection in mice

doi: 10.1093/immhor/vlaf034

Figure Lengend Snippet: IL1ra KO mice display elevated basal inflammatory tone and leukocytosis. IL1ra KO mice and WT littermates (8-wk-old female, n = 5) were assessed for circulating (A) white blood cells (WBCs), (B) lymphocytes, and (C) neutrophils via Abaxis VetScan HM5 hematology analyzer. (D, E) Splenic and (F, G) colonic lamina propria CD11b + Ly6G + neutrophils were quantified using flow cytometry. (H) SAA, (I) serum Lcn2, and (J) fecal Lcn2 were measured by ELISA. (K) Representative images of hematoxylin and eosin (H&E) colon sections (scale bar = 100 µm). Results are representative of 2 independent experiments, and are expressed as mean ± SEM. Student’s t test. * P < 0.05, ** P < 0.01.

Article Snippet: WT (stock #000664) and IL1ra KO (stock #004754) mice on C57BL/6J background were procured from the Jackson Laboratory and crossed to generate WT and IL1ra KO littermates.

Techniques: Flow Cytometry, Enzyme-linked Immunosorbent Assay

Journal: ImmunoHorizons

Article Title: Deficiency of interleukin-1 receptor antagonist aggravates Citrobacter rodentium infection in mice

doi: 10.1093/immhor/vlaf034

Figure Lengend Snippet: IL-1RA deficiency aggravates C. rodentium ( CR ) infection. IL1ra KO mice and WT littermates (8-wk-old female, n = 5) were orally infected with 1 × 10 9 CFU of CR (strain ICC169, nalidixic acid resistant) and euthanized on day 11 postinfection. (A) Body weight. Bacterial burden of CR was determined by CFU in (B) feces, (C) cecal content, (D) mesenteric lymph nodes (MLNs), (E) spleen, and (F) liver. (G) Representative image depicts gross colon and spleen. (H) Colon and (I) spleen weight (shown as percent of body weight). (J) Fecal LCN2, (K) serum LCN2, and (L) SAA were measured via ELISA. (M) Histological scoring of colon pathology. Paraffin-embedded colon sections stained with (N) hematoxylin and eosin (H&E) (scale bar = 200 µm) and (O) Alcian blue (scale bar = 200 µm). Results are representative of two independent experiments, and expressed as means ± SEM. Student’s t test. * P < 0.05. Con, control.

Article Snippet: WT (stock #000664) and IL1ra KO (stock #004754) mice on C57BL/6J background were procured from the Jackson Laboratory and crossed to generate WT and IL1ra KO littermates.

Techniques: Infection, Enzyme-linked Immunosorbent Assay, Staining, Control

Journal: ImmunoHorizons

Article Title: Deficiency of interleukin-1 receptor antagonist aggravates Citrobacter rodentium infection in mice

doi: 10.1093/immhor/vlaf034

Figure Lengend Snippet: Absence of IL-1RA increases colonic pathogen load during C. rodentium ( CR ) infection. IL1ra KO mice and WT littermates (8-wk-old male, n = 5) were orally infected with 1 × 10 9 CFU of bioluminescent CR (strain ICC180, kanamycin-resistant) and euthanized on day 15 postinfection. (A) Representative images depicted mice imaged for bioluminescence using the Xenogen IVIS Spectrum. (B) Bioluminescent counts. (C) Bacterial burden of CR was determined by plating fecal homogenate on kanamycin-containing LB agar plates and quantified as CFU. (D) Fecal LCN2 was measured using ELISA. Results presented as means ± SEM. Student’s t test. * P < 0.05, ** P < 0.01.

Article Snippet: WT (stock #000664) and IL1ra KO (stock #004754) mice on C57BL/6J background were procured from the Jackson Laboratory and crossed to generate WT and IL1ra KO littermates.

Techniques: Infection, Enzyme-linked Immunosorbent Assay

Journal: ImmunoHorizons

Article Title: Deficiency of interleukin-1 receptor antagonist aggravates Citrobacter rodentium infection in mice

doi: 10.1093/immhor/vlaf034

Figure Lengend Snippet: IL-1RA deficiency heightens neutrophil function, but does not impair its ability to kill C. rodentium in vitro. (A) Representative immunohistochemical staining of MPO (a neutrophil marker) on colon sections from WT and IL1ra KO mice (8-wk-old male, n = 5) that were infected with C. rodentium (related to Fig. 1 ). (B) Colonic MPO was measured using ELISA. (C) BMDNs were isolated from WT and IL1ra KO mice (8-wk-old male, n = 4), stimulated with PBS (Con), LPS, PMA, or calcium ionophore and then measured for the formation of NETs ex vivo. The data were presented as fold change of NETs formation normalized to the controls. (D) Another group of BMDNs were stimulated with PBS, LPS, or PMA and then measured for the production of ROS. Neutrophil ROS was expressed as fold change of mean fluorescence intensity (MFI) normalized to the controls. WT and IL1ra KO BMDNs (5 × 10 5 cells) were incubated with C. rodentium (1 × 10 5 CFU) and then quantified for bacterial survival in the (E) culture supernatant and (F) cell lysates. Results presented as means ± SEM. Student’s t test. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: WT (stock #000664) and IL1ra KO (stock #004754) mice on C57BL/6J background were procured from the Jackson Laboratory and crossed to generate WT and IL1ra KO littermates.

Techniques: In Vitro, Immunohistochemical staining, Staining, Marker, Infection, Enzyme-linked Immunosorbent Assay, Isolation, Ex Vivo, Fluorescence, Incubation

Journal: ImmunoHorizons

Article Title: Deficiency of interleukin-1 receptor antagonist aggravates Citrobacter rodentium infection in mice

doi: 10.1093/immhor/vlaf034

Figure Lengend Snippet: IL-1RA deficiency alters macrophage function and impairs its ability to kill C. rodentium in vitro. BMDMs were isolated from WT and IL1ra KO mice (8-wk-old male, n = 4), and stimulated with PBS (Con), LPS, or flagellin (FliC). BMDM culture supernatant was measured for (A) IL-1RA and (B) IL-1β using ELISA. (C) Immunoblot of inducible nitric oxide synthase (iNOS) in the BMDM cell lysates, and (D) its quantification using densitometry scanning. (E) Nitrite secretion from the BMDMs culture was measured using Griess reagent. WT and IL1ra KO BMDMs (5 × 10 5 cells) were incubated with C. rodentium (1 × 10 5 CFU) and then quantified for bacterial survival in (E) culture supernatant and (F) cell lysates. Results presented as means ± SEM. Data in panels A, B, D, and E were analyzed via 1-way analysis of variance with Tukey’s multiple comparison tests, whereas data in panels F and G were analyzed via Student’s t test. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: WT (stock #000664) and IL1ra KO (stock #004754) mice on C57BL/6J background were procured from the Jackson Laboratory and crossed to generate WT and IL1ra KO littermates.

Techniques: In Vitro, Isolation, Enzyme-linked Immunosorbent Assay, Western Blot, Incubation, Comparison