Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Cutting edge: IL-1beta mediates the proangiogenic activity of osteopontin-activated human monocytes.

doi: 10.4049/jimmunol.177.7.4267

Figure Lengend Snippet: FIGURE 3. OPN-treated monocytes induce endothelial cell chemotaxis and angiogenesis in the CAM through IL-1. A, Murine aortic endothelial cells were assessed for their capacity to migrate in response to CM from control and OPN-treated monocytes in a Boyden chamber assay. After 4 h, cells that migrated through the filter were counted in triplicate (five fields per well) at 250 magnification. Control values obtained in fresh medium (52 8 cells/field) were subtracted from all the data. , Significantly different from control CM, p 0.01 by Student’s t test. B, Gelatin sponges were adsorbed with: vehicle (PBS); rFGF-2; CM from OPN-treated mono- cytes in the absence (CM) or in the presence of neutralizing anti-IL-1 (CM anti-IL-1), anti-IL-8 (CM anti-IL-8), or anti-TNF- (CM anti-TNF-) mAb; and rIL-1. Sponges were then implanted onto the CAM. At day 12, blood vessels entering the sponges were counted. Data are the mean SD of 20 embryos. , Significantly different from CM, p 0.05 by Student’s t test. C–E, Macroscopic images of CAM treated with CM from OPN-treated monocytes in the absence (C) or in the presence (D) of a neutralizing anti-IL-1 mAb. Note the significantly reduced angio- genic response in D, similar to that observed in the CAM treated with PBS (E). F and G, IL-1RI expression in the CAM. RT-PCR was performed using specific chicken IL-1RI primers on retrotranscribed CAM mRNA (RT). Nonretrotranscribed CAM mRNA ( RT) was used as control (F). Paraffin-embedded CAM sections were nuclear counterstained and deco- rated with affinity-purified goat polyclonal anti-IL-1RI Ab on day 12. IL- 1RI immunoreactivity was evident in the endothelium lining the blood vessels (G). Original magnification, 400. No specific signal was observed in CAM in which primary Ab was replaced by preimmune rabbit serum (data not shown).

Article Snippet: Immunohistochemistry and Western blot analysis IL-1RI immunodetection was performed on deparaffinized 5- m CAM sections (15), using a goat polyclonal anti-mouse IL-1RI Ab (R&D Systems) and a preimmune goat serum as negative control.

Techniques: Chemotaxis Assay, Control, Boyden Chamber Assay, Expressing, Reverse Transcription Polymerase Chain Reaction

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: IL-1β induces the rapid secretion of the antimicrobial protein IL-26 from Th17 cells

doi: 10.4049/jimmunol.1900318

Figure Lengend Snippet: (A) Sorting scheme used to isolate helper T cell subsets, a representative donor is shown, n = 3. (B) Isolated Th1, Th2, and Th17 cells were stimulated with IL-1β for 48 hours and cell free supernatants were analyzed for IL-26 secretion by ELISA, n = 3. (C) A representative sorting scheme for isolating IL-1RI+ and IL-1RI- Th17 cells, n= 3. (D, E) IL-1RI+ and IL-1RI- Th17 cells were stimulated with 100 ng/ml IL-1β or 5 µl/ml anti-CD3/CD28 for 9 hours before IL-26 (D) and IL-17A (E) was measured in cell free supernatants by ELISA, n = 3. (B,D) Data are represented as mean ± SEM, statistics calculated using one-way ANOVA with Tukey’s post-test (B, D, E). **p ≤ 0.01. *** p ≤ 0.001

Article Snippet: In some experiments, memory CD4 + T cells were incubated overnight with 1 nM recombinant human IL-2 and stained additionally with antibodies against APC-IL-1RI (R&D Systems, polyclonal goat anti-human) to sort IL-1RI + Th17 cells (CCR6 + CD161 + CCR4 + CXCR3 - IL-1RI + ) and IL-1RI - Th17 cells (CCR6 + CD161 + CCR4 + CXCR3 - IL-1RI - ).

Techniques: Isolation, Enzyme-linked Immunosorbent Assay

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: IL-1β induces the rapid secretion of the antimicrobial protein IL-26 from Th17 cells

doi: 10.4049/jimmunol.1900318

Figure Lengend Snippet: (A) Unsupervised principal component analysis of IL-1RI+ and IL-1RI- Th17 cells from 3 donors treated with IL-1β, anti-CD3/CD28, or media as a control. Numbers within symbols indicate the donor. (B) Hierarchical clustering of the samples. (C) Differential gene expression in untreated IL-1RI+ Th17 cells vs. IL-1RI- Th17 cells is represented in a volcano plot and some of the most differentially expressed genes are named and highlighted with black dots. (D) Genes with a (FC) ≥ 1.5 and p-adj ≤0.05 after treatment of IL-1RI+ Th17 cells with IL-1β vs media are plotted based on their normalized counts after IL-1β or anti-CD3/CD28 treatment. The line y = x was plotted on the graph to show genes with greater relative counts in each condition. Genes in the IPA “Th17 cell activation” canonical pathway and CXCL8 are highlighted with black dots. (E) Memory CD4+ T cells were treated with 100 ng/ml IL-1β and 5 µl/ml anti-CD3/CD28 for 48 hours before supernatants were collected and IL-8 protein concentrations were measured by ELISA, n = 4. Data are represented as mean ± SEM, statistics calculated using one-way ANOVA with Tukey’s post-test. *** p ≤ 0.001.

Article Snippet: In some experiments, memory CD4 + T cells were incubated overnight with 1 nM recombinant human IL-2 and stained additionally with antibodies against APC-IL-1RI (R&D Systems, polyclonal goat anti-human) to sort IL-1RI + Th17 cells (CCR6 + CD161 + CCR4 + CXCR3 - IL-1RI + ) and IL-1RI - Th17 cells (CCR6 + CD161 + CCR4 + CXCR3 - IL-1RI - ).

Techniques: Control, Gene Expression, Activation Assay, Enzyme-linked Immunosorbent Assay

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: IL-1β induces the rapid secretion of the antimicrobial protein IL-26 from Th17 cells

doi: 10.4049/jimmunol.1900318

Figure Lengend Snippet: “Th17 cell activation” pathway genes significantly upregulated by IL-1β treatment of IL-1RI + Th17 cells. The IPA canonical pathway “Th17 cell activation” was the most significantly enriched canonical pathway within our dataset (p = 2.51 × 10 −17 ). This pathway contained 90 genes, 19 of which, or 21%, were contained in our dataset and are displayed in the table.

Article Snippet: In some experiments, memory CD4 + T cells were incubated overnight with 1 nM recombinant human IL-2 and stained additionally with antibodies against APC-IL-1RI (R&D Systems, polyclonal goat anti-human) to sort IL-1RI + Th17 cells (CCR6 + CD161 + CCR4 + CXCR3 - IL-1RI + ) and IL-1RI - Th17 cells (CCR6 + CD161 + CCR4 + CXCR3 - IL-1RI - ).

Techniques: Activation Assay

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: IL-1β induces the rapid secretion of the antimicrobial protein IL-26 from Th17 cells

doi: 10.4049/jimmunol.1900318

Figure Lengend Snippet: (A, B) Genes with a log2(FC)≥ 0.58 and p-adj ≤0.05 after treatment of IL-1RI+ Th17 cells with IL-1β (A) or anti-CD3/CD28 (B) were analyzed for enrichment of upstream transcription factors using Enrichr and plotted with their -log10(p-adj). C. Memory CD4+ T cells were stimulated with either 20 ng/ml IL-1β or 5 µl/ml anti-CD3/CD28 for 24 hours, with or without 1 hour pretreatment with 10µM Bay 11–7082, an NF-κB activation inhibitor. IL-26 secretion into cell supernatants was measured by ELISA, n = 3. Data are represented as mean ± SEM, statistics calculated using one-way ANOVA with Tukey’s post-test. *p ≤ 0.05, *** p ≤ 0.001.

Article Snippet: In some experiments, memory CD4 + T cells were incubated overnight with 1 nM recombinant human IL-2 and stained additionally with antibodies against APC-IL-1RI (R&D Systems, polyclonal goat anti-human) to sort IL-1RI + Th17 cells (CCR6 + CD161 + CCR4 + CXCR3 - IL-1RI + ) and IL-1RI - Th17 cells (CCR6 + CD161 + CCR4 + CXCR3 - IL-1RI - ).

Techniques: Activation Assay, Enzyme-linked Immunosorbent Assay

Journal: Cell death & disease

Article Title: Human proximal tubular epithelial cell interleukin-1 receptor signalling triggers G2/M arrest and cellular senescence during hypoxic kidney injury.

doi: 10.1038/s41419-025-07386-6

Figure Lengend Snippet: Fig. 4 Significantly elevated PTEC IL-1RI expression and senescence in fibrotic kidneys. A Left panel: Quantitative analysis of % IL-1RI+ PTECs (proportion of AQP-1+ cells (i.e., PTECs) expressing IL-1RI) in control (non-fibrotic; n = 4) and fibrotic kidney tissue (n = 5). Points represent values for individual donor tissue samples. Results represent mean ± SEM of values from four randomly selected areas for each tissue sample. *P < 0.05, Welch’s t-test. Right panel: Representative immunofluorescent labelling of control (non-fibrotic) and fibrotic kidney tissue stained for PTEC marker aquaporin-1 (AQP-1; white), IL-1RI (green) and DAPI (blue). Scale bars represent 20 µm. IL-1RI+ PTECs are highlighted with white arrows. B Left panel: Quantitative analysis of % SA-β-gal+ PTECs (proportion of AQP-1+ cells (i.e., PTEC) with positive Spider-β-gal staining) in control (non-fibrotic; n = 4) and fibrotic kidney tissue (n = 4). Points represent values for individual donor tissue samples. Results represent mean ± SEM of values from four randomly selected areas for each tissue sample. **P < 0.01, Welch’s t-test. Right panel: Representative immunofluorescent images of control (non-fibrotic) and fibrotic kidney tissue labelled with PTEC marker AQP-1 (white), Spider-β-gal (green) and DAPI (blue). Scale bars represent 20 µm. SA-β-gal+ PTECs are highlighted with white arrows. Lower magnification immunofluorescent images are presented in Fig. S4.

Article Snippet: Sections were subsequently probed with primary antibodies against IL-1RI (1:70; Goat polyclonal IgG; Cat. No. AF269SP; R&D Systems, Minneapolis, MN, USA) and PTEC marker aquaporin-1 (AQP-1; 1:50; Mouse monoclonal IgG; Clone B-11; Cat. No. sc-25287; Santa Cruz, Dallas, TX, USA) overnight at 4 °C in a humidified chamber.

Techniques: Expressing, Control, Staining, Marker

Journal: Reproduction (Cambridge, England)

Article Title: Increased expression of interleukin-1 receptor type 1 in active endometriotic lesions.

doi: 10.1530/rep.1.01121

Figure Lengend Snippet: Figure 1 Immunohistochemical analysis of IL-1RI expression in endometriotic tissue. (A) Note the intense brown immunostaining in the glands and cell aggregates throughout the stroma in the presence of a polyclonal goat anti-human IL-1RI antibody (a) and the absence of such staining in the presence of goat IgGs used at concentration equivalent to that of the primary antibody (b; negative control); scale bar, 30 mm. (B) HSCORE in red (nZ7), typical (nZ6), and white (nZ5) endometriotic lesions. G, gland, S, stroma; *, † and ‡significant difference with red lesions (P!0.05, !0.01, and !0.001 respectively) as analyzed by ANOVA and the Bonferroni’s test for multiple comparisons.

Article Snippet: This assay uses a mouse monoclonal anti-human IL-1RI antibody (capture antibody) that does not cross-react with rhIL1RII (R & D Systems) and a goat polyclonal anti-human IL-1RI antibody (detection antibody) that shows 5–10% Reproduction (2007) 133 265–274 cross-reactivity with rhIL-1RII (R&D Systems).

Techniques: Immunohistochemical staining, Expressing, Immunostaining, Staining, Concentration Assay, Negative Control

Journal: Reproduction (Cambridge, England)

Article Title: Increased expression of interleukin-1 receptor type 1 in active endometriotic lesions.

doi: 10.1530/rep.1.01121

Figure Lengend Snippet: Figure 2 Dual-immunofluorescent staining of IL-1RI (A–C) and CD3 (D), CD68 (E), or vWF (F) in endometriotic tissue. Tissue sections were incubated with goat polyclonal anti- human IL-1RI antibody and with mouse monoclonal anti-CD3, mouse monoclonal CD68, or mouse monoclonal anti-human vWF antibody. Sections were then incubated simultaneously with fluorescein isothiocya- nate-conjugated donkey anti-goat antibody and rhodamine-conjugated sheep anti-mouse antibody to detect co-expression of IL-1RI with CD3, CD68, or vWF. Note the expression of IL-1RI (green) in CD3-, CD68-, and vWF- positive T-lymphocytes, macrophages, and endothelial cells respectively (red). Super- position of fluorescein (green) and rhodamine (red) signals clearly shows co-expression (yellow signal) of IL-1RI with CD3 (GZACD), CD68 (HZBCE), and vWF (IZCCF). Scale bars, 30 mm.

Article Snippet: This assay uses a mouse monoclonal anti-human IL-1RI antibody (capture antibody) that does not cross-react with rhIL1RII (R & D Systems) and a goat polyclonal anti-human IL-1RI antibody (detection antibody) that shows 5–10% Reproduction (2007) 133 265–274 cross-reactivity with rhIL-1RII (R&D Systems).

Techniques: Staining, Incubation, Expressing