recombinant il 33 ril 33  (R&D Systems)

Journal: Inflammation

Article Title: Interleukin-33 Promotes Neutrophil Extracellular Trap Formation To Aggravate Renal Ischemia-Reperfusion Injury Through ST2/PI3K/Akt and ST2/PAD4 Pathways

doi: 10.1007/s10753-025-02364-8

Figure Lengend Snippet: IL-33 was upregulated and positively correlated with NET formation in renal transplant patients. ( A ) Paired analysis of serum IL-33 levels, ( B ) paired analysis of serum MPO-DNA levels, ( C ) paired analysis of serum citH3 levels in renal transplant patients preoperatively and postoperatively ( n = 20). ( D ) Correlation analysis for postoperative serum IL-33 level and serum MPO-DNA level (Spearman r = 0.6892, P = 0.0008). ( E ) Correlation analysis of postoperative serum IL-33 level and serum citH3 level (Spearman r = 0.6486, P = 0.002). * P < 0.05, ** P < 0.01

Article Snippet: Mice were intraperitoneally injected with recombinant IL-33 (rIL-33) (10ug per mouse, MedChemExpress, America) or PBS immediately following exposure to renal ischemia.

Techniques:

Journal: Inflammation

Article Title: Interleukin-33 Promotes Neutrophil Extracellular Trap Formation To Aggravate Renal Ischemia-Reperfusion Injury Through ST2/PI3K/Akt and ST2/PAD4 Pathways

doi: 10.1007/s10753-025-02364-8

Figure Lengend Snippet: IL-33 and NETs were elevated following renal I/R in mice. ( A - B ) Serum IL-33 levels ( A ) and renal tissue homogenate IL-33 levels ( B ) after renal I/R ( n = 6). ( C - D ) Representative blots ( C ) and statistical analysis ( D ) of IL-33 protein expression levels in renal tissues after I/R ( n = 6). ( E - F ) Representative images ( E ) and statistical analysis ( F ) of IL-33 immunohistochemistry in renal tissues after I/R ( n = 6). Scale bar was 50 μm. ( G - H ) IL-33 expression and distribution among the groups (blue staining indicate DAPI for nuclei), IL-33 (red) and CD31 (green) ( n = 6). Scale bar was 50 μm. (I-J) Serum MPO-DNA (I) and citH3 levels ( J ) following renal I/R ( n = 6). ( K - L ) Representative blots ( K ) and statistical analysis ( L ) of citH3 protein expression levels in renal tissues after I/R ( n = 6). ( M - N ) NET formation in in the indicated groups (blue indicate DAPI staining), MPO (red) and citH3 (green) ( n = 6). Scale bar was 20 μm. ns no significance, * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: Mice were intraperitoneally injected with recombinant IL-33 (rIL-33) (10ug per mouse, MedChemExpress, America) or PBS immediately following exposure to renal ischemia.

Techniques: Expressing, Immunohistochemistry, Staining

Journal: Inflammation

Article Title: Interleukin-33 Promotes Neutrophil Extracellular Trap Formation To Aggravate Renal Ischemia-Reperfusion Injury Through ST2/PI3K/Akt and ST2/PAD4 Pathways

doi: 10.1007/s10753-025-02364-8

Figure Lengend Snippet: IL-33 promoted NET generation during renal IRI. ( A ) A diagram showing WT mice receiving renal IRI or sham surgery following intraperitoneal injection of rIL-33 (10 µg per mouse) or vehicle (PBS). ( B - C ) Serum MPO-DNA ( B ) and citH3 levels ( C ) of mice in each group ( n = 6). ( D - E ) Representative blots ( D ) and statistical analyses ( E ) of citH3 protein expression levels in renal tissues in the indicated groups ( n = 6). ( F - G ) NET formation in the indicated groups through DAPI staining (blue), MPO (red) and citH3 (green) ( n = 6). Scale bar was 20 μm. ( H - I ) Serum Cr ( H ) and BUN levels ( I ) in the indicated groups ( n = 6). ( J - K ) Representative blots ( J ) and statistical analysis ( K ) of KIM-1 protein expression levels in renal tissues in the indicated groups ( n = 6). ( L - N ) Renal tissues from mice in the indicated groups stained with HE and KIM-1 ( L ), followed by score of tubular injury ( M ) and quantitative analysis of KIM-1 ( N ) ( n = 6). Scale bar was 50 μm. * P < 0.05, ** P < 0.01

Article Snippet: Mice were intraperitoneally injected with recombinant IL-33 (rIL-33) (10ug per mouse, MedChemExpress, America) or PBS immediately following exposure to renal ischemia.

Techniques: Injection, Expressing, Staining

Journal: Inflammation

Article Title: Interleukin-33 Promotes Neutrophil Extracellular Trap Formation To Aggravate Renal Ischemia-Reperfusion Injury Through ST2/PI3K/Akt and ST2/PAD4 Pathways

doi: 10.1007/s10753-025-02364-8

Figure Lengend Snippet: IL-33 exacerbated renal IRI by inducing NET formation. ( A ) A diagram showing the time node of intraperitoneal injection of GSK484 (4 mg/kg) and rIL-33 (10 µg per mouse) in WT mice before renal I/R. (B-C) Serum MPO-DNA ( B ) and citH3 levels ( C ) in the indicated groups ( n = 6). ( D - E ) Representative blots ( D ) and statistical analysis ( E ) of citH3 expression levels in renal tissues of the indicated groups ( n = 6). ( F - G ) NET formation in the indicated groups as determined using the DAPI (blue), MPO (red) and citH3 (green) staining ( n = 6). Scale bar was 20 μm. ( H - I ) Serum Cr ( H ) and BUN levels ( I ) in the indicated groups ( n = 6). ( J - K ) Representative blots ( J ) and statistical analysis ( K ) of KIM-1 protein expression levels in renal tissues from the indicated groups ( n = 6). ( L - N ) Renal tissues of mice in the indicated groups stained with HE and KIM-1 ( L ), followed by score of tubular injury ( M ) and quantitative analysis of KIM-1 ( N ) ( n = 6). Scale bar was 50 μm. ** P < 0.01, *** P < 0.001

Article Snippet: Mice were intraperitoneally injected with recombinant IL-33 (rIL-33) (10ug per mouse, MedChemExpress, America) or PBS immediately following exposure to renal ischemia.

Techniques: Injection, Expressing, Staining

Journal: Inflammation

Article Title: Interleukin-33 Promotes Neutrophil Extracellular Trap Formation To Aggravate Renal Ischemia-Reperfusion Injury Through ST2/PI3K/Akt and ST2/PAD4 Pathways

doi: 10.1007/s10753-025-02364-8

Figure Lengend Snippet: Blocking IL-33 improved renal IRI by reducing NET formation. ( A ) The diagram of WT mice receiving renal IRI or sham surgery after intraperitoneal injection of anti-IL-33 monoclonal antibody (Anti-IL-33) (10 µg per ounce) or vehicle (IgG2b Isotype Control). ( B - C ) Serum MPO-DNA ( B ) and citH3 levels ( C ) of mice in the indicated groups ( n = 6). (D-E) Representative blots ( D ) and statistical analysis ( E ) of citH3 protein expression levels in renal tissues in the indicated groups ( n = 6). ( F - G ) NET formation in the indicated groups following DAPI (blue), MPO (red) and citH3 (green) staining ( n = 6). Scale bar was 20 μm. ( H - I ) Serum Cr ( H ) and BUN levels ( I ) in the indicated group ( n = 6). ( J - K ) Representative blots ( J ) and statistical analysis ( K ) of KIM-1 protein expression levels in renal tissues in the indicated groups ( n = 6). ( L - N ) Renal tissues of mice in the indicated groups subjected to HE and KIM-1 staining ( L ), followed by score of tubular injury ( M ) and statistical analysis of KIM-1 ( N ) ( n = 6). Scale bar was 50 μm. ** P < 0.01, *** P < 0.001

Article Snippet: Mice were intraperitoneally injected with recombinant IL-33 (rIL-33) (10ug per mouse, MedChemExpress, America) or PBS immediately following exposure to renal ischemia.

Techniques: Blocking Assay, Injection, Control, Expressing, Staining

Journal: Inflammation

Article Title: Interleukin-33 Promotes Neutrophil Extracellular Trap Formation To Aggravate Renal Ischemia-Reperfusion Injury Through ST2/PI3K/Akt and ST2/PAD4 Pathways

doi: 10.1007/s10753-025-02364-8

Figure Lengend Snippet: IL-33 stimulated NET generation in vitro. ( A - B ) Neutrophils incubated with PBS, PMA (100nM) and various concentrations of rIL-33 (20, 60, 100 ng/mL) for 4 h. rIL-33 stimulation increased MPO-DNA ( A ) and citH3 levels ( B ) in the neutrophil culture medium in a dose-dependent manner relative to the PBS group ( n = 3). ( C - D ) Relative to the PBS group, rIL-33 activated neutrophils to increase citH3 protein in a dose-dependent manner ( n = 3). ( E ) Compared with the PBS group, rIL-33 increased the expression of ST2 mRNA on neutrophils ( n = 3). ( F ) Representative scanning electron microscopy graphs of neutrophils treated with PBS or rIL-33 (100ng/mL) for 4 h. Scale bar was 10 μm. ( G ) The NET formation as indicated by DAPI (blue), MPO (red) and citH3 (green) staining, was comparable between rIL-33 (100ng/mL) and PMA group. Scale bar was 50 μm. ns no significance, * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: Mice were intraperitoneally injected with recombinant IL-33 (rIL-33) (10ug per mouse, MedChemExpress, America) or PBS immediately following exposure to renal ischemia.

Techniques: In Vitro, Incubation, Expressing, Electron Microscopy, Staining

Journal: Inflammation

Article Title: Interleukin-33 Promotes Neutrophil Extracellular Trap Formation To Aggravate Renal Ischemia-Reperfusion Injury Through ST2/PI3K/Akt and ST2/PAD4 Pathways

doi: 10.1007/s10753-025-02364-8

Figure Lengend Snippet: IL-33 induced NET formation via ST2/PI3K/Akt and ST2/PAD4 signaling pathways. Mouse bone marrow-derived neutrophils were treated with PBS or rIL-33 (100ng/mL) for 4 h, followed by RNA sequencing ( n = 3). ( A - B ) Volcano plot ( A ) and heatmap ( B ) showing differential gene expression between PBS and rIL-33 groups. ( C ) GO enrichment analysis of DEGs. ( D ) KEGG pathway enrichment analysis of DEGs. Bone marrow-derived neutrophils from WT and ST2 KO mice were treated with PBS or rIL-33 (100ng/mL) for 4 h. The production of MPO-DNA ( E ) and citH3 ( F ) in the cell culture medium of ST2 KO neutrophils treated with IL-33 was markedly decreased relative to the IL-33-treated WT neutrophils ( n = 3). ( G ) Confocal microscopy was conducted to examine NET formation (co-localization of DAPI, MPO and citH3) in each group. ( H - I ) The expression of PI3K, p-PI3K, Akt, p-Akt and citH3 in WT and ST2 KO neutrophils stimulated by rIL-33 as determined by Western blot ( n = 3). ( J - K ) WT neutrophils were treated with 10µM LY294002 (PI3K inhibitor) or 10µM MK2206 (Akt inhibitor) for 24 h, and 100ng/mL rIL-33 was added on the 20th h to stimulate WT neutrophils for 4 h. The expression of PI3K, p-PI3K, Akt, p-Akt and citH3 in neutrophils was determined by Western blot ( n = 3). ( L ) Gene expression of Padi1, Padi2, Padi4 and Padi6 in neutrophils stimulated with rIL-33 ( n = 3). The gene expression data were expressed as log 2 (FPKM + 1). ( M - N ) PAD4 protein expression levels and quantitative analysis in WT and ST2 KO neutrophils treated with rIL-33 were quantified by Western blots ( n = 3). ( O - P ) WT neutrophils were pretreated with GSK484 (PAD4 inhibitor) for 30 min after treatment with 100ng/mL rIL-33 for 4 h. The protein expression of citH3 in neutrophils was determined by Western blot ( n = 3). ns no significance, * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: Mice were intraperitoneally injected with recombinant IL-33 (rIL-33) (10ug per mouse, MedChemExpress, America) or PBS immediately following exposure to renal ischemia.

Techniques: Protein-Protein interactions, Derivative Assay, RNA Sequencing, Gene Expression, Cell Culture, Confocal Microscopy, Expressing, Western Blot

Journal: Inflammation

Article Title: Interleukin-33 Promotes Neutrophil Extracellular Trap Formation To Aggravate Renal Ischemia-Reperfusion Injury Through ST2/PI3K/Akt and ST2/PAD4 Pathways

doi: 10.1007/s10753-025-02364-8

Figure Lengend Snippet: Schematic diagram of the mechanism by which IL-33 exacerbated renal IRI by enhancing NET formation during renal I/R

Article Snippet: Mice were intraperitoneally injected with recombinant IL-33 (rIL-33) (10ug per mouse, MedChemExpress, America) or PBS immediately following exposure to renal ischemia.

Techniques:

Journal: Immunology

Article Title: Interleukin‐33 alleviates psoriatic inflammation by suppressing the T helper type 17 immune response

doi: 10.1111/imm.13203

Figure Lengend Snippet: Interleukin‐33 (IL‐33) expression is markedly increased in individuals with moderate‐to‐severe psoriasis. (a) mRNA expression of IL‐33 in the skin of normal people (n = 16) and lesional skin of psoriasis patients (n = 16). (b) mRNA expression of IL‐17A in the skin of normal people (n = 6) and lesional skin of psoriasis patients (n = 6). (c) The correlation of the mRNA expression of IL‐33 and IL‐17A in the skin (n = 12). (d,e) Immunohistochemical staining of IL‐33 in the epidermis (d) and dermis (e) of skin of normal people (n = 10) and lesional skin of psoriasis patients (n = 10). (f) Serum level of IL‐33 of normal people (n = 15) and psoriasis patients (n = 25). (g) mRNA expression of IL‐33 and IL‐17A in the lesional of psoriasis patients before and after anti‐tumor necrosis factor‐α (TNF‐α) therapy (n = 5). (h) Serum level of IL‐33 of psoriasis patients before and after anti‐TNF‐α therapy (n = 10). Data show mean + SEM or mean ± SEM. P‐values were determined by unpaired Student’s t‐test. Correlation of the mRNA expression of IL‐33 and IL‐17A was determined by Pearson coefficient. **P < 0·01, ***P < 0·001 and ****P < 0·0001

Article Snippet: Mouse recombinant IL‐33 (rIL‐33, 1 µg in 300 μl PBS; PeproTech, Rocky Hill, NJ) or PBS was injected subcutaneously daily into the back skin of the mice.

Techniques: Expressing, Immunohistochemical staining, Staining

Journal: Immunology

Article Title: Interleukin‐33 alleviates psoriatic inflammation by suppressing the T helper type 17 immune response

doi: 10.1111/imm.13203

Figure Lengend Snippet: Interleukin‐33 (IL‐33) suppresses T helper type 17 (Th17) cells in individuals with moderate‐to‐severe psoriasis. (a,b) The proportion of IL‐17A+ IFN‐γ –, IL‐17A+ IFN‐γ + and IL‐17A– IFN‐γ + cells and the geometric mean fluorescence intensity (gMFI) of IL‐17A and interferon‐γ (IFN‐γ) in CD4+ T cells from psoriasis patients (n = 17) treated with or without IL‐33 (50 ng/ml) for 72 hr. (c) The supernatant level of IL‐4 in CD4+ T cells from psoriasis patients (n = 10) treated with or without IL‐33 (50 ng/ml) for 72 hr. (d) The proportion of IL‐10+ and IFN‐γ + cells in Th17 (CD4+ and IL‐17A+) cells from psoriasis patients (n = 10) treated with or without IL‐33 (50 ng/ml) for 72 hr. (e) The proportion of regulatory T (Treg) (CD25+ and Foxp3+) cells in CD4+ T cells from psoriasis patients (n = 10) treated with or without IL‐33 (50 ng/ml) for 72 hr. (f) The proportion of invariant natural killer T (iNKT) (CD3+ and αGalCer‐CD1d+) cells in the peripheral blood mononuclear cells (PBMCs) from psoriasis patients (n = 10) treated with αGalCer and with or without IL‐33 (50 ng/ml) for 72 hr. Data show mean + SEM. P‐values were determined by paired Student’s t‐test. ns, no significance, *P < 0·05, **P < 0·01, ***P < 0.001 and ****P < 0.0001

Article Snippet: Mouse recombinant IL‐33 (rIL‐33, 1 µg in 300 μl PBS; PeproTech, Rocky Hill, NJ) or PBS was injected subcutaneously daily into the back skin of the mice.

Techniques: Fluorescence

Journal: Immunology

Article Title: Interleukin‐33 alleviates psoriatic inflammation by suppressing the T helper type 17 immune response

doi: 10.1111/imm.13203

Figure Lengend Snippet: Subcutaneous injection of interleukin‐33 (IL‐33) alleviates imiquimod (IMQ) ‐induced murine psoriatic inflammation. (a) Schematic representation of the plan with the injection of PBS or IL‐33 on mice treated with IMQ. (b) Representative photos of the lesional skin of mice from each group. (c) The disease severity scoring of the mice from each group based on scaling, erythema, and skin thickness from day 1 to day 7. (d) Representative hematoxylin & eosin staining of skin sections of mice from each group on day 7 (bar = 200 um). (e) Epidermal thickness and infiltrating inflammatory cells of the skin sections of mice from each group on day 7. Each group contains six mice and all experiments were repeated at least twice. Data show mean + SEM. P‐values were determined by one‐way analysis of variane with Bonferroni multiple‐comparisons t‐test. *P < 0·05, **P < 0·01 and ***P < 0·001

Article Snippet: Mouse recombinant IL‐33 (rIL‐33, 1 µg in 300 μl PBS; PeproTech, Rocky Hill, NJ) or PBS was injected subcutaneously daily into the back skin of the mice.

Techniques: Injection, Staining

Journal: Immunology

Article Title: Interleukin‐33 alleviates psoriatic inflammation by suppressing the T helper type 17 immune response

doi: 10.1111/imm.13203

Figure Lengend Snippet: Interleukin‐33 (IL‐33) altered the expression of inflammatory cytokines and T‐cell subpopulations of skin‐draining lymph nodes in imiquimod (IMQ) ‐treated mice. (a) The protein level of tumor necrosis factor‐α (TNF‐α), IL‐23, IL‐10 and interferon‐γ (IFN‐γ) of the skin of mice from each group. (b) The serum level of TNF‐α and IFN‐γ of the mice from each group. (c,d) The proportion of T‐cell subpopulations in the skin‐draining lymph nodes of mice from each group. (e) Immunohistochemical staining of CD3 in the skin of mice from each group. (f) Immunohistochemical staining of Ki67 in the skin of mice from each group. Each group contains six mice and all experiments were repeated at least twice. Data show mean + SEM. P‐values were determined by one‐way analysis of variance with Bonferroni multiple‐comparisons t‐test. *P < 0·05, **P < 0·01, ***P < 0·001 and ****P < 0·0001

Article Snippet: Mouse recombinant IL‐33 (rIL‐33, 1 µg in 300 μl PBS; PeproTech, Rocky Hill, NJ) or PBS was injected subcutaneously daily into the back skin of the mice.

Techniques: Expressing, Immunohistochemical staining, Staining

Journal: Immunology

Article Title: Interleukin‐33 alleviates psoriatic inflammation by suppressing the T helper type 17 immune response

doi: 10.1111/imm.13203

Figure Lengend Snippet: Summary of interleukin‐33 (IL‐33) actions in psoriatic inflammation. IL‐17‐producing T cells [T helper type 17 (Th17), IL‐17‐producing γδT (γδT17) and IL‐17‐producing CD8+ T (Tc17) cells] produce and secrete large amounts of IL‐17A under the inflammatory microenvironment of psoriasis. Then IL‐17A acts on keratinocytes to promote the IL‐33 expression. On the one hand, the increased IL‐33 from keratinocytes or other cells in the dermis, such as fibroblasts, can promote the proliferation of keratinocytes to aggravate the psoriatic inflammation, but on the other hand, inhibit the differentiation and function of Th17 cells to remit the psoriatic inflammation

Article Snippet: Mouse recombinant IL‐33 (rIL‐33, 1 µg in 300 μl PBS; PeproTech, Rocky Hill, NJ) or PBS was injected subcutaneously daily into the back skin of the mice.

Techniques: Expressing