hek blue il 33 cells  (InvivoGen)

Journal: Frontiers in Immunology

Article Title: Microbial sensing through the non-canonical inflammasome modulates airway type 2 immunity

doi: 10.3389/fimmu.2026.1784561

Figure Lengend Snippet: Non-canonical inflammasome-dependent pyroptosis and IL-33 secretion in airway epithelial cells. (A) Western blot from lysates of primary human airway epithelial cells cultured for 16 hours following electroporation of LPS or dA:dT (or sham). (B) Western blot of GSDMD and GAPDH from lysates of BEAS-2B cells treated with IC LPS and dA:dT for 3 hours or nigericin for 1 hour. (C) Glucose 6-phosphade dehydrogenase (G6PD) activity measured in media from BEAS-2B cells treated with IC LPS, dA:dT, or nigericin for 16 hours. (D) Western blot of IL-33 in concentrated supernatant from IL-33 OE BEAS-2B or GSDMD crispr BEAS-2B cells. (E) IL-33 activity measured by secreted embryonic alkaline phosphatase from HEK-Blue-IL33 cells treated with media from IL-33 OE BEAS-2B stimulated for 16 hours with IC LPS. (F) Gene expression for mitogen activated protein kinase (MAPK) target genes in BEAS-2B cells treated with IC LPS or dA:dT and nigericin for 4 hours. (G) IL33 gene expression in BEAS-2B cells treated with IC LPS or MAPK inhibitor (MAPKi, trametinib) for 4 hours. (H) Western blot of GSDMD and GAPDH from cellular lysates from CRISPR- BEAS-2B cell lines treated with IC LPS for 3 hours. (I) Propidium iodide (PI) cell uptake over time in CRISPR- BEAS-2B cell lines with indicated knockdowns. (J) IL-33 activity measured by secreted embryonic alkaline phosphatase from HEK-Blue-IL33 cells treated with media from IL33 OE CRISPR- BEAS-2B stimulated for 16 hours with IC LPS. (K) Gene expression for MAPK target genes and IL33 from CRISPR-BEAS-2B stimulated for 4 hours with IC LPS. Data points reflect biological replicates. Error bars represent SEM. p values represent ordinary ANOVA with Sidak correction (C, J) or Tukey correction (E) .

Article Snippet: HEK-Blue IL-33 Cells (Invivogen cat hkb-hil33) were cultured according to manufacturer’s instructions.

Techniques: Western Blot, Cell Culture, Electroporation, Activity Assay, CRISPR, Gene Expression

Journal: Frontiers in Immunology

Article Title: Microbial sensing through the non-canonical inflammasome modulates airway type 2 immunity

doi: 10.3389/fimmu.2026.1784561

Figure Lengend Snippet: Caspase4-dependent epithelial pyroptosis and IL-33 secretion are modulated by papain via intracellular calcium. (A) Propidium iodide (PI) uptake at 6 hours in cells treated with IC LPS and papain. (B) GFP ELISA on conditioned media from IL-33 OE BEAS-2B cells treated with IC LPS and papain or heat inactivated (HI) papain for 16 hours. (C) IL-33 activity measured by secreted embryonic alkaline phosphatase from HEK-Blue-IL33 cells treated with media from IL-33OE BEAS-2B cells treated with IC LPS, papain and HI papain for 16 hours. (D) Intracellular calcium measured in BEAS-2B cells following thapsigargin, papain, or papain and BTP2. (E) PI uptake at 6 hours in BEAS-2B cells treated with IC LPS, papain and BTP2. (F) GFP ELISA on conditioned media from IL-33 OE BEAS-2B cells treated with IC LPS, papain and BTP2 for 16 hours. (G) PI uptake in BEAS-2B cells lines treated with IC LPS and calcium chloride. (H) GFP ELISA on conditioned media from IL33OE BEAS-2B cells treated with IC LPS and calcium chloride 16 hours. (I) IL-33 activity measured via HEK-Blue-IL33 cells in media from IL-33OE BEAS-2B cells treated with IC LPS and calcium chloride for 16 hours. (J) Western blot of GSDMD and GAPDH from cellular lysates from BEAS-2B cells treated with IC LPS and calcium chloride for 3 hours. P value represents two-way ANOVA with Tukey correction. (K) Western blot of BAL showing IL-33 protein in wildtype mice treated with 3 days of intratracheal PBS or papain. ( L ) Eosinophil and ILC2 counts from lungs of wildtype or Casp4 -/- mice treated with 3 days of intratracheal PBS or papain. Data points reflect biological replicates. Error bars indicate SEM. P values indicated for ordinary one-way ANOVA with Tukey correction.

Article Snippet: HEK-Blue IL-33 Cells (Invivogen cat hkb-hil33) were cultured according to manufacturer’s instructions.

Techniques: Enzyme-linked Immunosorbent Assay, Activity Assay, Western Blot

Journal: Frontiers in Immunology

Article Title: Microbial sensing through the non-canonical inflammasome modulates airway type 2 immunity

doi: 10.3389/fimmu.2026.1784561

Figure Lengend Snippet: LPS from different gram-negative bacteria differ in ability to activate non-canonical inflammasome and IL-33 secretion in airway epithelia. (A) Western blot of GSDMD and GAPDH from cellular lysates from BEAS-2B cells treated for 3 hrs with purified LPS from three different bacterial species: Escherichia coli 55:B5 ( E.coli ), Pseudomonas aeruginosa 10.22 ( Ps.A ) and Klebsiella pneumonia ( Kleb ). (B) Western blot of IL-33 in concentrated supernatant from BEAS-2B cells treated for 16 hrs with IC LPS from different bacterial species. (C) Propidium iodide (PI) cell uptake over time in BEAS-2B cells lines treated with IC LPS from different bacterial species. (D) Propidium iodide (PI) cell uptake at 6 hours in CRISPR-BEAS-2B cells lines treated with IC LPS from different bacterial species. (E) MAPK and (F) IL33 gene expression from BEAS-2B cells treated for 4 hours with IC LPS from different bacterial species. Data points reflect biological replicates. Error bars indicate SEM. P values indicated for ordinary one-way ANOVA with Tukey correction.

Article Snippet: HEK-Blue IL-33 Cells (Invivogen cat hkb-hil33) were cultured according to manufacturer’s instructions.

Techniques: Bacteria, Western Blot, Purification, CRISPR, Gene Expression

Journal: mAbs

Article Title: Structures of clinical antibodies bound to IL-33 uncover two distinct epitopes underlying differential efficacy

doi: 10.1080/19420862.2026.2639673

Figure Lengend Snippet: Characterization of the three anti-IL-33 antibodies. (A) Sequence alignment of etokimab, itepekimab, and tozorakimab. CDRs and FRs were defined according to the IMGT system. (B) SPR analysis measuring the binding affinity of the three antibodies and ST2 to IL-33. Serially diluted IL-33 (starting concentration 11.11 nM) was injected over immobilized ligands for a 180 s association and 300 s dissociation. (C) Inhibition of IL-33-induced signaling by the antibodies. HEK-Blue IL-33 reporter cells were stimulated with IL-33 at 2 ng/mL (triangles) or 30 ng/mL (circles) in the presence of a series of each antibody. After 22 hours, SEAP activity in the supernatant was quantified by measuring absorbance at 620 nm. Itepekimab, etokimab, and tozorakimab are shown in blue, green, and purple, respectively. Data points represent mean ± SD ( n = 3). Dose–response curves were fitted, and IC 50 /IC 90 values were calculated using GraphPad Prism which were summarized in the right panel table. (D) Blocking of IL-33–induced signaling on human peripheral blood mononuclear cell (PBMCs). Antibody and 1 nM IL-33 were added to human PBMCs for 19 hours. IFN-γ release was measured by means of ELISA. All data are represented as means ± SDs.

Article Snippet: HEK-BlueTM IL-33 cells (InvivoGen, catalog no. hkb-hil33) were utilized for bioactivity assessment of IL-33 through monitoring NF-κB and AP-1 pathway activation.

Techniques: Sequencing, Binding Assay, Concentration Assay, Injection, Inhibition, Activity Assay, Blocking Assay, Enzyme-linked Immunosorbent Assay

Journal: mAbs

Article Title: Structures of clinical antibodies bound to IL-33 uncover two distinct epitopes underlying differential efficacy

doi: 10.1080/19420862.2026.2639673

Figure Lengend Snippet: Structural overview of antibody – IL-33 complexes. (A) Structures of IL-33 (medium orchid) in complex with etokimab Fab (salmon), itepekimab Fab (olive drab), and tozorakimab Fab (dark turquoise), shown from left to right. The rightmost panel shows a superposition of all three Fab – IL-33 complexes, illustrating their relative positions and orientations. (B) Structure of the IL-33–ST2 complex (dark slate blue; PDB: 4KC3). The interface of D1/D2 domains and IL-33 named Site 1, interface of D3 domain and IL-33 named Site 2. (C) Structural alignment of the three Fab – IL-33 complexes with the ST2–IL-33 complex, showing the spatial relationship between each antibody and ST2. Colors are consistent with panels (A) and (B).

Article Snippet: HEK-BlueTM IL-33 cells (InvivoGen, catalog no. hkb-hil33) were utilized for bioactivity assessment of IL-33 through monitoring NF-κB and AP-1 pathway activation.

Techniques:

Journal: mAbs

Article Title: Structures of clinical antibodies bound to IL-33 uncover two distinct epitopes underlying differential efficacy

doi: 10.1080/19420862.2026.2639673

Figure Lengend Snippet: Interactions of etokimab and itepekimab with IL-33. (A, B) Epitopes on IL-33 (medium orchid) and paratopes on (A) etokimab (heavy chain, salmon; light chain, orange) and (B) itepekimab (heavy chain, olive drab; light chain, coral) are colored in dark gray, binding residues within are highlighted. (C, D) Detailed interaction networks for (C) Etokimab – IL-33 and (D) Itepekimab – IL-33 complexes. Hydrogen bonds and salt bridges are indicated by black dashed and solid lines, respectively; a red solid line denotes an interaction involving both. Interaction residues within are highlighted in related color. (E) Structural footprint of etokimab (salmon), itepekimab (olive drab), tozorakimab (dark turquoise), and ST2 (dark slate blue) on IL-33 (dark gray). Shared binding residues are listed in the lower panel.

Article Snippet: HEK-BlueTM IL-33 cells (InvivoGen, catalog no. hkb-hil33) were utilized for bioactivity assessment of IL-33 through monitoring NF-κB and AP-1 pathway activation.

Techniques: Binding Assay

Journal: mAbs

Article Title: Structures of clinical antibodies bound to IL-33 uncover two distinct epitopes underlying differential efficacy

doi: 10.1080/19420862.2026.2639673

Figure Lengend Snippet: Molecular basis of tozorakimab recognition of IL-33. (A) Binding interface between tozorakimab and IL-33. The epitope on IL-33 is shown in dark gray, with key contacting residues highlighted. The paratope is composed of the heavy chain (dark turquoise) and light chain (yellow-green). (B) Structural footprint comparison between tozorakimab and ST2 on IL-33. The tozorakimab footprint is shown in dark turquoise, while the ST2 Site 1 interface is colored dark slate blue. Shared binding residues are colored and highlighted according to their chain origin in tozorakimab. (C) Detailed molecular interactions in the tozorakimab – IL-33 complex. Hydrogen bonds and salt bridges are indicated by black dashed and solid lines, respectively; a red solid line denotes an interaction involving both. Relevant residues are colored consistently with panels A and B. (D) Electrostatic surface potential analysis of IL-33 (left) and tozorakimab (right). Surfaces are colored from red (negative charge) to blue (positive charge). The binding footprints of tozorakimab (dark turquoise) and IL-33 (medium orchid) are outlined. Dashed circles indicate complementary charged patches that facilitate binding.

Article Snippet: HEK-BlueTM IL-33 cells (InvivoGen, catalog no. hkb-hil33) were utilized for bioactivity assessment of IL-33 through monitoring NF-κB and AP-1 pathway activation.

Techniques: Binding Assay, Comparison

Journal: mAbs

Article Title: Structures of clinical antibodies bound to IL-33 uncover two distinct epitopes underlying differential efficacy

doi: 10.1080/19420862.2026.2639673

Figure Lengend Snippet: Characterization of tozorakimab and related mutants. (A) Binding affinity of tozorakimab and mutants to IL33 was assessed by indirect ELISA. Wells were coated with IL33 (2 μg/mL). An anti-human IgG-HRP antibody (promega, cat. no. W4031 ) was used for detection. (B) Inhibition of IL-33-induced signaling by the tozorakimab and mutants. HEK-Blue IL-33 reporter cells were stimulated with IL-33 at 0.5 nM in the presence of a series of each antibody. After 22 hours, SEAP activity in the supernatant was quantified by measuring absorbance at 620 nm. Data points represent mean ± SD ( n = 3). Dose – response curves were fitted and EC 50 or IC 50 values were calculated using GraphPad prism ( n = 3); results are summarized in the right panel table.

Article Snippet: HEK-BlueTM IL-33 cells (InvivoGen, catalog no. hkb-hil33) were utilized for bioactivity assessment of IL-33 through monitoring NF-κB and AP-1 pathway activation.

Techniques: Binding Assay, Indirect ELISA, Inhibition, Activity Assay

Journal: mAbs

Article Title: Structures of clinical antibodies bound to IL-33 uncover two distinct epitopes underlying differential efficacy

doi: 10.1080/19420862.2026.2639673

Figure Lengend Snippet: SPR competition assays of three antibodies and ST2. IL-33 was immobilized on a CM5 sensor chip. Injections were performed in two steps: first, protein 1 was injected for 150 s, followed by a mixture of protein 1 and protein 2 for 150 s. From left to right: competition profiles for itepekimab + ST2, etokimab + ST2, and tozorakimab + ST2, ST2 = 200 nM，Abs = 100 nM.

Article Snippet: HEK-BlueTM IL-33 cells (InvivoGen, catalog no. hkb-hil33) were utilized for bioactivity assessment of IL-33 through monitoring NF-κB and AP-1 pathway activation.

Techniques: Injection