anti st2 il 33r antibody  (R&D Systems)

Journal: bioRxiv

Article Title: IL-33 promotes transcriptional and metabolic adaptations of tissue-resident Th2 cells

doi: 10.1101/2025.07.09.663905

Figure Lengend Snippet: (A) A schematic of experimental design. CD4 T cells were isolated from Hpoly -infected mice and cultured with IL2 alone, IL2 and IL-33, or anti-CD3/CD28 under Th2 polarizing conditions for five days. (B) Representative flow plot of CD4 T cells immediately following isolation from secondary lymphoid organs of naïve or Hpoly -infected mice. (C) CD4 T cells from secondary lymphoid organs of Hpoly -infected mice were cultured under indicated conditions. Frequencies and numbers of GATA3+ST2+ cells were quantified. (D) Representative flow plot of CD4 T cells right after isolation from secondary lymphoid organs of Hpoly-infected 4get mice. (E) CD4 T cells from naïve or Hpoly-infected mice were cultured with IL2 and IL-33. (F) CD4 T cells from Hpoly-infected mice were cultured under the indicated conditions. Frequencies and numbers of GATA3+ST2+ cells were quantified after five days. (G) Abundance of amphiregulin in the culture supernatants from (F). (H) Splenocytes from naïve or Hpoly-infected mice cultured with IL2 alone or together with HES. Frequencies and numbers of GATA3+ cells was quantified. The data (B-H) are representative of at least two independent experiments. Symbols in the quantified data represent independent biological replicates. Data were analyzed by one-way ANOVA with Tukey’s post hoc test (B-G) or two-way ANOVA with Sidak post hoc test. * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001.

Article Snippet: Additionally, biotinylated antibodies were used for the staining ST2 (IL-33R) (MDB, clone DJ8) and amphiregulin (R&D Systems, #BAF989).

Techniques: Isolation, Infection, Cell Culture

Journal: bioRxiv

Article Title: IL-33 promotes transcriptional and metabolic adaptations of tissue-resident Th2 cells

doi: 10.1101/2025.07.09.663905

Figure Lengend Snippet: (A) Representative flow plots and quantification of ST2+GATA3 + in live CD45+TCRβ+CD4+FOXP3-T cells from mesenteric adipose tissue of Hpoly -infected mice or after five days of in vitro culture of CD4 T cells isolated from Hpoly-infected mice. Flow plots and quantification of ST2+IL4+(GFP) (B) or CD44+IL4+(GFP+) (C) cells cultured as above. (D) CD4 T cells from naïve or Hpoly -infected mice were cultured as in (A) and IL-33R+GATA3+ Th2 cells were quantified. (E) Representative flow plots and quantification of AREG+ cells within the IL4+(GFP+) population after 5hr restimulation with PMA/Iono in the presence of Brefeldin. The data (A-E) are representative of at least two independent experiments. Symbols in the quantified data represent independent biological replicates. Data were analyzed by one-way ANOVA with Tukey’s post hoc test (A-C, E) or two-way ANOVA with Sidak post hoc test (D).. * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001, *** p ≤ 0.0001.

Article Snippet: Additionally, biotinylated antibodies were used for the staining ST2 (IL-33R) (MDB, clone DJ8) and amphiregulin (R&D Systems, #BAF989).

Techniques: Infection, In Vitro, Isolation, Cell Culture

Journal: bioRxiv

Article Title: IL-33 promotes transcriptional and metabolic adaptations of tissue-resident Th2 cells

doi: 10.1101/2025.07.09.663905

Figure Lengend Snippet: (A) CD44+IL4+ (GFP)+ early Th2 cells, CD44+IL4-(GPF-) activated cells, and naïve CD44-GFP-cells were FACS-isolated from 4get mice infected with Hpoly and cultured with IL2 and IL-33 for five days. Frequencies of GFP+ and GFP+ST2+ cells (B) as well as ST2 gMFI were quantified (C). The data (A-C) are representative of at least two independent experiments. Symbols in the quantified data represent independent biological replicates. Data were analyzed by one-way ANOVA with Tukey’s post hoc test (B) or paired T-test (C). * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001.

Article Snippet: Additionally, biotinylated antibodies were used for the staining ST2 (IL-33R) (MDB, clone DJ8) and amphiregulin (R&D Systems, #BAF989).

Techniques: Isolation, Infection, Cell Culture

Journal: bioRxiv

Article Title: IL-33 promotes transcriptional and metabolic adaptations of tissue-resident Th2 cells

doi: 10.1101/2025.07.09.663905

Figure Lengend Snippet: RNAseq was performed on CD44+IL4+(GFP+) Th2 cells isolated from spleens and lymph nodes of Hpoly -infected mice or after five-day culture under indicated conditions. (A) Principal component analysis of RNAseq data. (B) Heatmap of hierarchical clustering of differentially expressed genes. (C) Top Hallmark pathways from GSEA for IL-33 v IL2 comparison. (D) Flow plots and quantification of pS6 expression in CD44+IL4(GFP+) Th2 cells. (E) CD4 T cells were isolated from Hpoly -infected mice and cultured with IL2 and IL-33 in the presence of rapamycin. Frequencies and numbers of GATA3+ST2+ Th2 cells were quantified. (F) Levels of indicated cytokines after overnight culture in fresh media containing rapamycin. The data (D, E, F) are representative of at least two independent experiments. Symbols in the quantified data represent independent biological replicates. Data were analyzed by one-way ANOVA with Tukey’s post hoc test (D, E) or paired T-test (F). * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001.

Article Snippet: Additionally, biotinylated antibodies were used for the staining ST2 (IL-33R) (MDB, clone DJ8) and amphiregulin (R&D Systems, #BAF989).

Techniques: Isolation, Infection, Comparison, Expressing, Cell Culture

Journal: bioRxiv

Article Title: IL-33 promotes transcriptional and metabolic adaptations of tissue-resident Th2 cells

doi: 10.1101/2025.07.09.663905

Figure Lengend Snippet: (A) Top Hallmark pathways from GSEA for IL-33 v IL2 comparison. (B) CD4 T cells were isolated from Hpoly -infected mice and cultured anti-CD3/CD28 in the presence of rapamycin. The mean fluorescence intensity of GATA3 and cell numbers were quantified. (C) Levels of indicated cytokines after overnight culture of anti-CD3/CD28-stimulated cells in fresh media containing rapamycin. (D) Quantification of results from SCENITH assay in GATA3+ Th2 cells. (E) Representative flow plot showing the expression of TSLPR and ST2 after five days of in vitro culture of CD4 T cells were isolated from Hpoly -infected mice. (F) Representative flow plot and quantification of mitotracker green and mitotracker deep red staining after five days of culture. The data (B-F) are representative of at least two independent experiments. Symbols in the quantified data represent independent biological replicates. Data were analyzed by one-way ANOVA with Tukey’s post hoc test (B, E, F), or paired T-test (C). * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001.

Article Snippet: Additionally, biotinylated antibodies were used for the staining ST2 (IL-33R) (MDB, clone DJ8) and amphiregulin (R&D Systems, #BAF989).

Techniques: Comparison, Isolation, Infection, Cell Culture, Fluorescence, Expressing, In Vitro, Staining

Journal: bioRxiv

Article Title: IL-33 promotes transcriptional and metabolic adaptations of tissue-resident Th2 cells

doi: 10.1101/2025.07.09.663905

Figure Lengend Snippet: (A) Heatmap representing the abundance of the top 100 detected metabolites. (B) CD4 T cells were isolated from Hpoly -infected mice and cultured with IL2 and IL-33 for five days. The intracellular abundance of indicated amino acids in naïve and CD4+CD44+ activated cells was determined via LC/MS. (C) Heatmap depicting the expression of arginine transporters genes related in RNAseq data from . (D) CD4 T cells were isolated from Hpoly -infected mice and cultured with IL2 and IL-33 for five days in amino acid drop-out media. Frequencies and numbers of GATA3+ST2+ Th2 cells were quantified. (E) Representative histograms and quantification of CTV dilution of CD4 T cells cultured as in D. (F) After five days of culture with IL2 and IL-33 in control or arginine-depleted media, cells were restimulated with PMA/Iono and amphiregulin was quantified. (G) Representative histograms and quantification of ps6 levels in pS6-positive cells at day 1 of culture. The data (D-G) are representative of at least two independent experiments. Data were analyzed by one-way ANOVA with Tukey’s post hoc test (D, E), paired T-test (F), or two-way ANOVA with Sidak post hoc test. * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001.

Article Snippet: Additionally, biotinylated antibodies were used for the staining ST2 (IL-33R) (MDB, clone DJ8) and amphiregulin (R&D Systems, #BAF989).

Techniques: Isolation, Infection, Cell Culture, Liquid Chromatography with Mass Spectroscopy, Expressing, Control

Journal: bioRxiv

Article Title: IL-33 promotes transcriptional and metabolic adaptations of tissue-resident Th2 cells

doi: 10.1101/2025.07.09.663905

Figure Lengend Snippet: (A) Expresssion of Th2 cell related genes in scRNAseq dataset of T cells isolated from mAT of Hpoly -infected mice identifying cluster of GATA3+ST2+ Th2 cells (circled). CD4 T cells from Hpoly-infected mice were stimulated with anti-CD3/CD28 and treated with DFMO (B) or cultured in arginine-depleted media (C). Mean fluorescence intestity of GATA3 and cells numbers were quantified. The data (B,C) are representative of at least two independent experiments. Symbols in the quantified data represent independent biological replicates. Data were analyzed by one-way ANOVA with Tukey’s post hoc test (B, C). * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001.

Article Snippet: Additionally, biotinylated antibodies were used for the staining ST2 (IL-33R) (MDB, clone DJ8) and amphiregulin (R&D Systems, #BAF989).

Techniques: Isolation, Infection, Cell Culture, Fluorescence