4 pge2  (Cayman Chemical)

Journal: EBioMedicine

Article Title: Pro-tumor Tfh2 cells induce detrimental IgG4 production and PGE 2 -dependent IgE inhibition in pancreatic cancer.

doi: 10.1016/j.ebiom.2023.104819

Figure Lengend Snippet: Fig. 7: In vitro-differentiated Tfh2 cells promote IgG4 and inhibit IgE secretion by B cells. (a) Experimental scheme: conditioned medium from co-cultures of tumor cells (BxPC3), CAFs, and primary monocytes was used to condition myeloid DCs (Sup-DC), which, in turn, were used to prime naïve CD4+ T cells. Untreated myeloid DCs (nt-DC) and TSLP-treated myeloid DCs (TSLP-DC) were used as controls. (b) Flow cytometry analysis of in vitro differentiated CXCR5+ T cells; contour plots and histograms show expression of PD-1 (high and low), ICOS, BCL6, CXCL13, and differentiation into Tfh1, Tfh2 and Tfh17 cells, based on CCR6 and CXCR3 (representative of n = 3). (c and d), Secretion of IL-21 (n = 3) (c), and other cytokines (d) by in vitro primed CD4+ T (predominantly Tfh2) cells activated as indicated. Results are expressed as fold increases in cytokine secretion relative to levels in nt-DC:T co-cultures (n = 4). (e) B cell help mediated by FACS-sorted CXCR5+ (red) and CXCR5−(blue) T cells in the absence/presence of an anti-IL-13Ra2 Ab (dashed columns) or an isotype control Ab (filled columns); representative of n = 2. CD40L + CpGB and CD40L + IL-13 + IL-21 were used as positive controls. (f) PGE2 secretion by in vitro-primed (TSLP-DC or Sup-DC) pre- dominant Tfh2 cells, expressed as fold increases over PGE2 levels produced by nt-DC:T co-cultures (n = 3). (g) IgG4 secreted by B cells cultured with CD40L alone or CD40L + IL-13 + IL-21 in the presence/absence of PGE2 (n = 8). (h) IgE secreted by B cells cultured as in (g) without/with the EP4 receptor antagonist (EP4RA). Significance was calculated using the paired Student’s t-test in (f) or one-way ANOVA followed by Newman–Keuls multiple comparison test in (c–e) and (g and h); *p < 0.05 and ***p < 0.001.

Article Snippet: For PGE2 functional assays, isolated B cells activated with M-CD40L, IL-13, and IL-21, as defined above, were treated ±1 μM PGE2 (14010, Cayman Chemical), and ±1 μM PGE2 receptor type 4 (EP4) antagonist (ONO-AE3-208, 3565, Tocris, Biotechne); culture supernatants were collected on day 10 for IgE and IgG4 quantification.

Techniques: In Vitro, Flow Cytometry, Expressing, Control, Produced, Cell Culture, Comparison

Journal: International Journal of Molecular Sciences

Article Title: A Combination of Celecoxib and Glucosamine Sulfate Has Anti-Inflammatory and Chondroprotective Effects: Results from an In Vitro Study on Human Osteoarthritic Chondrocytes

doi: 10.3390/ijms22168980

Figure Lengend Snippet: Celecoxib and GS reduce the expression of pro-inflammatory cytokines. Human osteoarthritic (OA) chondrocytes were incubated for 24 and 48 h with celecoxib (1.85 µM) and prescription-grade glucosamine sulfate (GS) (9 µM) (2 h of pre-treatment) in the presence of interleukin (IL)-1β (10 ng/mL). ( A – L ) Expression levels of cyclooxygenase ( COX)-2 , prostaglandin ( PG)E2 , IL-1β , IL-6 , and tumor necrosis factor (TNF)-α analyzed by quantitative real-time PCR. The gene expression was referenced to the ratio of the value of interest and the value of the basal condition, reported equal to 1. Data were represented as mean ± SD of triplicate values. * p < 0.05, ** p < 0.01, *** p < 0.001 versus basal condition. ° p < 0.05, °° p < 0.01 versus IL-1β .

Article Snippet: PGE2 production, the release of COX-2 , IL-1β , IL-6 , TNF-α , MMP-1 , MMP-3 , MMP-13 , and Col2a1 were detected using ELISA kits (Boster Biological Technology, CA, USA).

Techniques: Expressing, Incubation, Real-time Polymerase Chain Reaction, Gene Expression

Journal: International Journal of Molecular Sciences

Article Title: A Combination of Celecoxib and Glucosamine Sulfate Has Anti-Inflammatory and Chondroprotective Effects: Results from an In Vitro Study on Human Osteoarthritic Chondrocytes

doi: 10.3390/ijms22168980

Figure Lengend Snippet: Celecoxib and GS reduce the release of pro-inflammatory cytokines. Human osteoarthritic (OA) chondrocytes were incubated for 24 and 48 h with celecoxib (1.85 µM) and prescription-grade glucosamine sulfate (GS) (9 µM) (2 h of pre-treatment) in the presence of interleukin (IL)-1β (10 ng/mL). ( A – L ) Total amount of cyclooxygenase (COX)-2 , prostaglandin (PG)E2 , IL-1β , IL-6 , and tumor necrosis factor (TNF)-α , released in the supernatant, analyzed by ELISA assay. The total amount was referenced to the ratio of the value of interest and the value of the basal condition, reported equal to 1. Data were represented as mean ± SD of triplicate values. * p < 0.05, ** p < 0.01 versus basal condition. ° p < 0.05, °° p < 0.01 versus IL-1β .

Article Snippet: PGE2 production, the release of COX-2 , IL-1β , IL-6 , TNF-α , MMP-1 , MMP-3 , MMP-13 , and Col2a1 were detected using ELISA kits (Boster Biological Technology, CA, USA).

Techniques: Incubation, Enzyme-linked Immunosorbent Assay

Journal: International Journal of Molecular Sciences

Article Title: A Combination of Celecoxib and Glucosamine Sulfate Has Anti-Inflammatory and Chondroprotective Effects: Results from an In Vitro Study on Human Osteoarthritic Chondrocytes

doi: 10.3390/ijms22168980

Figure Lengend Snippet: BAY 11-7082 influences celecoxib and GS effects on pro-inflammatory cytokines. Human osteoarthritic (OA) chondrocytes were incubated for 24 h with celecoxib (1.85 µM) and prescription-grade glucosamine sulfate (GS) (9 µM) (2 h of pre-treatment) in the presence of interleukin (IL)-1β (10 ng/mL) and BAY 11-7082 1 μM (NF-κB inhibitor, 2 h of pre-treatment). ( A – E ) Expression levels of cyclooxygenase ( COX)-2 , prostaglandin (PG)E2 , IL-1β , IL-6 , and tumor necrosis factor (TNF)-α analyzed by quantitative real-time PCR. The gene expression was referenced to the ratio of the value of interest and the value of basal condition, reported equal to 1. Data were represented as mean ± SD of triplicate values. * p < 0.05, ** p < 0.01, *** p < 0.001 versus basal condition. ° p < 0.05, °° p < 0.01 versus IL-1β . # p < 0.05, ## p < 0.01 versus celecoxib or GS plus BAY.

Article Snippet: PGE2 production, the release of COX-2 , IL-1β , IL-6 , TNF-α , MMP-1 , MMP-3 , MMP-13 , and Col2a1 were detected using ELISA kits (Boster Biological Technology, CA, USA).

Techniques: Incubation, Expressing, Real-time Polymerase Chain Reaction, Gene Expression