j113863 ccr1 antagonist tocris  (Tocris)

Journal: Frontiers in Molecular Biosciences

Article Title: Multitargeted biological actions of polydatin in preventing pseudogout acute attack

doi: 10.3389/fmolb.2025.1553912

Figure Lengend Snippet: Effect of CCR1 inhibition on PD treatment. Monocytes were pretreated with PD (100 µM) for 2 h and then stimulated for 24 h with CPP (0.025 mg/mL). Where indicated, J113863 (10 µM) was added 30 min before the PD treatment. (A) IL-1β, (B) IL-18, (C) IL-6, (D) TNFα, (E) IL-8, (F) VEGF levels in supernatants were quantified by ELISA. Data are expressed as mean of four independent experiments ±SD. p calculated according to the One Way Anova, Bonferroni’s multiple comparisons test:*p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. p calculated according to t-test, PD + CPP vs J + PD + CPP: °°p < 0.01. CPP, calcium pyrophosphate crystals; PD, polydatin.

Article Snippet: Where indicated, polydatin (100 µM) was added 2 h before the stimulation with crystals and anakinra (0.1 µg/mL), J113863 (Medchemexpress, Monmouth Junction, USA, 10 µM) and EX527 (Selleckchem, Cologne, 10 µM) were added 30 min before the use of polydatin.

Techniques: Inhibition, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Molecular Biosciences

Article Title: Multitargeted biological actions of polydatin in preventing pseudogout acute attack

doi: 10.3389/fmolb.2025.1553912

Figure Lengend Snippet: Migration of PBMCs exposed to SFs from patients with CPP induced arthritis in the presence or absence of PD (100–200 µM) or J113866 (10 µM) for 1.30 h. Medium RMPI supplemented with 5% plasma was used as a positive control (C+), while medium RPMI supplemented with 1% FBS was used as a negative control (C-). Left panel : Representative image of migrated cells on the bottom of a filter membrane of a modified 48-well Boyden chamber. Right panel : Effect of 100–200 μM PD on PBMCs migration induced by (A) 5% plasma (n = 3) (B) inflammatory SFs from CIA patients (n = 4). (C) Effect of 10 μM J113863 on PBMCs migration induced by inflammatory SFs from CIA patients (n = 4). Cell migration is shown as optical density values (O.D). Each sample was tested in sextuplicate. Data are expressed as mean of three to four independent experiments ±SD. p calculated according to the One Way Anova, Bonferroni’s multiple comparisons test: *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. CPP, calcium pyrophosphate crystals; PD, polydatin; SF, synovial fluid; PBMCs, peripheral blood mononuclear cells.

Article Snippet: Where indicated, polydatin (100 µM) was added 2 h before the stimulation with crystals and anakinra (0.1 µg/mL), J113863 (Medchemexpress, Monmouth Junction, USA, 10 µM) and EX527 (Selleckchem, Cologne, 10 µM) were added 30 min before the use of polydatin.

Techniques: Migration, Positive Control, Negative Control, Membrane, Modification

Journal: Biomedicines

Article Title: Analysing the Combined Effects of Radiotherapy and Chemokine Receptor 5 Antagonism: Complementary Approaches to Promote T Cell Function and Migration in Oesophageal Adenocarcinoma

doi: 10.3390/biomedicines12040819

Figure Lengend Snippet: Expanded circulating T cells from OAC patients expressed higher levels of CCR5, which was further upregulated on the surface of CD8 + T cells by clinically relevant doses of irradiation. ( a ) PBMCs isolated from treatment−naïve OAC donors ( n = 7) and age—matched HDs ( n = 6) were activated with plate−bound anti−CD3 and anti−CD28 agonists for 72 h, receiving 2 × 1.8 Gy fractions of irradiation (irradiated, IR) on day 1 and day 2, 24 h apart, or were non−irradiated, NIR. The frequencies of T cells in HDs and CDs expressing CCR1 ( b ), CCR5 ( c , d ), and CX 3 CR1 ( e ) were assessed by flow cytometry. The effect of IR on CCR1, CCR5, and CX 3 CR1 expression on T cells from HDs ( f ) and CDs ( g ) was also assessed by flow cytometry and depicted in heat maps shown (% cells expressing). The effect of IR on CCR5 expression on T cells from CDs is displayed in ( h , i ). All analyses were conducted on viable T cells using a zombie dye to exclude dead cells, and fluorescence minus-one controls (FMO) were used for gating analysis. Mann−Whitney tests were used to compare between HD and CD groups. Wilcoxon signed-rank tests were used to compare the effect of NIR with IR in the same donors. * p < 0.05, ** p < 0.01.

Article Snippet: For experiments that included treatment with antagonists, in the last 24 h of the T cell activation process, PBMCs were treated with 1 nM CCR1 antagonist J113863 (Axon MedChem, Reston, VA, USA), 80 μM of CCR5 antagonist Maraviroc (Axon MedChem, Reston, VA, USA), 245 nM CX 3 CR1 antagonist AZD8798 (Axon MedChem, Reston, VA, USA), or vehicle control (0.01% DMSO).

Techniques: Irradiation, Isolation, Expressing, Flow Cytometry, Fluorescence, MANN-WHITNEY

Journal: Biomedicines

Article Title: Analysing the Combined Effects of Radiotherapy and Chemokine Receptor 5 Antagonism: Complementary Approaches to Promote T Cell Function and Migration in Oesophageal Adenocarcinoma

doi: 10.3390/biomedicines12040819

Figure Lengend Snippet: CCR5 antagonism significantly increased IFN-γ production by OAC patient−derived CD4 + T helper cells. ( a ) PBMCs isolated from treatment-naïve OAC donors ( n = 5) and age−matched non-cancer donors ( n = 6) were activated with anti-CD3 and anti-CD28 agonists for 72 h and treated with J113863 (CCR1 antagonist) and RANTES, AZD8987 (CX 3 CR1 antagonist) and fractalkine or Maraviroc (CCR5 antagonist) and MIP-1α. The PBMCs also received 2 × 1.8 Gy fractions of irradiation on day 1 and day 2, 24 h apart, or were non-irradiated (NIR). The frequency of cells producing IFN-γ and CD107a was assessed by intracellular and extracellular flow cytometry, respectively. ( b – d ) heat maps depicting the effect of activating or antagonising the CCR1−RANTES, CCR5-MIP-1α, and CX 3 CR1−fractalkine pathways on the percentage of CD4 + and CD8 + T cells producing IFN-γ, respectively, relative to the untreated control. ( e ) Graphical display showing the effect of CCR5 antagonism on the production of IFN-γ by T cells, with representative dot plots shown in ( f ). ( g ) The effect of activating or antagonizing CCR1-RANTES, CCR5-MIP-1α, and CX 3 CR1-fractalkine pathway on CD107a degranulation by CD8 + T cells. All analyses were conducted on viable T cells using a zombie dye to exclude dead cells, and FMO controls were used for gating analysis. Paired parametric t -test was used to compare between two groups * p < 0.05.

Article Snippet: For experiments that included treatment with antagonists, in the last 24 h of the T cell activation process, PBMCs were treated with 1 nM CCR1 antagonist J113863 (Axon MedChem, Reston, VA, USA), 80 μM of CCR5 antagonist Maraviroc (Axon MedChem, Reston, VA, USA), 245 nM CX 3 CR1 antagonist AZD8798 (Axon MedChem, Reston, VA, USA), or vehicle control (0.01% DMSO).

Techniques: Derivative Assay, Isolation, Irradiation, Flow Cytometry, Control