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tlr8 antagonist cu cpt9a  (Tocris)
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Tocris tlr8 antagonist cu cpt9a
Figure 4. Dual neutralizing TLR2 mAb and CQ application in vitro abrogates Streptococcus pneumoniae–induced cytokine release by murine and human immune cells. (A and B) Murine BMDMs (blue boxes) and (C–F) human PBMCs (green boxes) were cultured and challenged with 1 × 106 CFU/mL viable S. pneumoniae (SP, except for controls, which received THY solution only). Four hours postinfection (p.i.), cells were treated with ceftriaxone. Two hours earlier (2 h), simultaneously (4 h), or 2 hours later (6 h p.i.), anti-TLR2 mAb (T2.5) and chloroquine (CQ) were added to cell cultures. In human PBMC cultures, the <t>TLR8</t> inhibitor <t>Cu-Cpt9a</t> and dexamethasone (DEXA) were applied additionally where indicated. Cell culture supernatants were sampled 24 hours p.i., aliquoted, and stored until ELISA analysis. Supernatants were analyzed for IL-6 (A–D), IL-1β (E), and TNF (F). The experiments with BMDMs were conducted at least 2 times in triplicate, while those involving human PBMCs were performed at least 2 times in quadruplicate (for detailed information see Supporting Data Values file). The data for the placebo and T2.5 plus CQ (4 h) groups in A and B (as indicated by blue boxes with gray diagonal stripes in B) largely overlap. Data are depicted as median (line in box), 25%–75% percentile range (bounds of the box), and minimum and maximum (whiskers). Statis- tical test was 1-way ANOVA and subsequent Tukey’s post hoc test. ****P < 0.0001; ***P < 0.001; **P < 0.01; *P < 0.05. NS, not significant.
Tlr8 Antagonist Cu Cpt9a, supplied by Tocris, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Figure 4. Dual neutralizing TLR2 mAb and CQ application in vitro abrogates Streptococcus pneumoniae–induced cytokine release by murine and human immune cells. (A and B) Murine BMDMs (blue boxes) and (C–F) human PBMCs (green boxes) were cultured and challenged with 1 × 106 CFU/mL viable S. pneumoniae (SP, except for controls, which received THY solution only). Four hours postinfection (p.i.), cells were treated with ceftriaxone. Two hours earlier (2 h), simultaneously (4 h), or 2 hours later (6 h p.i.), anti-TLR2 mAb (T2.5) and chloroquine (CQ) were added to cell cultures. In human PBMC cultures, the TLR8 inhibitor Cu-Cpt9a and dexamethasone (DEXA) were applied additionally where indicated. Cell culture supernatants were sampled 24 hours p.i., aliquoted, and stored until ELISA analysis. Supernatants were analyzed for IL-6 (A–D), IL-1β (E), and TNF (F). The experiments with BMDMs were conducted at least 2 times in triplicate, while those involving human PBMCs were performed at least 2 times in quadruplicate (for detailed information see Supporting Data Values file). The data for the placebo and T2.5 plus CQ (4 h) groups in A and B (as indicated by blue boxes with gray diagonal stripes in B) largely overlap. Data are depicted as median (line in box), 25%–75% percentile range (bounds of the box), and minimum and maximum (whiskers). Statis- tical test was 1-way ANOVA and subsequent Tukey’s post hoc test. ****P < 0.0001; ***P < 0.001; **P < 0.01; *P < 0.05. NS, not significant.

Journal: JCI insight

Article Title: Combining antibiotic with anti-TLR2/TLR13 therapy prevents brain pathology in pneumococcal meningitis.

doi: 10.1172/jci.insight.165737

Figure Lengend Snippet: Figure 4. Dual neutralizing TLR2 mAb and CQ application in vitro abrogates Streptococcus pneumoniae–induced cytokine release by murine and human immune cells. (A and B) Murine BMDMs (blue boxes) and (C–F) human PBMCs (green boxes) were cultured and challenged with 1 × 106 CFU/mL viable S. pneumoniae (SP, except for controls, which received THY solution only). Four hours postinfection (p.i.), cells were treated with ceftriaxone. Two hours earlier (2 h), simultaneously (4 h), or 2 hours later (6 h p.i.), anti-TLR2 mAb (T2.5) and chloroquine (CQ) were added to cell cultures. In human PBMC cultures, the TLR8 inhibitor Cu-Cpt9a and dexamethasone (DEXA) were applied additionally where indicated. Cell culture supernatants were sampled 24 hours p.i., aliquoted, and stored until ELISA analysis. Supernatants were analyzed for IL-6 (A–D), IL-1β (E), and TNF (F). The experiments with BMDMs were conducted at least 2 times in triplicate, while those involving human PBMCs were performed at least 2 times in quadruplicate (for detailed information see Supporting Data Values file). The data for the placebo and T2.5 plus CQ (4 h) groups in A and B (as indicated by blue boxes with gray diagonal stripes in B) largely overlap. Data are depicted as median (line in box), 25%–75% percentile range (bounds of the box), and minimum and maximum (whiskers). Statis- tical test was 1-way ANOVA and subsequent Tukey’s post hoc test. ****P < 0.0001; ***P < 0.001; **P < 0.01; *P < 0.05. NS, not significant.

Article Snippet: PBMCs and BMDMs were challenged in identical manners except for the treatment with dexamethasone (5 μg/ mL) and the TLR8 antagonist Cu-Cpt9a (10 μg/mL; Tocris) (66).

Techniques: In Vitro, Cell Culture, Enzyme-linked Immunosorbent Assay