Journal: Journal of Inflammation (London, England)

Article Title: Salivary histatin 3 inhibits heat shock cognate protein 70-mediated inflammatory cytokine production through toll-like receptors in human gingival fibroblasts

doi: 10.1186/1476-9255-11-4

Figure Lengend Snippet: The effect of HSC70 on NF-κB-dependent activation through TLR2 and TLR4. (A) Luciferase assays for NF-κB-dependent activation by HSC70 through TLR4. 293-TLR4/MD2-CD14 cells were transfected with an NF-κB-dependent luciferase reporter plasmid (pIgκB-Luc). One day after transfection, cells were stimulated with heated and unheated E. coli LPS, BSA, HSP70, or HSC70 for 6 h. Cell lysates prepared from the stimulated cells were analyzed by luciferase assays. The values are shown as fold induction of the standardized luciferase activity over the unstimulated control (none). (B) Luciferase assays for NF-κB-dependent activation by the ATPase fragment of HSC70 in 293-TLR4/MD2-CD14 cells. The procedures were as in (A) , except that the cells were stimulated with the heated or unheated HSC70 ATPase fragment. (C) Luciferase assays for NF-κB-dependent activation by HSC70 through TLR2. The procedures were as in (A) , except that 293-TLR2/CD14 cells and P. gingivalis LPS were used. (D) Luciferase assays for NF-κB-dependent activation by the HSC70 ATPase fragment in 293-TLR2/CD14 cells. The procedures were as in (A and B). Bars represent the means and range of duplicate samples.

Article Snippet: ; Nippon Kayaku); mouse monoclonal anti-TLR4 (HTA125) and anti-TLR2 (TL2.1) antibodies (Hycult Biotechnology); mouse monoclonal anti-CD14 (MY4) antibody (Beckman Coulter); rabbit polyclonal anti-p44/42, anti-p38, anti-phosphorylated p38, anti-phosphorylated IκB-α, mouse monoclonal anti-phosphorylated p44/42 antibodies (Cell Signaling); mouse monoclonal anti-JUN-N-terminal protein kinase (JNK) (D-2), anti-phosphorylated JNK (G-7), anti-IκB-α (H-4) antibodies (Santa Cruz Biotechnology); and mouse monoclonal anti-β-actin antibody (Abcam).

Techniques: Activation Assay, Luciferase, Transfection, Plasmid Preparation, Activity Assay

Journal: Journal of Inflammation (London, England)

Article Title: Salivary histatin 3 inhibits heat shock cognate protein 70-mediated inflammatory cytokine production through toll-like receptors in human gingival fibroblasts

doi: 10.1186/1476-9255-11-4

Figure Lengend Snippet: The effect of histatin 3 on NF-κB-dependent activation by HSC70 through TLR2 and TLR4. (A, B) Luciferase assays for NF-κB-dependent activation by HSC70 through TLR2 and TLR4 in the presence of histatin 3. 293-TLR4/MD2-CD14 (A) and 293-TLR2/CD14 (B) cells were transfected with pIgκB-Luc, and were stimulated with mixtures of the indicated peptides and LPS, HSP70, or HSC70 for 6 h. Cell lysates prepared from the stimulated cells were analyzed by luciferase assays. The values are shown as fold induction of the standardized luciferase activity over the unstimulated control (none). **, P < 0.01; ***, P < 0.001 versus stimulation with HSC70 alone. cont. pep., control peptide. (C, D) Luciferase assays for NF-κB-dependent activation by HSC70 through TLR2 and TLR4 in the presence of histatin family members. The procedures were same as in (A and B), except that stimulations were with mixtures of HSC70 and histatins 3, 4, or 5. Bars represent the means and range of duplicate samples. *, P < 0.05; **, P < 0.01 versus stimulation with HSC70 alone. †, P < 0.05; ††, P < 0.01.

Article Snippet: ; Nippon Kayaku); mouse monoclonal anti-TLR4 (HTA125) and anti-TLR2 (TL2.1) antibodies (Hycult Biotechnology); mouse monoclonal anti-CD14 (MY4) antibody (Beckman Coulter); rabbit polyclonal anti-p44/42, anti-p38, anti-phosphorylated p38, anti-phosphorylated IκB-α, mouse monoclonal anti-phosphorylated p44/42 antibodies (Cell Signaling); mouse monoclonal anti-JUN-N-terminal protein kinase (JNK) (D-2), anti-phosphorylated JNK (G-7), anti-IκB-α (H-4) antibodies (Santa Cruz Biotechnology); and mouse monoclonal anti-β-actin antibody (Abcam).

Techniques: Activation Assay, Luciferase, Transfection, Activity Assay

Journal: Journal of Inflammation (London, England)

Article Title: Salivary histatin 3 inhibits heat shock cognate protein 70-mediated inflammatory cytokine production through toll-like receptors in human gingival fibroblasts

doi: 10.1186/1476-9255-11-4

Figure Lengend Snippet: The effect of DSG on NF-κB-dependent activation by HSC70 through TLR2 and TLR4 in the presence of histatin 3. One day after transfection with pIgκB-Luc in 293-TLR4/MD2-CD14 (A) and 293-TLR2/CD14 (B) cells, the cells were stimulated with HSC70 with or without histatin 3 in the presence or absence of DSG for 6 h. Cell lysates prepared from the stimulated cells were analyzed by luciferase assays. The values are shown as fold induction of the standardized luciferase activity over the unstimulated control (none, DSG (-)). Bars represent the means and range of duplicate samples. *, P < 0.05; **, P < 0.01, ***, P < 0.001 versus stimulation with HSC70 alone. †††, P < 0.001.

Article Snippet: ; Nippon Kayaku); mouse monoclonal anti-TLR4 (HTA125) and anti-TLR2 (TL2.1) antibodies (Hycult Biotechnology); mouse monoclonal anti-CD14 (MY4) antibody (Beckman Coulter); rabbit polyclonal anti-p44/42, anti-p38, anti-phosphorylated p38, anti-phosphorylated IκB-α, mouse monoclonal anti-phosphorylated p44/42 antibodies (Cell Signaling); mouse monoclonal anti-JUN-N-terminal protein kinase (JNK) (D-2), anti-phosphorylated JNK (G-7), anti-IκB-α (H-4) antibodies (Santa Cruz Biotechnology); and mouse monoclonal anti-β-actin antibody (Abcam).

Techniques: Activation Assay, Transfection, Luciferase, Activity Assay

Journal: Journal of Inflammation (London, England)

Article Title: Salivary histatin 3 inhibits heat shock cognate protein 70-mediated inflammatory cytokine production through toll-like receptors in human gingival fibroblasts

doi: 10.1186/1476-9255-11-4

Figure Lengend Snippet: The effect of anti-TLR antibodies on HSC70-stimulated inflammatory cytokine production in HGFs. (A) IL-6 and (B) IL-8 production with LPS or HSC70 in the presence of anti-TLR2 and anti-TLR4 antibodies in HGFs. HGFs were cultured for 1 h after the addition of the antibodies and were then stimulated with LPSs or HSC70 for 24 h, and ELISAs for IL-6 and IL-8 were performed on the cultured media. (C) LPS- and HSC70-stimulated IL-6 and (D) IL-8 production in the presence of anti-CD14 antibody in HGFs. Assays were performed as described in the “Methods.” IgG served as the antibody control. Bars represent the means and range of duplicate samples. ***, P < 0.001 versus stimulation with LPS or HSC70 alone.

Article Snippet: ; Nippon Kayaku); mouse monoclonal anti-TLR4 (HTA125) and anti-TLR2 (TL2.1) antibodies (Hycult Biotechnology); mouse monoclonal anti-CD14 (MY4) antibody (Beckman Coulter); rabbit polyclonal anti-p44/42, anti-p38, anti-phosphorylated p38, anti-phosphorylated IκB-α, mouse monoclonal anti-phosphorylated p44/42 antibodies (Cell Signaling); mouse monoclonal anti-JUN-N-terminal protein kinase (JNK) (D-2), anti-phosphorylated JNK (G-7), anti-IκB-α (H-4) antibodies (Santa Cruz Biotechnology); and mouse monoclonal anti-β-actin antibody (Abcam).

Techniques: Cell Culture

Journal: PLoS ONE

Article Title: miR-1915 and miR-1225-5p Regulate the Expression of CD133, PAX2 and TLR2 in Adult Renal Progenitor Cells

doi: 10.1371/journal.pone.0068296

Figure Lengend Snippet: ( A ) TLR2 expression levels were analyzed by real-time PCR following transfection with 25 nM miR-1225-5p mimic. Increasing the amount of miR-1225-5p within ARPCs resulted in a 1.3 fold reduction of TLR2 mRNA levels 24 hours after transfection. Expression data were normalized on the housekeeping gene β-actin. Data are representative of three independent experiments (means ± SEM), *p<0.01. ( B–C ) TLR2 protein expression after transfection with 50 nM miR-1225-5p mimic. A strong reduction of TLR2 in ARPC was found after 3 days from transfection with miR-1225-5p mimic, as shown by immunofluorescence staining. To-pro-3 counterstains nuclei (blue). Original view X63.

Article Snippet: The cells were blocked for 1 h (BSA in PBS, pH 7.4) and then incubated with a mouse anti-human TLR2 mAb (Hycult Biotechnology) and a mouse anti-human CD133 mAb (Miltenyi Biotec) overnight at 4°C.

Techniques: Expressing, Real-time Polymerase Chain Reaction, Transfection, Immunofluorescence, Staining

Journal: BMC Dermatology

Article Title: Toll-like receptor 2 activation and comedogenesis: implications for the pathogenesis of acne

doi: 10.1186/1471-5945-13-10

Figure Lengend Snippet: TLR Expression in vitro and in vivo . A) TLR2 expression was assessed by RT-PCR in cDNA prepared from HaCaT keratinocytes (first lane), NHEK (second lane) or freshly-isolated human sebaceous glands (third lane, representative of ten different individuals). A no-template cDNA control was included (fourth lane). B) TLR2 antisera were used to evaluate receptor localisation in skin isolated from ten individuals and representative expression patterns are shown, with and without a blocking peptide (pep). Images were captured at 10 × original magnification.

Article Snippet: Antibodies raised against human TLR2 and TLR4 were obtained from HyCult Biotechnology bv (Uden, Netherlands) and used at 100 μg/ml to block the effect of the PAMPs.

Techniques: Expressing, In Vitro, In Vivo, Reverse Transcription Polymerase Chain Reaction, Isolation, Blocking Assay

Journal: BMC Dermatology

Article Title: Toll-like receptor 2 activation and comedogenesis: implications for the pathogenesis of acne

doi: 10.1186/1471-5945-13-10

Figure Lengend Snippet: TLR activation results in IL-1α release and NF-κB activation. A) Preconfluent NHEK were treated with PGN in the presence or absence of a TLR2 neutralising antibody, and levels of IL-1α release determined by ELISA. B) Three separate cultures of preconfluent NHEK were harvested following 90 min exposure to PGN, and IκB expression relative to β-actin was determined by Western blotting.

Article Snippet: Antibodies raised against human TLR2 and TLR4 were obtained from HyCult Biotechnology bv (Uden, Netherlands) and used at 100 μg/ml to block the effect of the PAMPs.

Techniques: Activation Assay, Enzyme-linked Immunosorbent Assay, Expressing, Western Blot

Journal: BMC Dermatology

Article Title: Toll-like receptor 2 activation and comedogenesis: implications for the pathogenesis of acne

doi: 10.1186/1471-5945-13-10

Figure Lengend Snippet: TLR activation promotes hypercornification in isolated human sebaceous glands maintained ex vivo . A) Histological sections of freshly-isolated and IL-1α treated sebaceous glands are shown, along with an example of a comedone in situ . B) Morphology representative of 3 experiments showing sebaceous glands treated with vehicle only (control), TLR2 (LTA and PGN) or TLR4 agonists (LPS), or TLR agonists pre-incubated with a blocking antibody (block). All images are shown at 10 × original magnification.

Article Snippet: Antibodies raised against human TLR2 and TLR4 were obtained from HyCult Biotechnology bv (Uden, Netherlands) and used at 100 μg/ml to block the effect of the PAMPs.

Techniques: Activation Assay, Isolation, Ex Vivo, In Situ, Incubation, Blocking Assay

Journal: Medical Oncology (Northwood, London, England)

Article Title: Evidence of a role for interleukin-6 in anoikis resistance in oral squamous cell carcinoma

doi: 10.1007/s12032-022-01664-5

Figure Lengend Snippet: Cytokine production in PGK, DOK and SCC-4 cells. PGK, DOK and SCC-4 were untreated or stimulated for 24 h with TLR2 agonists: Pam3CSK4 (0.2 μg/ml), Pam2CSK4 (0.2 μg/ml) or TLR4 agonist LPS (0.5 μg/ml). Levels of ( A ) IL-6, ( B ) IL-11, ( C ) IL-8 and ( D ) TNF-α in the culture supernatants were determined by ELISA. Results were plotted using Graphpad prism 8. Data shown are representative of at least three independent experiments. Values represent the mean ± S.E.M of three independent experiments. Statistical analysis was performed using one-way ANOVA with a post hoc Tukey’s test to compare mean values between untreated and treated groups within the cell line, * p < 0.05; ** p < 0.01; *** p < 0.001

Article Snippet: Antibody information: Human anti-IL-6Ra (MAB227), Mouse monoclonal IgG 1 , clone 17506 (R&D systems); Human IL-6 (MAB2061), 1936 (R&D systems); Isotype Control (MAB004), mouse monoclonal IgG 2B , clone 20116 (R&D systems); Human TLR2 antibody (HM1054), mouse monoclonal IgG 1, Clone T2.5 (Hycult Biotech).

Techniques: Enzyme-linked Immunosorbent Assay

Journal: Medical Oncology (Northwood, London, England)

Article Title: Evidence of a role for interleukin-6 in anoikis resistance in oral squamous cell carcinoma

doi: 10.1007/s12032-022-01664-5

Figure Lengend Snippet: The effect of TLR2 neutralising antibody on IL-6 production in SCC-4 cells. ( A ) DOK and ( B ) SCC-4 cells were untreated or pre-treated with anti-TLR2 Ab (10 μg/ml) for 1 h, prior to stimulation with Pam2CSK4 (0.2 μg/ml) and Pam3CSK4 (0.2 μg/ml) for 24 h. Culture supernatants were analysed for IL-6 secretion by ELISA. Results were plotted using Graphpad prism 8. Values represent the mean ± S.E.M of three independent experiments. Statistical analysis was performed using one-way ANOVA with a post hoc Dunnett’s test to compare mean values among the groups, * p < 0.05; ** p < 0.01; *** p < 0.001

Article Snippet: Antibody information: Human anti-IL-6Ra (MAB227), Mouse monoclonal IgG 1 , clone 17506 (R&D systems); Human IL-6 (MAB2061), 1936 (R&D systems); Isotype Control (MAB004), mouse monoclonal IgG 2B , clone 20116 (R&D systems); Human TLR2 antibody (HM1054), mouse monoclonal IgG 1, Clone T2.5 (Hycult Biotech).

Techniques: Enzyme-linked Immunosorbent Assay

Journal: Medical Oncology (Northwood, London, England)

Article Title: Evidence of a role for interleukin-6 in anoikis resistance in oral squamous cell carcinoma

doi: 10.1007/s12032-022-01664-5

Figure Lengend Snippet: TLR2 and TLR6 expression levels in SCC-4 cells and DOK cells. DOK and SCC-4 cells were left untreated or pre-treated with anti-TLR2 Ab (10 μg/ml) for 1 h, prior to stimulation with Pam3CSK4 (0.05 μg/ml) or Pam2CSK4 (0.05 μg/ml) for 24 h. Lysates were collected and samples were run on 12% SDS-PAGE gel. Western Blot were probed with anti-TLR2 and anti-TLR6 antibodies with anti-GAPDH used as a loading control. Experiment was done in triplicate. Densitometric analysis was performed using image lab software on TLR2, TLR6 and GAPDH blots. Sample blots for ( A ) DOK and ( B ) SCC-4 and collated densitometry data for ( C ) TLR2 and ( D ) TLR6 are presented. Results were plotted using Graphpad prism 8. Values represent the mean ± S.E.M of three independent experiments. Statistical analysis was performed using two-way ANOVA with a post hoc Sidak’s test to compare mean values among the groups, * p < 0.05; ** p < 0.01; *** p < 0.001

Article Snippet: Antibody information: Human anti-IL-6Ra (MAB227), Mouse monoclonal IgG 1 , clone 17506 (R&D systems); Human IL-6 (MAB2061), 1936 (R&D systems); Isotype Control (MAB004), mouse monoclonal IgG 2B , clone 20116 (R&D systems); Human TLR2 antibody (HM1054), mouse monoclonal IgG 1, Clone T2.5 (Hycult Biotech).

Techniques: Expressing, SDS Page, Western Blot, Software

Journal: Medical Oncology (Northwood, London, England)

Article Title: Evidence of a role for interleukin-6 in anoikis resistance in oral squamous cell carcinoma

doi: 10.1007/s12032-022-01664-5

Figure Lengend Snippet: Survival rate in anoikis assays DOK and SCC-4 cells treated with rhIL-6, IL-6 neutralising antibody, IL-6Ra antibody, IL-6Ra + rhIL-6, Pam2CSK4, anti-TLR2 neutralising antibody + Pam2CSK4, and anti-TLR2 neutralising antibody + Pam2CSK4 for 24 h. Tissue culture plates were coated with 200 μl poly-(hydroxyethyl methacrylic) acid (poly-HEMA) thus inhibiting the ability of the cells to attach to the tissue culture plastic. ( A ) DOK and ( B ) SCC-4 cells were then seeded at 100,000 cells/well and allowed to attach. For those cells to be treated with 30 ng/ml rhIL-6, 10 μg/ml IL-6 neutralising antibody, 40 μg/ml IL-6Ra antibody, 40 μg/ml IL-6Ra + 30 ng/ml rhIL-6, 0.2 μg/ml Pam2CSK4, and 10 μg/ml anti-TLR2 neutralising antibody + 0.2 μg/ml Pam2CSK4, complete medium was used as control, as well as medium containing 10 μg/ml isotype IgG antibody. After 24 h, 50 μl of alamarBlue ® dye was added to each well and incubated again at 37 °C/5% CO 2 for 3.5 h. SCC-4 cells demonstrated significant resistance to anoikis (% survival) when treated with 30 ng/ml rhIL-6 and 0.2 μg/ml Pam2CSK4 compared to the control group. There is a100% survival of adherent cells in the uncoated plates. Fluorescence was measured using a SpectraMax Gemini plate reader at excitation wavelength 544 nm and emission wavelength 590 nm. Results were plotted using Graphpad prism 8. Results represent at least three experiments performed in duplicate (mean ± SEM ( n )). Statistical analysis was performed one-way ANOVA with a post hoc Tukey’s test to compare mean values among the groups, * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001

Article Snippet: Antibody information: Human anti-IL-6Ra (MAB227), Mouse monoclonal IgG 1 , clone 17506 (R&D systems); Human IL-6 (MAB2061), 1936 (R&D systems); Isotype Control (MAB004), mouse monoclonal IgG 2B , clone 20116 (R&D systems); Human TLR2 antibody (HM1054), mouse monoclonal IgG 1, Clone T2.5 (Hycult Biotech).

Techniques: Incubation, Fluorescence

Journal: Shock (Augusta, Ga.)

Article Title: TLR2 on bone marrow and non-bone marrow derived cells regulates inflammation and organ injury in cooperation with TLR4 during resuscitated hemorrhagic shock

doi: 10.1097/SHK.0000000000000650

Figure Lengend Snippet: C57BL/6 mice or TLR2 KO mice underwent H/R procedure and were sacrificed at 6h or 20h TP. (A) IL-6, (B) IP10 (C) KC, (D) MIG, (E) MCP-1, (F) ALT or (G) AST plasma parameters were displayed. n=6–8 per experimental group, line and * represent p<0.05, Data represent mean+/−SEM.

Article Snippet: Antibodies Anti-TLR2 mouse IgG1 mAb T2.5 was from HyCult Biotechnology (Uden, Neitherlands) and has been described previously ( 23 ).

Techniques:

Journal: Shock (Augusta, Ga.)

Article Title: TLR2 on bone marrow and non-bone marrow derived cells regulates inflammation and organ injury in cooperation with TLR4 during resuscitated hemorrhagic shock

doi: 10.1097/SHK.0000000000000650

Figure Lengend Snippet: using TLR2 KO and C57/Bl6 mice WT/WT, WT/KO, KO/KO and KO/WT recipient/donor chimeric mice combinations were generated. Chimeric mice underwent H/R procedure and were sacrificed at 20h TP. (A) ALT, (B) IL-6, (C) IP10, (D) MCP-1, and (E) MIG plasma parameters were displayed. n=6–8 per experimental group, * p<0.05, Data represent mean+/−SEM.

Article Snippet: Antibodies Anti-TLR2 mouse IgG1 mAb T2.5 was from HyCult Biotechnology (Uden, Neitherlands) and has been described previously ( 23 ).

Techniques: Generated

Journal: Shock (Augusta, Ga.)

Article Title: TLR2 on bone marrow and non-bone marrow derived cells regulates inflammation and organ injury in cooperation with TLR4 during resuscitated hemorrhagic shock

doi: 10.1097/SHK.0000000000000650

Figure Lengend Snippet: C57/BL6 mice were injected i.p. with thioglycollate and peritoneal macrophages were harvested. Macrophages were stimulated in vitro with (A) PAM3CSK (1μg/ml), (B) ultrapure LPS (2ng/ml) or (C) PBS for 15min, 30min or 45min after incubation with different mAB (T2.5, 5E3, T2.5+5E3 or control, 10μg/ml). Translocation of NFκB p65 were measured and displayed in arbitrary units (n=8 or 24) per experimental group. Blue line represent mean of un-stimulated control (49+/−4 AU). Black lines represent *p<0.05 for 30min TP comparing groups, data represent mean+/−SD.

Article Snippet: Antibodies Anti-TLR2 mouse IgG1 mAb T2.5 was from HyCult Biotechnology (Uden, Neitherlands) and has been described previously ( 23 ).

Techniques: Injection, In Vitro, Incubation, Translocation Assay

Journal: Shock (Augusta, Ga.)

Article Title: TLR2 on bone marrow and non-bone marrow derived cells regulates inflammation and organ injury in cooperation with TLR4 during resuscitated hemorrhagic shock

doi: 10.1097/SHK.0000000000000650

Figure Lengend Snippet: (A) H&E staining of liver tissue sections from monoclonal TLR2 Antibody T2.5 (labeled as “TLR2”) or corresponding isotype control mAB (labeled as “control”) pretreated animals are displayed. Mice underwent H/R or sham procedure (A1 – A4). (B) H&E staining of paraformaldehyde-fixed and paraffin-embedded lung from anti-TLR2 or isotype control mAB are displayed. Mice underwent H/R or sham procedure (B1 – B4). Plasma ALT levels of untreated control animals (white bars) or ALT plasma levels of isotype control mAB (black bars) or anti-TLR2 mAB (grey bars) treated mice after H/R procedure are displayed. Blue line represent mean of sham group plasma levels of mice receiving isotype control mAB (C). Average of lung injury using interstitial edema, alveolar edema, hemorrhage, and neutrophil infiltrates as marker (scale in arbitrary units from 0=min to 16=max) White bar displays sham mice receiving isotype control or anti TLR2 mAB, grey and black bars displays corresponding H/R group (D). N=6–16 per experimental group, *p<0.05 (C) or *p<0.01 (D), Data represent mean+/−SEM, scale bar 100μm

Article Snippet: Antibodies Anti-TLR2 mouse IgG1 mAb T2.5 was from HyCult Biotechnology (Uden, Neitherlands) and has been described previously ( 23 ).

Techniques: Staining, Labeling, Marker

Journal: Shock (Augusta, Ga.)

Article Title: TLR2 on bone marrow and non-bone marrow derived cells regulates inflammation and organ injury in cooperation with TLR4 during resuscitated hemorrhagic shock

doi: 10.1097/SHK.0000000000000650

Figure Lengend Snippet: C57/Bl6 mice were pretreated with 100μg of control mAB or T2.5 mAB and/or 5E3 mAB 30min before H/R procedure. (A) ALT plasma levels in U/l are displayed, (B–F) IL-6, MIG, MCP-1, IP-10 and KC are shown at two different time points (6h and 20h TP). significant differences between two groups are marked by a line on top of the bars. Blue lines illustrate mean plasma levels of sham control mice, dotted line illustrate uninjured control plasma levels. (n=6–14 per experimental group, *p<0.05, Data represent mean+/−SEM)

Article Snippet: Antibodies Anti-TLR2 mouse IgG1 mAb T2.5 was from HyCult Biotechnology (Uden, Neitherlands) and has been described previously ( 23 ).

Techniques:

Journal: Shock (Augusta, Ga.)

Article Title: TLR2 on bone marrow and non-bone marrow derived cells regulates inflammation and organ injury in cooperation with TLR4 during resuscitated hemorrhagic shock

doi: 10.1097/SHK.0000000000000650

Figure Lengend Snippet: C57BL/6 mice or TLR2 KO mice underwent H/R procedure and were sacrificed at 6h or 20h TP. (A) IL-6, (B) IP10 (C) KC, (D) MIG, (E) MCP-1, (F) ALT or (G) AST plasma parameters were displayed. n=6–8 per experimental group, line and * represent p<0.05, Data represent mean+/−SEM.

Article Snippet: Anti-TLR2 mouse IgG1 mAb T2.5 was from HyCult Biotechnology (Uden, Neitherlands) and has been described previously ( 23 ).

Techniques:

Journal: Shock (Augusta, Ga.)

Article Title: TLR2 on bone marrow and non-bone marrow derived cells regulates inflammation and organ injury in cooperation with TLR4 during resuscitated hemorrhagic shock

doi: 10.1097/SHK.0000000000000650

Figure Lengend Snippet: using TLR2 KO and C57/Bl6 mice WT/WT, WT/KO, KO/KO and KO/WT recipient/donor chimeric mice combinations were generated. Chimeric mice underwent H/R procedure and were sacrificed at 20h TP. (A) ALT, (B) IL-6, (C) IP10, (D) MCP-1, and (E) MIG plasma parameters were displayed. n=6–8 per experimental group, * p<0.05, Data represent mean+/−SEM.

Article Snippet: Anti-TLR2 mouse IgG1 mAb T2.5 was from HyCult Biotechnology (Uden, Neitherlands) and has been described previously ( 23 ).

Techniques: Generated

Journal: Shock (Augusta, Ga.)

Article Title: TLR2 on bone marrow and non-bone marrow derived cells regulates inflammation and organ injury in cooperation with TLR4 during resuscitated hemorrhagic shock

doi: 10.1097/SHK.0000000000000650

Figure Lengend Snippet: C57/BL6 mice were injected i.p. with thioglycollate and peritoneal macrophages were harvested. Macrophages were stimulated in vitro with (A) PAM3CSK (1μg/ml), (B) ultrapure LPS (2ng/ml) or (C) PBS for 15min, 30min or 45min after incubation with different mAB (T2.5, 5E3, T2.5+5E3 or control, 10μg/ml). Translocation of NFκB p65 were measured and displayed in arbitrary units (n=8 or 24) per experimental group. Blue line represent mean of un-stimulated control (49+/−4 AU). Black lines represent *p<0.05 for 30min TP comparing groups, data represent mean+/−SD.

Article Snippet: Anti-TLR2 mouse IgG1 mAb T2.5 was from HyCult Biotechnology (Uden, Neitherlands) and has been described previously ( 23 ).

Techniques: Injection, In Vitro, Incubation, Translocation Assay

Journal: Shock (Augusta, Ga.)

Article Title: TLR2 on bone marrow and non-bone marrow derived cells regulates inflammation and organ injury in cooperation with TLR4 during resuscitated hemorrhagic shock

doi: 10.1097/SHK.0000000000000650

Figure Lengend Snippet: (A) H&E staining of liver tissue sections from monoclonal TLR2 Antibody T2.5 (labeled as “TLR2”) or corresponding isotype control mAB (labeled as “control”) pretreated animals are displayed. Mice underwent H/R or sham procedure (A1 – A4). (B) H&E staining of paraformaldehyde-fixed and paraffin-embedded lung from anti-TLR2 or isotype control mAB are displayed. Mice underwent H/R or sham procedure (B1 – B4). Plasma ALT levels of untreated control animals (white bars) or ALT plasma levels of isotype control mAB (black bars) or anti-TLR2 mAB (grey bars) treated mice after H/R procedure are displayed. Blue line represent mean of sham group plasma levels of mice receiving isotype control mAB (C). Average of lung injury using interstitial edema, alveolar edema, hemorrhage, and neutrophil infiltrates as marker (scale in arbitrary units from 0=min to 16=max) White bar displays sham mice receiving isotype control or anti TLR2 mAB, grey and black bars displays corresponding H/R group (D). N=6–16 per experimental group, *p<0.05 (C) or *p<0.01 (D), Data represent mean+/−SEM, scale bar 100μm

Article Snippet: Anti-TLR2 mouse IgG1 mAb T2.5 was from HyCult Biotechnology (Uden, Neitherlands) and has been described previously ( 23 ).

Techniques: Staining, Labeling, Marker

Journal: Shock (Augusta, Ga.)

Article Title: TLR2 on bone marrow and non-bone marrow derived cells regulates inflammation and organ injury in cooperation with TLR4 during resuscitated hemorrhagic shock

doi: 10.1097/SHK.0000000000000650

Figure Lengend Snippet: C57/Bl6 mice were pretreated with 100μg of control mAB or T2.5 mAB and/or 5E3 mAB 30min before H/R procedure. (A) ALT plasma levels in U/l are displayed, (B–F) IL-6, MIG, MCP-1, IP-10 and KC are shown at two different time points (6h and 20h TP). significant differences between two groups are marked by a line on top of the bars. Blue lines illustrate mean plasma levels of sham control mice, dotted line illustrate uninjured control plasma levels. (n=6–14 per experimental group, *p<0.05, Data represent mean+/−SEM)

Article Snippet: Anti-TLR2 mouse IgG1 mAb T2.5 was from HyCult Biotechnology (Uden, Neitherlands) and has been described previously ( 23 ).

Techniques: