mouse anti human tlr4 pe  (Bio-Rad)

Journal: PLoS ONE

Article Title: Porphyromonas Gingivalis and E-coli Induce Different Cytokine Production Patterns in Pregnant Women

doi: 10.1371/journal.pone.0086355

Figure Lengend Snippet: Leukocytes were selected in the forward-sidescatter plot (fig. 1A) and copied to a sidescatter-CD14 plot. Monocytes (CD14 positive cells), granulocytes (CD14 negative cells with high SSC) and lymphocytes (CD14 negative cells with low SSC) were gated (fig. 1B). CD14 positive cells were copied to a TLR2/TLR4 plot. Using the isotype control sample, gates were set in the TLR2/TLR4 plot so that at least 99% of the isotype controls were negative for TLR2/TLR4 expression (fig. 1C). This gate was then used to identify the percentages of TLR4/TLR2 double positive, TLR2 single and TLR4 single positive monocytes as well as their mean fluorescence intensity (MFI), in the antibody incubated samples (fig. 1D).

Article Snippet: Immediately after sampling, 500 µl of whole blood was mixed with 500 µl of RPMI and incubated with PerCp–labeled mouse-anti-human-CD14 (clone TüK4; Invitrogen Corporation, Breda, The Netherlands) together with FITC-labeled mouse-anti-human-TLR2 (clone TL2.1; eBioscience, Breda, The Netherlands) and PE-labeled mouse-anti-human-TLR4 (clone HTA 125; eBiosciences), or with anti-CD14 together with TLR2 and TLR4 isotype controls for 30 minutes at room temperature (RT) in the dark.

Techniques: Expressing, Fluorescence, Incubation

Journal: PLoS ONE

Article Title: Porphyromonas Gingivalis and E-coli Induce Different Cytokine Production Patterns in Pregnant Women

doi: 10.1371/journal.pone.0086355

Figure Lengend Snippet: Expression of TLR2 and TLR4 on monocytes of pregnant (open squares) and non-pregnant women (black squares). A: Percentage of TLR2 positive monocytes (left graph) and mean fluorescent intensity of TLR2 staining of monocytes (right graph). B: Percentage of TLR4 positive monocytes (left graph) and mean fluorescent intensity of TLR4 staining of monocytes (right graph). *: significantly increased vs pregnant women (student’s T test, p<0.05).

Article Snippet: Immediately after sampling, 500 µl of whole blood was mixed with 500 µl of RPMI and incubated with PerCp–labeled mouse-anti-human-CD14 (clone TüK4; Invitrogen Corporation, Breda, The Netherlands) together with FITC-labeled mouse-anti-human-TLR2 (clone TL2.1; eBioscience, Breda, The Netherlands) and PE-labeled mouse-anti-human-TLR4 (clone HTA 125; eBiosciences), or with anti-CD14 together with TLR2 and TLR4 isotype controls for 30 minutes at room temperature (RT) in the dark.

Techniques: Expressing, Staining

Journal: Journal of Immunology Research

Article Title: 43 kDa Glycoprotein (gp43) from Paracoccidioides brasiliensis Induced IL-17A and PGE2 Production by Human Polymorphonuclear Neutrophils: Involvement of TLR2 and TLR4

doi: 10.1155/2019/1790908

Figure Lengend Snippet: Percentage of TLR2 (a) and TLR4 (b) expression on the surface of CD16+ human polymorphonuclear neutrophils (PMNs). PMNs were incubated in the absence (control) or presence of gp43 (20 ng/ml) for 4 h, and PMNs were analyzed using flow cytometry. Results are presented as percentage of PMNs expressing TLR2 and TLR4 using box-and-whiskers (min to max) graphs with data from 7 women tested, considering p < 0.05. Statistically significant differences between groups are indicated as follows: ∗ p < 0.05 and ∗∗ p < 0.01. (c) Representative experiment showing the expression of PMN (control culture) CD16 PerCP+, TLR2 FITC+, and TLR4 PE+, and histograms display the percentage of TLR2 and TLR4 blockade (red line) and the remaining positive receptor expression (gray lines).

Article Snippet: PMNs were then incubated with PerCP anti-human CD16 antibody, FITC anti-human CD282 (TLR2) antibody (both antibodies from BioLegend, San Diego, CA, USA), and PE mouse anti-human TLR4 antibody (BD Pharmingen™, Franklin Lakes, NJ, USA) for 20 minutes at 4°C in the dark.

Techniques: Expressing, Incubation, Flow Cytometry

Journal: Journal of Immunology Research

Article Title: 43 kDa Glycoprotein (gp43) from Paracoccidioides brasiliensis Induced IL-17A and PGE2 Production by Human Polymorphonuclear Neutrophils: Involvement of TLR2 and TLR4

doi: 10.1155/2019/1790908

Figure Lengend Snippet: Involvement of TLR2 and TLR4 in PGE2 (a) and LTB4 (b) production by human polymorphonuclear neutrophils (PMNs) stimulated with gp43 from P. brasiliensis . Cells were incubated in the absence or presence of anti-TLR2 and anti-TLR4 antibodies, individually or in combination for 2 h. PMNs were then stimulated or not with gp43 (20 ng/ml) during 4 h, and eicosanoid concentrations (pg/ml) were measured by competitive ELISA in the culture supernatants. Results are expressed using a box-and-whiskers (min to max) graph with data from 7 women tested, considering p < 0.05 (a) or mean ± SD (b), considering p < 0.05. Statistically significant differences between groups are indicated as follows: ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001.

Article Snippet: PMNs were then incubated with PerCP anti-human CD16 antibody, FITC anti-human CD282 (TLR2) antibody (both antibodies from BioLegend, San Diego, CA, USA), and PE mouse anti-human TLR4 antibody (BD Pharmingen™, Franklin Lakes, NJ, USA) for 20 minutes at 4°C in the dark.

Techniques: Incubation, Competitive ELISA

Journal: Journal of Immunology Research

Article Title: 43 kDa Glycoprotein (gp43) from Paracoccidioides brasiliensis Induced IL-17A and PGE2 Production by Human Polymorphonuclear Neutrophils: Involvement of TLR2 and TLR4

doi: 10.1155/2019/1790908

Figure Lengend Snippet: Percentage of human polymorphonuclear neutrophils (PMNs) producing intracytoplasmic IL-17A (a) and IL-4 (b) after stimulation or not with gp43 from P. brasiliensis and the involvement of TLR2 and TLR4. Cells were incubated in the absence or presence of anti-TLR2 and anti-TLR4 antibodies, individually or in combination, for 2 h. PMNs were then stimulated or not with gp43 (20 ng/ml) during 4 h and intracellular IL-4 and IL-17 were analyzed by flow cytometry. Results are presented as the percentage of PMNs expressing IL-17A (a) and IL-4 (b) using box-and-whiskers (min to max) graphs with data from 7 women tested, considering p < 0.05. (a) Statistically significant differences between groups are indicated as follows: ∗ p < 0.05 and ∗∗ p < 0.01.

Article Snippet: PMNs were then incubated with PerCP anti-human CD16 antibody, FITC anti-human CD282 (TLR2) antibody (both antibodies from BioLegend, San Diego, CA, USA), and PE mouse anti-human TLR4 antibody (BD Pharmingen™, Franklin Lakes, NJ, USA) for 20 minutes at 4°C in the dark.

Techniques: Incubation, Flow Cytometry, Expressing

Journal: Journal of Immunology Research

Article Title: 43 kDa Glycoprotein (gp43) from Paracoccidioides brasiliensis Induced IL-17A and PGE2 Production by Human Polymorphonuclear Neutrophils: Involvement of TLR2 and TLR4

doi: 10.1155/2019/1790908

Figure Lengend Snippet: Involvement of TLR2 and TLR4 in the production of IL-6 (a), IL-10 (b), IL-12 (c), IFN- γ (d), and TNF- α (e) by human polymorphonuclear neutrophils (PMN) stimulated or not with gp43 from P. brasiliensis . Cells were incubated in the absence or presence of anti-TLR2 and anti-TLR4 antibodies, individually or in combination, for 2 h. PMNs were then stimulated or not with gp43 (20 ng/ml) during 4 h, and cytokine concentrations (pg/ml) were measured in the supernatant culture. Results are expressed as median using data from 7 women tested, considering p < 0.05.

Article Snippet: PMNs were then incubated with PerCP anti-human CD16 antibody, FITC anti-human CD282 (TLR2) antibody (both antibodies from BioLegend, San Diego, CA, USA), and PE mouse anti-human TLR4 antibody (BD Pharmingen™, Franklin Lakes, NJ, USA) for 20 minutes at 4°C in the dark.

Techniques: Incubation

Journal: Biochemistry and Biophysics Reports

Article Title: The effect of probiotic bacteria on toll-like receptor-2 and -4 expression by spermatozoa in couples with unexplained recurrent spontaneous abortion

doi: 10.1016/j.bbrep.2022.101390

Figure Lengend Snippet: Flow cytometric analysis of TLR2 and TLR4 expression in Fluorescence Minus One (FMO) control, untreated and probiotic treated spermatozoa groups.

Article Snippet: Untreated and probiotic treated spermatozoa were stained with phycoerythrin (PE) mouse anti-human TLR4 (BD pharmingen, USA) and Fluorescein-5-isothiocyanate (FITC) mouse anti-human TLR2 (Southern Biotech).

Techniques: Expressing, Fluorescence

Journal: Biochemistry and Biophysics Reports

Article Title: The effect of probiotic bacteria on toll-like receptor-2 and -4 expression by spermatozoa in couples with unexplained recurrent spontaneous abortion

doi: 10.1016/j.bbrep.2022.101390

Figure Lengend Snippet: Total mean of the TLR2 and TLR4 variables in the type groups.

Article Snippet: Untreated and probiotic treated spermatozoa were stained with phycoerythrin (PE) mouse anti-human TLR4 (BD pharmingen, USA) and Fluorescein-5-isothiocyanate (FITC) mouse anti-human TLR2 (Southern Biotech).

Techniques:

Journal: Biochemistry and Biophysics Reports

Article Title: The effect of probiotic bacteria on toll-like receptor-2 and -4 expression by spermatozoa in couples with unexplained recurrent spontaneous abortion

doi: 10.1016/j.bbrep.2022.101390

Figure Lengend Snippet: Total mean of the TLR2 and TLR4 variables in the treatment groups.

Article Snippet: Untreated and probiotic treated spermatozoa were stained with phycoerythrin (PE) mouse anti-human TLR4 (BD pharmingen, USA) and Fluorescein-5-isothiocyanate (FITC) mouse anti-human TLR2 (Southern Biotech).

Techniques:

Journal: Biochemistry and Biophysics Reports

Article Title: The effect of probiotic bacteria on toll-like receptor-2 and -4 expression by spermatozoa in couples with unexplained recurrent spontaneous abortion

doi: 10.1016/j.bbrep.2022.101390

Figure Lengend Snippet: Profile plot of type (fertile, RSA) versus treatment (untreated [UT], probiotic treated [PRO]). Results show significant interaction between type (fertile, RSA) and probiotic treatment for TLR2 and TLR4.

Article Snippet: Untreated and probiotic treated spermatozoa were stained with phycoerythrin (PE) mouse anti-human TLR4 (BD pharmingen, USA) and Fluorescein-5-isothiocyanate (FITC) mouse anti-human TLR2 (Southern Biotech).

Techniques:

Journal: Biochemistry and Biophysics Reports

Article Title: The effect of probiotic bacteria on toll-like receptor-2 and -4 expression by spermatozoa in couples with unexplained recurrent spontaneous abortion

doi: 10.1016/j.bbrep.2022.101390

Figure Lengend Snippet: Results obtained from treatment comparison (untreated, probiotic treated).

Article Snippet: Untreated and probiotic treated spermatozoa were stained with phycoerythrin (PE) mouse anti-human TLR4 (BD pharmingen, USA) and Fluorescein-5-isothiocyanate (FITC) mouse anti-human TLR2 (Southern Biotech).

Techniques:

Journal: Biochemistry and Biophysics Reports

Article Title: The effect of probiotic bacteria on toll-like receptor-2 and -4 expression by spermatozoa in couples with unexplained recurrent spontaneous abortion

doi: 10.1016/j.bbrep.2022.101390

Figure Lengend Snippet: Results obtained from the type comparison (fertile, RSA).

Article Snippet: Untreated and probiotic treated spermatozoa were stained with phycoerythrin (PE) mouse anti-human TLR4 (BD pharmingen, USA) and Fluorescein-5-isothiocyanate (FITC) mouse anti-human TLR2 (Southern Biotech).

Techniques:

Journal: Immunity & Ageing : I & A

Article Title: Inflammation and cell-to-cell communication, two related aspects in frailty

doi: 10.1186/s12979-022-00306-8

Figure Lengend Snippet: The inflammatory markers highlight an increase in TLR2 + /calcein + lEVs and TLR4 + /calcein + lEVs in Fr subjects. A TLR2 + /calcein + lEVs show significantly higher plasma concentration in all Fr ( n = 20) compared to nFr ( n = 20) subjects (* p < 0.05). B A significant increase (* p < 0.05) was reported in male Fr ( n = 7) compared to Fit ( n = 11) subjects. C No differences were reported in the female Fr ( n = 13) versus nFr ( n = 9) group. D TLR4 + /calcein + lEVs show significantly increase higher plasma concentration in all Fr ( n = 20) compared to nFr ( n = 20) subjects (*** p < 0.001). E A significant increase (*** p < 0.001) was reported in male Fr ( n = 7) compared to nFr ( n = 11) subjects. F The female Fr ( n = 13) group shows a significant increase (** p < 0.01) in TLR4 + /calcein + lEVs versus nFr ( n = 9). Statistical analysis was performed using the Mann–Whitney U test (non-parametric) for comparisons between two groups using GraphPad Prism8 software. A p < 0.05 was considered statistically significant

Article Snippet: PE Mouse Anti-Human TLR4 (CD284) BD Biosciences , PerCP/Cyanine5.5 anti-human CD126 (IL-6Rα) BioLegend.

Techniques: Concentration Assay, MANN-WHITNEY, Software

Journal: Immunity & Ageing : I & A

Article Title: Inflammation and cell-to-cell communication, two related aspects in frailty

doi: 10.1186/s12979-022-00306-8

Figure Lengend Snippet: List of lEVs markers tested by flow cytometry analysis

Article Snippet: PE Mouse Anti-Human TLR4 (CD284) BD Biosciences , PerCP/Cyanine5.5 anti-human CD126 (IL-6Rα) BioLegend.

Techniques: Flow Cytometry

Journal: Frontiers in Immunology

Article Title: Toll-Interleukin 1 Receptor Domain-Containing Adaptor Protein 180L Single-Nucleotide Polymorphism Is Associated With Susceptibility to Recurrent Pneumococcal Lower Respiratory Tract Infections in Children

doi: 10.3389/fimmu.2018.01780

Figure Lengend Snippet: Primers and probes used in this study.

Article Snippet: PE anti-human TLR2 mAb (clone TL2.1, mouse IgG2a κ, RRID:AB_10667887), PE anti-human TLR4 mAb (clone HTA125, mouse IgG2a κ, RRID:AB_1603311), PE isotype control (clone eBM2a, mouse IgG2a κ, RRID:AB_470063), and APC anti-human CD14 mAb (clone 61D3, mouse IgG1, RRID:AB_10669167) were all purchased from eBioscience (THP Medical Products, Switzerland).

Techniques:

Journal: Frontiers in Immunology

Article Title: Toll-Interleukin 1 Receptor Domain-Containing Adaptor Protein 180L Single-Nucleotide Polymorphism Is Associated With Susceptibility to Recurrent Pneumococcal Lower Respiratory Tract Infections in Children

doi: 10.3389/fimmu.2018.01780

Figure Lengend Snippet: Relationship between TLR single-nucleotide polymorphisms and recurrent LRTI in children.

Article Snippet: PE anti-human TLR2 mAb (clone TL2.1, mouse IgG2a κ, RRID:AB_10667887), PE anti-human TLR4 mAb (clone HTA125, mouse IgG2a κ, RRID:AB_1603311), PE isotype control (clone eBM2a, mouse IgG2a κ, RRID:AB_470063), and APC anti-human CD14 mAb (clone 61D3, mouse IgG1, RRID:AB_10669167) were all purchased from eBioscience (THP Medical Products, Switzerland).

Techniques:

Journal: Frontiers in Immunology

Article Title: Toll-Interleukin 1 Receptor Domain-Containing Adaptor Protein 180L Single-Nucleotide Polymorphism Is Associated With Susceptibility to Recurrent Pneumococcal Lower Respiratory Tract Infections in Children

doi: 10.3389/fimmu.2018.01780

Figure Lengend Snippet: TLR-2 and TLR-4 expression on monocytes in peripheral blood.

Article Snippet: PE anti-human TLR2 mAb (clone TL2.1, mouse IgG2a κ, RRID:AB_10667887), PE anti-human TLR4 mAb (clone HTA125, mouse IgG2a κ, RRID:AB_1603311), PE isotype control (clone eBM2a, mouse IgG2a κ, RRID:AB_470063), and APC anti-human CD14 mAb (clone 61D3, mouse IgG1, RRID:AB_10669167) were all purchased from eBioscience (THP Medical Products, Switzerland).

Techniques: Expressing

Journal: Frontiers in Immunology

Article Title: Toll-Interleukin 1 Receptor Domain-Containing Adaptor Protein 180L Single-Nucleotide Polymorphism Is Associated With Susceptibility to Recurrent Pneumococcal Lower Respiratory Tract Infections in Children

doi: 10.3389/fimmu.2018.01780

Figure Lengend Snippet: Increased IL-6 release in LRTI children after whole-blood stimulation by Streptococcus pneumoniae and TLR2 and TLR4 ligands. The diagram shows scatter dot plots of IL-6 secretion in LRTI and healthy children after whole blood stimulation by five TLR ligands. Data are presented as mean and 95% CI (black bars). IL-6 secretion is expressed in picograms per milliliter for 10 4 monocytes. * P < 0.05; ** P < 0.01; and *** P < 0.001 by Mann–Whitney test. Abbreviations: TLR, toll-like receptor; LRTI, lower respiratory tract infections; n , number of children; IL-6, interleukin-6.

Article Snippet: PE anti-human TLR2 mAb (clone TL2.1, mouse IgG2a κ, RRID:AB_10667887), PE anti-human TLR4 mAb (clone HTA125, mouse IgG2a κ, RRID:AB_1603311), PE isotype control (clone eBM2a, mouse IgG2a κ, RRID:AB_470063), and APC anti-human CD14 mAb (clone 61D3, mouse IgG1, RRID:AB_10669167) were all purchased from eBioscience (THP Medical Products, Switzerland).

Techniques: MANN-WHITNEY

Journal: Frontiers in Immunology

Article Title: Toll-Interleukin 1 Receptor Domain-Containing Adaptor Protein 180L Single-Nucleotide Polymorphism Is Associated With Susceptibility to Recurrent Pneumococcal Lower Respiratory Tract Infections in Children

doi: 10.3389/fimmu.2018.01780

Figure Lengend Snippet: TLR4 Asp299Gly/Thr399Ile haplotype does not influence IL-6 release in LRTI children. This figure shows scatter dot plots of IL-6 secretion in TLR4 Asp299Gly and Thr399Ile haplotypes co-segregation after (A) LPS or (B) Streptococcus pneumoniae ( Spn ) stimulation. IL-6 secretion is expressed in picograms per milliliter for 10 4 monocytes. Solid line denotes mean. * P < 0.05; ** P < 0.01; *** P < 0.001; and **** P < 0.0001 by Mann–Whitney test. Abbreviations: TLR, toll-like receptor; LRTI, lower respiratory tract infections; IL-6, interleukin-6.

Article Snippet: PE anti-human TLR2 mAb (clone TL2.1, mouse IgG2a κ, RRID:AB_10667887), PE anti-human TLR4 mAb (clone HTA125, mouse IgG2a κ, RRID:AB_1603311), PE isotype control (clone eBM2a, mouse IgG2a κ, RRID:AB_470063), and APC anti-human CD14 mAb (clone 61D3, mouse IgG1, RRID:AB_10669167) were all purchased from eBioscience (THP Medical Products, Switzerland).

Techniques: MANN-WHITNEY

Journal: Frontiers in Immunology

Article Title: Toll-Interleukin 1 Receptor Domain-Containing Adaptor Protein 180L Single-Nucleotide Polymorphism Is Associated With Susceptibility to Recurrent Pneumococcal Lower Respiratory Tract Infections in Children

doi: 10.3389/fimmu.2018.01780

Figure Lengend Snippet: Combined nucleotide polymorphisms in TLR1 or TLR6 and TIRAP are associated with a decreased TLR2 ligand-driven IL-6 release in LRTI children. This figure shows the impact of TIRAP rs8177374, TLR1 rs4833095, TLR1 rs5743618, TLR4 rs4986790, or TLR6 rs5743810 polymorphisms or their combinations on monocyte release of IL-6 following ex vivo whole blood stimulation by LPS, Pam3Cys, or Streptococcus pneumoniae ( Spn ). Values are shown as mean with SEM. Number of children per histogram may differ from those in Table (due to unavailable IL-6 values). (A) Number of patients in each group: wild type (WT) = 20, TIRAP [heterozygous (HT) and homozygous mutant (HM) reported together] = 15, and TLR4 / TIRAP (HT and HM reported together) = 2, TLR4 = 2. (B) WT = 8, TIRAP (HT and HM reported together) = 3, TLR1 rs4833095/ TIRAP (HT and HM reported together) = 12, and TLR1 rs4833095 (HT and HM reported together) = 14. (C) Controls = 6, TIRAP (HT and HM reported together) = 2, TLR1 rs574361 8 / TIRAP (HT and HM reported together) = 13, and TLR1 rs5743618 (HT and HM reported together) = 16. (D) Controls = 13, TIRAP (HT and HM reported together) = 3, TLR6 / TIRAP (HT and HM reported together) = 12, and TLR6 (HT and HM reported together) = 9. Gray histograms represent homozygous wild-type individuals, dashed histograms represent heterozygous, and white histograms represent homozygous mutant. Upper borders of histograms denote mean, and solid horizontal lines denote SEM. IL-6 secretion is expressed in picograms per milliliter for 10 monocytes. * P < 0.05; ** P < 0.01; and *** P < 0.001 by Mann–Whitney test. Abbreviations: TLR, toll-like receptor; WT, wild type; IL-6, interleukin-6.

Article Snippet: PE anti-human TLR2 mAb (clone TL2.1, mouse IgG2a κ, RRID:AB_10667887), PE anti-human TLR4 mAb (clone HTA125, mouse IgG2a κ, RRID:AB_1603311), PE isotype control (clone eBM2a, mouse IgG2a κ, RRID:AB_470063), and APC anti-human CD14 mAb (clone 61D3, mouse IgG1, RRID:AB_10669167) were all purchased from eBioscience (THP Medical Products, Switzerland).

Techniques: Ex Vivo, Mutagenesis, MANN-WHITNEY

Journal: Frontiers in Immunology

Article Title: Toll-Interleukin 1 Receptor Domain-Containing Adaptor Protein 180L Single-Nucleotide Polymorphism Is Associated With Susceptibility to Recurrent Pneumococcal Lower Respiratory Tract Infections in Children

doi: 10.3389/fimmu.2018.01780

Figure Lengend Snippet: Distribution of TLR1, 2, 4, or 6 and TIRAP genotypes in LRTI children.

Article Snippet: PE anti-human TLR2 mAb (clone TL2.1, mouse IgG2a κ, RRID:AB_10667887), PE anti-human TLR4 mAb (clone HTA125, mouse IgG2a κ, RRID:AB_1603311), PE isotype control (clone eBM2a, mouse IgG2a κ, RRID:AB_470063), and APC anti-human CD14 mAb (clone 61D3, mouse IgG1, RRID:AB_10669167) were all purchased from eBioscience (THP Medical Products, Switzerland).

Techniques: Mutagenesis