Journal: International Journal of Molecular Sciences

Article Title: TNF-α Affects Signature Cytokines of Th1 and Th17 T Cell Subsets through Differential Actions on TNFR1 and TNFR2

doi: 10.3390/ijms23169306

Figure Lengend Snippet: TNF-α does not impact the levels of TNFR1 and TNFR2 on Th1 and Th17 cells. Purified CD4 + T cell subsets were stimulated with 1 µg/mL of TNF-α for 24 h and stained, for flow cytometry, with anti-human TNFR1 or TNFR2 mAbs. ( A ) Representative histograms showing the expression levels of TNFR1 and TNFR2 on Th1 and Th17 cells at baseline conditions and upon TNF-α stimulus. Solid black line histograms, isotype control; black histograms, Th1 cells; grey histograms, Th17 cells; dashed line histograms, TNF-α-treated Th1 cells; long dashed line histograms, TNF-α-treated Th17 cells. The levels of TNFR1 ( B ) and TNFR2 ( C ) were measured on purified CD4 + T cell subpopulations obtained from two to four healthy controls. Bars represent the mean values ± SD. Statistical analyses were carried out with Kruskal–Wallis followed by Dunn’s multiple comparison tests. ** p < 0.01.

Article Snippet: Cells were then stained with a mouse anti-human TNFR1-APC mAb (clone 16803) (R&D Systems, Minneapolis, MN, USA) and a rat anti-human TNFR2-PE mAb (clone hTNFR-M1) (BD Biosciences) for 30 min at 4 °C in the dark.

Techniques: Purification, Staining, Flow Cytometry, Expressing, Control, Comparison

Journal: International Journal of Molecular Sciences

Article Title: TNF-α Affects Signature Cytokines of Th1 and Th17 T Cell Subsets through Differential Actions on TNFR1 and TNFR2

doi: 10.3390/ijms23169306

Figure Lengend Snippet: Effect of TNFR1 or TNFR2 blockade on the production of IFN-γ and IL-17 by Th1 and Th17 cells. Purified CD4 + T lymphocyte subpopulations were incubated with neutralizing antibodies to TNFR1 or TNFR2 for 1 h prior to a 4-day stimulus with TNF-α (1 µg/mL). Intracellular staining of IFN-γ and IL-17 was assessed by flow cytometry. ( A ) Representative dot plots of Th1 and Th17 cells treated with anti-TNFR1 or anti-TNFR2 in the presence or absence of TNF-α. An isotype control was used to discard non-specific effects of the neutralizing antibodies. The fold increase in the percentages of IFN-γ ( B , D ) or IL-17 ( C , E ) producers was measured on Th1 and Th17 cells obtained from two to six healthy donors. Bars represent the mean values ± SD. Data were normalized against cells that did not receive treatment with TNF-α or TNF-α plus TNFRs blocking mAbs. For statistical analysis, Kruskal–Wallis and Dunn’s multiple comparison tests were performed. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Cells were then stained with a mouse anti-human TNFR1-APC mAb (clone 16803) (R&D Systems, Minneapolis, MN, USA) and a rat anti-human TNFR2-PE mAb (clone hTNFR-M1) (BD Biosciences) for 30 min at 4 °C in the dark.

Techniques: Purification, Incubation, Staining, Flow Cytometry, Control, Blocking Assay, Comparison

Journal: International Journal of Molecular Sciences

Article Title: TNF-α Affects Signature Cytokines of Th1 and Th17 T Cell Subsets through Differential Actions on TNFR1 and TNFR2

doi: 10.3390/ijms23169306

Figure Lengend Snippet: Expression of TNFR1 and TNFR2 on Th1 and Th17 cells present in the peripheral blood of rheumatoid arthritis (RA) patients treated with adalimumab. Cell staining for flow cytometry analysis was performed on PBMC samples from healthy controls ( n = 9) and RA patients ( n = 10) before (PRE) and after (POST) treatment with adalimumab. The levels of TNFR1 ( A ) and TNFR2 ( C ) are expressed in MFI values. The frequency of TNFR1 ( B ) and TNFR2 ( D )-expressing lymphocytes are also shown. Each symbol represents data for one individual. Mean values ± SD are indicated. Significance was assessed with non-parametric Kruskal–Wallis test followed by Dunn’s multiple comparison test (for MFI data) or parametric one-way ANOVA plus Tukey’s post-test (for lymphocyte frequencies data). * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: Cells were then stained with a mouse anti-human TNFR1-APC mAb (clone 16803) (R&D Systems, Minneapolis, MN, USA) and a rat anti-human TNFR2-PE mAb (clone hTNFR-M1) (BD Biosciences) for 30 min at 4 °C in the dark.

Techniques: Expressing, Staining, Flow Cytometry, Comparison

Journal: Cell Reports

Article Title: Selective functional antibody transfer into the breastmilk after SARS-CoV-2 infection

doi: 10.1016/j.celrep.2021.109959

Figure Lengend Snippet:

Article Snippet: Mouse Anti-Human IgG2-Fc PE , Southern Biotech , CAT # 9060-09.

Techniques: Virus, Luciferase, Recombinant, Produced, Generated, Software, Luminex

Journal: PLOS ONE

Article Title: A mechanistically novel peptide agonist of the IL-7 receptor that addresses limitations of IL-7 cytokine therapy

doi: 10.1371/journal.pone.0286834

Figure Lengend Snippet: (A, B) Determination of IC 50 values in a competition ELISA for compound binding to human and cyno IL-7Rα and γc. The assay format is IL-7Rα-(His) 6 -tagged ECD or Fc-γc ECD immobilized on 96-well plates. Tracers are C-terminal biotinylated forms of the reference IL-7Rα and γc peptide ligands, each pre-complexed with Neutravidin-HRP (NA-HRP). (C, D) Label-free measurement of the equilibrium dissociation constant (K D ) for peptide binding to IL-7Rα and γc extracellular domains with peptides was performed by biolayer interferometry as detailed in methods. Red lines depict 1:1 binding fit from Gator™ software. Blue lines represent individual BLI sensogram data taken every 0.1 seconds. Inset table displays comparison of ELISA data and BLI kinetic data.

Article Snippet: Biotinylated mouse anti-human IgG2 Fc (Southern Biotech Cat# 9070–08) was used as the capture antibody, and the rabbit polyclonal antibody against MDK1188, followed by a mouse anti-rabbit IgG Fc HRP (Genscript Cat# A01856-200) as the detection antibody.

Techniques: Enzyme-linked Immunosorbent Assay, Binding Assay, Software, Comparison

Journal: PLoS ONE

Article Title: CD4 and MHCII phenotypic variability of peripheral blood monocytes in dogs

doi: 10.1371/journal.pone.0219214

Figure Lengend Snippet: Antibodies (Ab) used in the study.

Article Snippet: 4 , CD11b , mouse anti-dog , CA16.3E10 , IgG1 , unconjugated , AbD Serotec.

Techniques:

Journal: Vaccines

Article Title: Elucidating the Onset of Cross-Protective Immunity after Intranasal Vaccination with the Attenuated African Swine Fever Vaccine Candidate BA71ΔCD2

doi: 10.3390/vaccines12050517

Figure Lengend Snippet: The presence of ASFV-specific IgG before the challenge, but not IgM, is associated with the control of infection. The induction of ASFV-specific IgM ( A ) and IgG ( B ) antibodies was measured for each animal by an ELISA at the indicated time points. Black dotted lines represent the vaccination and challenge time points. ( C ) Analysis of ASFV-specific IgG1 and IgG2 isotypes in sera (1/100 dilution) at 12 days postvaccination. ( D ) ASFV-specific IgG antibody titers were compared between unvaccinated animals and animals vaccinated 3 days prior to the challenge. The mean and standard deviation of the average IgG levels from each group and time point are represented. Statistical significance was assessed by a paired t -test (ns p > 0.05; * p ≤ 0.05; *** p ≤ 0.001).

Article Snippet: Peroxidase-conjugated rabbit anti-pig IgG (1/20.000 dilution; Sigma-Aldrich, Saint Louis, MO, USA), goat anti-pig IgM (1/20.000 dilution; BioRad, Hercules, CA, USA), mouse anti-pig IgG1 (1/1000; BioRad), mouse anti-pig IgG2 (1/1000; BioRad) and peroxidase-conjugated goat anti-mouse IgG (A, G, M) (1/2500; Sigma-Aldrich) were used as secondary or detection antibodies for the presence of ASF-specific immunoglobulins.

Techniques: Control, Infection, Enzyme-linked Immunosorbent Assay, Standard Deviation

Journal: Cell reports

Article Title: Cooperativity mediated by rationally selected combinations of human monoclonal antibodies targeting the henipavirus receptor binding protein

doi: 10.1016/j.celrep.2021.109628

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Mouse Anti-Human IgG2 Fc-AP , Southern Biotech , Cat# 9070-04.

Techniques: Recombinant, Virus, Transfection, Expressing, Sterility, Antibody Labeling, Plasmid Preparation, Software, Chromatography, Microscopy