Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Human anti-IgG1 hinge autoantibodies reconstitute the effector functions of proteolytically inactivated IgGs.

doi: 10.4049/jimmunol.181.5.3183

Figure Lengend Snippet: FIGURE 2. Binding of IgG3 autoantibodies to four different intact mAb IgG1s as well as to Fab and F(ab)2 generated from them with various proteases. A, Binding of serum IgG3 from 20 individual healthy donors. Symbols represent the average ELISA signal obtained for each serum sam- ple in three replicate analyses, and the mean value for the 20 samples is indicated by the horizontal line. The specific fragments of IgGs with the sites of proteolytic cleavage are indicated on the x-axis. B, Detection of IgG3 autoantibody binding to the same group of IgGs and fragments from pooled human IVIg at 2 mg/ml. Bar heights correspond to the mean SEM from three tests. C, Depiction of the sequence of the IgG1 H chain

Article Snippet: Isotype standards of IgG1, IgG2, IgG3, IgG4, IgM, IgE, and IgD were from myeloma serum and were obtained from Athens Research & Technology.

Techniques: Binding Assay, Generated, Enzyme-linked Immunosorbent Assay, Sequencing

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: NPJ Vaccines

Article Title: LmCen −/− based vaccine is protective against canine visceral leishmaniasis following three natural exposures in Tunisia

doi: 10.1038/s41541-025-01070-8

Figure Lengend Snippet: a Protocol for analysis of the Immunogenicity of Lm Cen −/− . Three groups of dogs were included in this step: dogs receiving 10 6 or 10 7 live Lm Cen −/− parasites and as negative control dogs receiving 100 µl of medium. Immunization was done by intradermal injection of 100 µl of vaccine in the ear of dog. Boost with vaccine was done at the same conditions as the first immunization at an interval of 2 months. b– d Histograms represent the mean of total IgG anti-SLA at different kinetics. Vertical bars indicate the standard deviation (SD). Total IgG titers are measured using SLA from L.major . Statistical analyses are done using GraphPad Prism, * p < 0.05: difference is statistically significant.

Article Snippet: The plates were washed six times, and horseradish peroxidase-conjugated rabbit anti-dog total immunoglobulin G (IgG) (Sigma-Aldrich), goat anti-dog IgG1 or sheep anti-dog IgG2a (Bethyl Laboratories) was added for 1 h at 37 °C, at dilutions of 1:10,000.

Techniques: Immunopeptidomics, Negative Control, Injection, Standard Deviation

Journal: NPJ Vaccines

Article Title: LmCen −/− based vaccine is protective against canine visceral leishmaniasis following three natural exposures in Tunisia

doi: 10.1038/s41541-025-01070-8

Figure Lengend Snippet: IgG anti- SGH and anti-SLA are measured in sera from dogs immunized with Lm Cen −/− at a dose of 10 6 and those receiving 100 µl of PBS (Negative control). Titers of IgG anti-SGH measured in dogs before ( a ) or at return from the field after successive expositions 2019 ( b ), 2020 ( c ) and 2021 ( d ). Titers of IgG anti-SLA before ( e ) and after the exposure in the endemic area ( f–h ) measured using SLA from L.infantum . Histograms represent the mean of IgG anti-SGH or anti-SLA for each group. Bars represent the standard deviation for each group. Pie charts represent number of dog positive and/or negative for IgG anti-SLA and IgG anti-SGH at return from the field in 2019 ( i – l ), 2020 ( j – m ) and 2021 ( k–n ). Cutoff of IgG anti-SLA is fixed at OD = 0.23; cutoff for IgG anti-SGH fixed at OD = 0.08.

Article Snippet: The plates were washed six times, and horseradish peroxidase-conjugated rabbit anti-dog total immunoglobulin G (IgG) (Sigma-Aldrich), goat anti-dog IgG1 or sheep anti-dog IgG2a (Bethyl Laboratories) was added for 1 h at 37 °C, at dilutions of 1:10,000.

Techniques: Negative Control, Standard Deviation

Journal: Journal of veterinary diagnostic investigation : official publication of the American Association of Veterinary Laboratory Diagnosticians, Inc

Article Title: Diagnostic gap in Bovine viral diarrhea virus serology during the periparturient period in cattle.

doi: 10.1177/1040638713501172

Figure Lengend Snippet: Figure 2. Bovine immunoglobulin G (IgG) concentrations in serum of 5 cows antepartum (a.p.), at birth (p.), and postpartum (p.p.). For the detailed sampling time points, see Figure 1 (serum neutralization test [SNT]). The total immunoglobulin (IgG) (A), IgG1 (B), and IgG2 (C) concentrations were determined by enzyme-linked immunosorbent assay (ELISA).i Dashed lines highlight animals that showed a diagnostic gap in Bovine viral diarrhea virus (BVDV) antibody ELISAa around parturition. The IgG1-to-IgG2 ratio (D) was calculated for the antepartum serum samples.

Article Snippet: Bovine IgG Quantitation ELISA Set, Bethyl Laboratories Inc., Montgomery, TX. j. Sheep anti-bovine IgG1 antibody, HRP conjugated, Bethyl Laboratories Inc., Montgomery, TX. k. Sheep anti-bovine IgG2 antibody, HRP conjugated, Bethyl Laboratories Inc., Montgomery, TX. l. Purified bovine IgG1, Bethyl Laboratories Inc., Montgomery, TX. m. Purified bovine IgG2, Bethyl Laboratories Inc., Montgomery, TX.

Techniques: Sampling, Neutralization, Enzyme-linked Immunosorbent Assay, Diagnostic Assay, Virus

Journal: Journal of veterinary diagnostic investigation : official publication of the American Association of Veterinary Laboratory Diagnosticians, Inc

Article Title: Diagnostic gap in Bovine viral diarrhea virus serology during the periparturient period in cattle.

doi: 10.1177/1040638713501172

Figure Lengend Snippet: Figure 3. Immunoglobulin G (IgG) class-specific Bovine viral diarrhea virus (BVDV) antibody enzyme-linked immunosorbent assay (ELISA) results (relative optical density [OD] values) in serum of 5 cows antepartum (a.p.), at birth (p.), and postpartum (p.p.). For the detailed sampling time points, see Figure 1 (serum neutralization test [SNT]). The total anti-BVDV IgG (A), anti-BVDV IgG1 (B), and anti-BVDV IgG2 (C) OD values were determined by an adapted in-house ELISA. Dashed lines highlight animals that showed a diagnostic gap in BVDV antibody ELISAa around parturition. The anti-BVDV IgG1-to-IgG2 ratio (D) was calculated for the antepartum serum samples.

Article Snippet: Bovine IgG Quantitation ELISA Set, Bethyl Laboratories Inc., Montgomery, TX. j. Sheep anti-bovine IgG1 antibody, HRP conjugated, Bethyl Laboratories Inc., Montgomery, TX. k. Sheep anti-bovine IgG2 antibody, HRP conjugated, Bethyl Laboratories Inc., Montgomery, TX. l. Purified bovine IgG1, Bethyl Laboratories Inc., Montgomery, TX. m. Purified bovine IgG2, Bethyl Laboratories Inc., Montgomery, TX.

Techniques: Virus, Enzyme-linked Immunosorbent Assay, Sampling, Neutralization, Diagnostic Assay

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