Journal: Cell reports

Article Title: Jedi-1/MEGF12-mediated phagocytosis controls the pro-neurogenic properties of microglia in the ventricular-subventricular zone

doi: 10.1016/j.celrep.2023.113423

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: A beveled glass needle was pre-loaded with mineral oil and the recombinant human IL-1 receptor inhibitor (IL-1RA, R&D Systems, Cat #280-RA, resuspended in 0.1% BSA), then positioned in the injecting rig at a 90° angle relative to the horizontal plane of the Petri dish and head mold.

Techniques: Recombinant, Marker, In Situ, RNA Sequencing, Knock-Out, Mutagenesis, Real-time Polymerase Chain Reaction, Software

Journal: The Journal of Experimental Medicine

Article Title: Influence of Polymorphism in the Genes for the Interleukin (IL)-1 Receptor Antagonist and IL-1β on Tuberculosis

doi:

Figure Lengend Snippet: Association between IL-1Ra genotype and the monocyte production of IL-1Ra in response to M . tuberculosis and PPD. The amount of IL-1Ra produced by PBMCs from healthy, PPD-nonreactive donors during a 10-h coculture with either M . tuberculosis (A) or PPD (B) was determined by ELISA. Values were normalized to the number of monocytes present in the culture. The response of IL-1Ra A2 + individuals was higher at all doses of either stimuli.

Article Snippet: After washing (×4), 100 μl of the following biotinylated detection antibodies were added in diluent (0.1% BSA, 0.05% Tween 20 in TBS, pH 7.3): 100 ng/ml anti–human IL-1β antibody or 20 ng/ml anti–human IL-1Ra antibody (both from R & D Systems, Inc.).

Techniques: Produced, Enzyme-linked Immunosorbent Assay

Journal: The Journal of Experimental Medicine

Article Title: Influence of Polymorphism in the Genes for the Interleukin (IL)-1 Receptor Antagonist and IL-1β on Tuberculosis

doi:

Figure Lengend Snippet: Relationship between polymorphism in IL-1Ra and the IL-1Ra/IL-1β ratio. The IL-1β content of the supernates shown in Fig. was also assayed by ELISA. The molar ratio of IL-1Ra/IL-1β was calculated. This ratio is selectively increased in IL-1Ra A2 + individuals in response to PPD and M . tuberculosis , accentuated by the finding that IL-1Ra A2 + subjects tended to produce less IL-1β in response to M . tuberculosis . By comparison, the ratio in response to LPS was not different between IL-1Ra A2 + and A2 − subjects. Response to PPD is data from eight individuals.

Article Snippet: After washing (×4), 100 μl of the following biotinylated detection antibodies were added in diluent (0.1% BSA, 0.05% Tween 20 in TBS, pH 7.3): 100 ng/ml anti–human IL-1β antibody or 20 ng/ml anti–human IL-1Ra antibody (both from R & D Systems, Inc.).

Techniques: Enzyme-linked Immunosorbent Assay, Comparison

Journal: The Journal of Experimental Medicine

Article Title: Influence of Polymorphism in the Genes for the Interleukin (IL)-1 Receptor Antagonist and IL-1β on Tuberculosis

doi:

Figure Lengend Snippet: Mean Fold Induction of the IL-1Rα and IL-1β Genes in Response to M. tuberculosis

Article Snippet: After washing (×4), 100 μl of the following biotinylated detection antibodies were added in diluent (0.1% BSA, 0.05% Tween 20 in TBS, pH 7.3): 100 ng/ml anti–human IL-1β antibody or 20 ng/ml anti–human IL-1Ra antibody (both from R & D Systems, Inc.).

Techniques:

Journal: The Journal of Experimental Medicine

Article Title: Influence of Polymorphism in the Genes for the Interleukin (IL)-1 Receptor Antagonist and IL-1β on Tuberculosis

doi:

Figure Lengend Snippet: (A) Effect of IL-6 and IL-10 on the IL-1Ra/IL-1β ratio. PBMCs were cultured with M . tuberculosis in the presence or absence of rhIL-6 or rhIL-10 over a dose range of 0.1–10 ng/ml. Culture supernates were assayed for IL-1Ra and IL-1β and the molar ratio calculated. IL-10 increased the ratio significantly even at the lowest dose ( P < 0.02), whereas rhIL-6 had no significant effect. (B) Effect of IFN-γ and IL-4 on the IL-1Ra/IL-1β ratio. PBMCs were cultured with M . tuberculosis in the presence or absence of rhIFN-γ or rhIL-4 over a dose range of 0.1–10 ng/ml. Culture supernates were assayed for IL-1Ra and IL-1β and the molar ratio calculated. IL-4 increased the ratio markedly even at the lowest dose ( P < 0.01), whereas higher doses of IFN-γ were required to cause a significant reduction.

Article Snippet: After washing (×4), 100 μl of the following biotinylated detection antibodies were added in diluent (0.1% BSA, 0.05% Tween 20 in TBS, pH 7.3): 100 ng/ml anti–human IL-1β antibody or 20 ng/ml anti–human IL-1Ra antibody (both from R & D Systems, Inc.).

Techniques: Cell Culture

Journal: The Journal of Experimental Medicine

Article Title: Influence of Polymorphism in the Genes for the Interleukin (IL)-1 Receptor Antagonist and IL-1β on Tuberculosis

doi:

Figure Lengend Snippet: Lack of Relationship between IL-1Ra Polymorphism and the Intracellular Growth of M. tuberculosis In Vitro

Article Snippet: After washing (×4), 100 μl of the following biotinylated detection antibodies were added in diluent (0.1% BSA, 0.05% Tween 20 in TBS, pH 7.3): 100 ng/ml anti–human IL-1β antibody or 20 ng/ml anti–human IL-1Ra antibody (both from R & D Systems, Inc.).

Techniques: Infection

Journal: The Journal of Experimental Medicine

Article Title: Influence of Polymorphism in the Genes for the Interleukin (IL)-1 Receptor Antagonist and IL-1β on Tuberculosis

doi:

Figure Lengend Snippet: IL-1Ra and IL-1β Allele and Genotype Frequencies in Tuberculosis Patients and Tuberculin-reactive Healthy Control Subjects

Article Snippet: After washing (×4), 100 μl of the following biotinylated detection antibodies were added in diluent (0.1% BSA, 0.05% Tween 20 in TBS, pH 7.3): 100 ng/ml anti–human IL-1β antibody or 20 ng/ml anti–human IL-1Ra antibody (both from R & D Systems, Inc.).

Techniques: Control

Journal: The Journal of Experimental Medicine

Article Title: Influence of Polymorphism in the Genes for the Interleukin (IL)-1 Receptor Antagonist and IL-1β on Tuberculosis

doi:

Figure Lengend Snippet: Relationship between IL-1Ra/IL-1β Haplotype and Disease Phenotype

Article Snippet: After washing (×4), 100 μl of the following biotinylated detection antibodies were added in diluent (0.1% BSA, 0.05% Tween 20 in TBS, pH 7.3): 100 ng/ml anti–human IL-1β antibody or 20 ng/ml anti–human IL-1Ra antibody (both from R & D Systems, Inc.).

Techniques:

Journal: The Journal of Experimental Medicine

Article Title: Influence of Polymorphism in the Genes for the Interleukin (IL)-1 Receptor Antagonist and IL-1β on Tuberculosis

doi:

Figure Lengend Snippet: Association between IL-1Ra haplotype and the cutaneous response to PPD of M . tuberculosis . The mean diameter of induration in response to one tuberculin unit of PPD in tuberculosis patients of various genotypes is shown. The proportion of IL-1Ra A2 + was highest (60%) in patients with an absent response and lowest in the category 21–30 mm (25%), falling gradually to its overall frequency (41%) as successively higher grades of Mantoux were considered.

Article Snippet: After washing (×4), 100 μl of the following biotinylated detection antibodies were added in diluent (0.1% BSA, 0.05% Tween 20 in TBS, pH 7.3): 100 ng/ml anti–human IL-1β antibody or 20 ng/ml anti–human IL-1Ra antibody (both from R & D Systems, Inc.).

Techniques:

Journal:

Article Title: Computational Design and Application of Endogenous Promoters for Transcriptionally Targeted Gene Therapy for Rheumatoid Arthritis

doi: 10.1038/mt.2009.182

Figure Lengend Snippet: Comparison of therapeutic efficacy using Saa3 versus IL1E/IL6P promoter for disease-regulated Il1rn expression. (a) Promoter activity in transduced synovium of naive and arthritic C57Bl/6 mice. Knee joints were injected with 300 ng p24gag equivalent lentivirus encoding PGK, Saa3, or IL1E/IL6P-luciferase. Seven days after transduction, arthritis was induced by intra-articular injection of 180 µg zymosan A. After 24 hours, luciferase activity was assessed ex vivo. Data are represented as individual relative luciferase activities; horizontal bars indicate the means per group. (b) Efficacy of transcriptionally targeted adenoviral vectors expressing Il1rn. NIH-3T3-5xNF-κB-Luc were transduced at a multiplicity of infection (MOI) of 10 with control vector (del) or adenovirus encoding CMV, Saa3, and IL1E/IL6P-driven Il1rn. After 24 hours, cells were transduced at an MOI of 10 with control vector (del) or Ad5.CMV-Il1b. The day thereafter, IL-1β-induced NF-κB activation was assessed by luciferase assay. Data are represented as relative luciferase activities ± SEM (n = 4), and differences were determined using analysis of variance with Dunnett's post-test. **P < 0.01.

Article Snippet: White high-binding flat bottom 96-well plates (Greiner Bio-One, Alphen a/d Rijn, the Netherlands) were coated with the capture antibody rat anti-murine Il1rn (MAB480; R&D Systems, Minneapolis, MN) at 3 μg/ml in 0.1 mol/l carbonate buffer pH 9.6 and incubated overnight at 4 °C.

Techniques: Comparison, Drug discovery, Expressing, Activity Assay, Injection, Luciferase, Transduction, Ex Vivo, Infection, Control, Plasmid Preparation, Activation Assay

Journal: Journal of Neuroinflammation

Article Title: Anti-inflammatory response following uptake of apoptotic bodies by meningothelial cells

doi: 10.1186/1742-2094-11-35

Figure Lengend Snippet: MECs alter their cytokine and chemokine secretion profile upon uptake of apoptotic bodies. (A) Ben-Men-I cells were treated with PMA, LPS, and left untreated or were incubated with unlabeled apoptotic U-937 cells at a ratio of 1:5 for 24 h. Cell culture supernatants were analyzed for 36 different cytokines and chemokines using an antibody array. The array is organized in five rows (A-E) and 10 columns (1-10) with two spots for each analyte (R = positive control, N = negative control). Please note that spots corresponding to IL-1 receptor antagonist (marked with %) were weak but distinguishable to the eye. Differences in cytokine secretion between control and treatment groups are depicted in qualitative heat maps. (B) Ben-Men-I cells were incubated with unlabeled apoptotic U-937 cells at a ratio of 1:1 or 1:5 for 24 h and the concentration of IL-6 in culture supernatant was measured by ELISA. IL-6 was measured in supernatant of apoptotic cells and subtracted as background from MECs treated with apoptotic cells. Shown is the average of three independent experiments with the error bars representing SD. Statistical significance was assessed using Student’s t -test (* P <0.05). Ben-Men-I cells were treated as in (B) and with concentrations of (C) IL-8, (D) Il-16, (E) CXCL1, (F) MIF (# represents P = 0.067), and (G) IL-1 receptor antagonist were measured by ELISA. Shown is the average of three independent experiments with the error bars representing SD. Statistical significance was analyzed by one-way ANOVA. P values were determined using pair-wise Student’s t -test adjusted according to Holm (# P <0.1; * P <0.05; ** P <0.01; *** P <0.001). Marked are the comparisons between the control (1:0) and treatment groups.

Article Snippet: After 24 h, the culture supernatants were harvested, centrifuged at 1,000 rpm for 5 min, and assayed for IL-6, IL-8 (Orgenium Laboratories), IL-1RA (R&D, DRA00B), IL-16 (RayBiotech, ELH-IL16-001), MIF (RayBiotech, ELH-MIF-001), or CXCL1 (R&D, DRG00) by ELISA.

Techniques: Incubation, Cell Culture, Ab Array, Positive Control, Negative Control, Concentration Assay, Enzyme-linked Immunosorbent Assay