Journal: Journal of Neuroscience

Article Title: System xc Activity and Astrocytes Are Necessary for Interleukin-1 -Mediated Hypoxic Neuronal Injury

doi: 10.1523/jneurosci.2459-07.2007

Figure Lengend Snippet: Figure 2. Role of IL-1RI signaling in the potentiation of hypoxic neuronal injury by IL-1 in vitro. A, B, Mixed cortical cell cultures were treated with 1 ng/ml IL-1 for 20–24 h in the presenceorabsenceofrIL-1ra(10–1000ng/ml;A)oranti-IL-1RI(0.1–100g/ml;B),washed, andthendeprivedofoxygen(5h).Thepercentageoftotalneuronalcelldeathwasdetermined 20–24hlater.Anasteriskindicatesvaluessignificantlygreaterthanhypoxiaalone,whereas# denotes a significant diminution of the IL-1-mediated increase in injury (IL-1) as deter- mined by one-way ANOVA followed by a Student–Newman–Keuls t test. Significance was assessed at p 0.05 (n 3–9 cultures pooled from 2–3 different experiments).

Article Snippet: In experiments using recombinant mouse IL-1ra (rIL-1ra) (R & D Systems) or monoclonal anti-mouse IL-1RI antibody directed against the extracellular domain of mouse IL-1RI (MAB7711; R & D Systems), the respective solutions were prepared with IL-1 (1.5 ng/ml; 1.5 ) and added as described above.

Techniques: In Vitro

Journal: Inflammation and Regeneration

Article Title: Novel artificial nerve transplantation of human iPSC-derived neurite bundles enhanced nerve regeneration after peripheral nerve injury

doi: 10.1186/s41232-024-00319-4

Figure Lengend Snippet: Early-stage tissue evaluation after neurite bundle-derived artificial nerve transplantation. a , b Immunohistochemistry images with anti-NFH and anti-CD31 antibodies. Dotted arrows indicate the proximal side of the intact nerve areas, and solid arrows indicate the NFH- and CD31-positive regeneration tip in the reconstructed tissue. Scale bars = 1 mm. c , d Quantitative comparison of axonal extension distance was indicated by NFH-positive areas and that of vascular elongation distance was demonstrated by CD31-positive areas at 1 and 2 weeks after transplantation. Significant differences between groups elongation distances were observed in 2-week samples only. e , f Iba1 immunohistochemistry image of the proximal part of the regenerated tissue at 2 weeks after transplantation (e). Scale bars = 1 mm. Quantitative analysis of Iba1-positive areas demonstrated that a larger number of macrophages aggregated in the Auto, motor TP and sensory TP (f). ( n = 3). * p < 0.05, ** p < 0.01, N.S. = not significant. Data are represented as the mean ± SEM

Article Snippet: The primary antibodies used in this study were human polyclonal anti-pan-ELAVL (ELAV-like protein 2/3/4) antibody (1:2000, kindly provided from Prof. Robert Darnell, Rockefeller University), rabbit polyclonal anti-TrkA antibody (1:500, kindly provided from Prof. Louis F. Reichardt, UCSF), chicken polyclonal anti-TrkB antibody (1:500, kindly provided from Prof. Louis F. Reichardt, UCSF), goat polyclonal anti-TrkC antibody (AF373, 1:200, R&D systems), rabbit polyclonal anti-parvalbumin antibody (PV-28, 1:1000, Swant), rabbit polyclonal anti-CGRP antibody (BML-CA1134, 1:500, Enzo Life Sciences), Goat polyclonal anti-Choline acetyltransferase antibody (AB144P, 1:200, Chemicon), Mouse monoclonal anti-Islet1 and Islet2 antibody (39.4D5, 1:200, DSHB), rabbit polyclonal anti-Neurofilament heavy polypeptide antibody (ab8135, 1:500, Abcam), goat polyclonal anti-mouse and anti-rat CD31 (AF3628, 1:100, R&D), rabbit polyclonal anti-Iba1 antibody (GtX100042, 1:500, GeneTex), goat anti-GFP antibody (600–101-215, 1:1000, Rockland).

Techniques: Derivative Assay, Transplantation Assay, Immunohistochemistry, Comparison

Journal: Molecular pain

Article Title: Rapid cleavage of IL-1β in DRG neurons produces tissue injury-induced pain hypersensitivity.

doi: 10.1177/17448069241285357

Figure Lengend Snippet: Figure 3. (a) The plantar incision induced a reduction in mechanical threshold against von Frey stimulation. Intrathecal IL-1RA administration significantly inhibited the reduction of the mechanical threshold for 6 h after the treatment. (b) The plantar incision induced a reduction in latency time against thermal stimulation. Intrathecal IL-1RA administration significantly inhibited reduction in the thermal pain latency for 6 h after the treatment. Data were analyzed by repeated measures two-way ANOVA, with Bonferroni’s multiple comparison test comparing pre- and post-treatment time points for each group (#p < .05, ##p < .01, ###p < .001), and Bonferroni’s multiple comparison test comparing both groups at each time point (*p < .05, **p < .01, ***p < .001). The data in the graphs are presented as the mean ± SEM (n = 6 in each group). i.t.: intrathecal administration. Intrathecal administration of IL-1β induced reduction of the mechanical threshold against von Frey stimulation (c) and reduction of latency time against thermal stimulation (d). Data were analyzed by repeated measures two-way ANOVA, with Bonferroni’s multiple comparison test comparing pre- and post-treatment time points for each group (#p < .05, ##p < .01) and Bonferroni’s multiple comparison test comparing both groups at each time point (*p < .05, **p < .01). The data in the graphs are presented as the mean ± SEM (n = 6 in each group).

Article Snippet: Mice were randomly assigned to saline and IL1 RA (500 ng/5 μL, diluted in saline, Recombinant Human IL-1ra/IL-1F3, #280-RA/CF, R&D Systems, Inc. Minneapolis, MN 55413) groups (n = 6 per group).

Techniques: Comparison

Journal:

Article Title: The effect of covalently immobilized rhIL-1ra-ELP fusion protein on the inflammatory profile of LPS-stimulated human monocytes

doi: 10.1016/j.biomaterials.2007.04.010

Figure Lengend Snippet: (a) peptides sequences of IL-1ra-ELP fusion protein (Mw=54,044 Da). The IL-1ra-ELP fusion protein was produced by bacterial expression in E. coli. (b) X-ray crystallography of IL-1ra with the lysines highlighted in magenta. (c) An image of six-well tissue culture polystyrene plates with a 5 nm Cr adhesion layer and a 45 nm Au layer evaporated yielding optically transparent gold-coated slides.

Article Snippet: The bioactivity of IL-1ra-ELP was benchmarked against commercially available human recombinant IL-1ra (rhIL-1ra) (R&D Systems, Minneapolis, MN) by IL-1ra inhibition of IL-1 induced proliferation of RPMI 1788 cells [ 28 , 29 ] using ATP production as the cell viability assay.

Techniques: Produced, Expressing

Journal:

Article Title: The effect of covalently immobilized rhIL-1ra-ELP fusion protein on the inflammatory profile of LPS-stimulated human monocytes

doi: 10.1016/j.biomaterials.2007.04.010

Figure Lengend Snippet: (a) Bioactivity of IL-1ra-ELP fusion protein and commercial rhIL-1ra using the inhibition assay of IL-1β stimulated RPMI 1788 cells. The percentage of inhibited proliferation is plotted as a function of excess molar IL-1ra added in 10−2–104. The amount of inhibitor, IL-1ra, for inhibition of stimulatory effect of IL-1β is determined based on the 4-parameter sigmoidal fitting curve. EC50 of IL-1ra-ELP = 211.21 ± 0.31 pM (R2= 0.92) and EC50 Commercial rhIL-1ra = 20.21 ± 0.85 pM (R2= 0.97) Data presented as mean ± SE (n=4). Inset: standard curve of IL-1β induction of RPMI 1788 proliferation.

Article Snippet: The bioactivity of IL-1ra-ELP was benchmarked against commercially available human recombinant IL-1ra (rhIL-1ra) (R&D Systems, Minneapolis, MN) by IL-1ra inhibition of IL-1 induced proliferation of RPMI 1788 cells [ 28 , 29 ] using ATP production as the cell viability assay.

Techniques: Inhibition

Journal:

Article Title: The effect of covalently immobilized rhIL-1ra-ELP fusion protein on the inflammatory profile of LPS-stimulated human monocytes

doi: 10.1016/j.biomaterials.2007.04.010

Figure Lengend Snippet: The amount of IL-1ra-ELP detected from SAM(a), Imm IL-1ra(b) and Sol IL-1ra(c) in the absence of THP-1 monocytes and secreted IL-1ra from the Imm IL-1ra(d) and Sol IL-1ra(e) in the presence of THP-1 monocytes over time course as detected using the LINCO plex assay. No IL-1ra on the Imm IL-1ra in the absence of THP-1 monocytes was detected over the time course tested up to 5 days showing the immobilized IL-1ra-ELP is stable on the SAM surfaces. Severed IL-1ra-ELP is the IL-1ra-ELP detached from the surface possibly due to unstable chemical coupling or enzymatic action from the monoctyes. Secreted IL-1ra is the IL-1ra secreted from THP-1 monocytes responding to the external stimuli. Added IL-1ra-ELP is the soluble IL-1ra-ELP directly added into the media. * The concentration of IL-1ra at 1 hr was close to the maximun detection limit of LINCOplex assay. Sol IL-1ra w/o THP-1 was tested up to 3 days and Sol IL-1ra with THP-1 was tested up to 5 days. N.D. not detected.

Article Snippet: The bioactivity of IL-1ra-ELP was benchmarked against commercially available human recombinant IL-1ra (rhIL-1ra) (R&D Systems, Minneapolis, MN) by IL-1ra inhibition of IL-1 induced proliferation of RPMI 1788 cells [ 28 , 29 ] using ATP production as the cell viability assay.

Techniques: Plex Assay, Concentration Assay

Journal:

Article Title: The effect of covalently immobilized rhIL-1ra-ELP fusion protein on the inflammatory profile of LPS-stimulated human monocytes

doi: 10.1016/j.biomaterials.2007.04.010

Figure Lengend Snippet: Schematic illustration of human THP-1 monocytes interfering with the four experimental conditions used in the cell culture experiments. i.e. (a) MHA SAMs (SAM), (b) SAMs with immobilized BSA, (c) SAMs with immobilized IL-1ra-ELP (Imm IL1-ra), and (d) MHA SAMs with soluble IL-1ra (Sol IL-1ra), each of which were seeded with either presence or absence of 1 μg/ml of LPS. LPS added to the culture well at the time when the THP-1 monocytes were seeded.

Article Snippet: The bioactivity of IL-1ra-ELP was benchmarked against commercially available human recombinant IL-1ra (rhIL-1ra) (R&D Systems, Minneapolis, MN) by IL-1ra inhibition of IL-1 induced proliferation of RPMI 1788 cells [ 28 , 29 ] using ATP production as the cell viability assay.

Techniques: Cell Culture

Journal:

Article Title: The effect of covalently immobilized rhIL-1ra-ELP fusion protein on the inflammatory profile of LPS-stimulated human monocytes

doi: 10.1016/j.biomaterials.2007.04.010

Figure Lengend Snippet: (a–d) phase contrast images of THP-1 monocytes to show the effect of immobilized and soluble IL-1ra-ELP fusion protein on THP-1 differentiation. Cell images cultured on (a) MHA SAM without LPS stimulation, (b) immobilized IL-1ra-ELP on the MHA SAM surface with LPS stimulation, (c) soluble IL-1ra-ELP (1 μg/ml) was directly added into the media with LPS stimulation and (d) MHA SAM with LPS stimulation, were collected 72 h after seeding. (a–c) all possessed a rounded, undifferentiated morphology. Cells circled in (d) indicate a flattened, differentiated morphology of THP-1 cells. Magnification 200X.

Article Snippet: The bioactivity of IL-1ra-ELP was benchmarked against commercially available human recombinant IL-1ra (rhIL-1ra) (R&D Systems, Minneapolis, MN) by IL-1ra inhibition of IL-1 induced proliferation of RPMI 1788 cells [ 28 , 29 ] using ATP production as the cell viability assay.

Techniques: Cell Culture

Journal:

Article Title: The effect of covalently immobilized rhIL-1ra-ELP fusion protein on the inflammatory profile of LPS-stimulated human monocytes

doi: 10.1016/j.biomaterials.2007.04.010

Figure Lengend Snippet: Qualitative comparison of the relative expression levels of the 10 cytokines assayed at 72hrs under the four experimental conditions with and without LPS stimulation. * The level of IL-1β was tested using ELISA assay because of detection limit of Lincoplex. N.D. is non-detectable, i.e. zero. “+” , “ ++ ” and “ +++ ” mean relatively l ow, moderate and high, respectively. “ − ” means detected less than 50 pg/ml for IL-1β, IL-4, IL-6, IL-8, TNF-α, IFN-γ, MCP-1 and IL-1ra and 100 pg/ml for MIP-1α and VEGF

Article Snippet: The bioactivity of IL-1ra-ELP was benchmarked against commercially available human recombinant IL-1ra (rhIL-1ra) (R&D Systems, Minneapolis, MN) by IL-1ra inhibition of IL-1 induced proliferation of RPMI 1788 cells [ 28 , 29 ] using ATP production as the cell viability assay.

Techniques: Comparison, Expressing, Enzyme-linked Immunosorbent Assay, Control

Journal:

Article Title: The effect of covalently immobilized rhIL-1ra-ELP fusion protein on the inflammatory profile of LPS-stimulated human monocytes

doi: 10.1016/j.biomaterials.2007.04.010

Figure Lengend Snippet: Bar graphs of expression levels of IL-1β, TNF-α, IL-6, IL-8, MIP-1α, MCP-1, VEGF and IL-1ra at 1, 6, 24, 48 and 72 hrs from LPS-stimulated monocytes on MHA SAMs, SAMs with immobilized IL-1ra-ELP, and MHA SAMs with soluble IL-1ra-ELP as detected using the LINCOplex assay. IL-1β expression under the same conditions as measured by ELISA due to detection limit of the LINCOplex. * Significant relative to the SAM; p<0.05

Article Snippet: The bioactivity of IL-1ra-ELP was benchmarked against commercially available human recombinant IL-1ra (rhIL-1ra) (R&D Systems, Minneapolis, MN) by IL-1ra inhibition of IL-1 induced proliferation of RPMI 1788 cells [ 28 , 29 ] using ATP production as the cell viability assay.

Techniques: Expressing, Enzyme-linked Immunosorbent Assay