Journal: eLife

Article Title: Microglia TREM2R47H Alzheimer-linked variant enhances excitatory transmission and reduces LTP via increased TNF-α levels

doi: 10.7554/elife.57513

Figure Lengend Snippet: Figure 7. LTP is impaired in RH mutant rats, and the impairment could be rescued by application of anti-TNF-a antibody. (A) The input-output slope is significantly increased in Trem2R47H/R47H rats [two-way ANOVA, stimulation intensity x genotype interaction F(6, 144)=6.745, p<0.0001****; post-hoc Sidak’s multiple comparisons test: 1.2 mV p=0.0175*; 1.4 mV: p=0.0021**; 1.6 mV: p=0.0001***] Representative traces of fEPSPs in response to increasing stimulus from 0.4 to 1.6 mA are shown on the top. (B) LTP is impaired in Trem2R47H/R47H rats. Average traces of the baseline and the last 5mins of LTP are shown on top. (C) Plot of fEPSP slope change of the last 10 min of LTP in B (unpaired t test, p<0.0001****). (D) The increase of input- out slope in Trem2R47H/R47H rats is reversed by application of anti-TNF-a [ANOVA for repeated measures F(18, 300)=6.579, p<0.0001****; post-hoc Tukey’s multiple comparisons test: 0.8 mV RH/RH + Isotype vs. w/w + anti-TNF-a p=0.0479*; 1.0 mV RH/RH + Isotype vs. RH/RH + anti-TNF-a p=0.0061**, RH/RH + Isotype vs. w/w + anti-TNFa p=0.0072**, RH/RH + Isotype vs. w/w + Isotype p=0.0113*; 1.2 mV RH/RH + Isotype vs. RH/RH + anti-TNFa p=0.0009***, RH/RH + Isotype vs. w/w + anti-TNF-a p=0.0009***, RH/RH + Isotype vs. w/w + Isotype p=0.0029**; 1.4 mV RH/RH + Isotype vs. RH/RH + anti-TNF-a p<0.0001****, RH/RH + Isotype vs. w/w + anti-TNF-a p<0.0001****, RH/RH + Isotype vs. w/w + Isotype p=P < 0.0001****; 1.6 mV RH/RH + Isotype vs. RH/RH + anti-TNF-a p<0.0001****, RH/RH + Isotype vs. w/w + anti-TNF-a P p<0.0001****, RH/RH + Isotype vs. w/w + Isotype p<0.0001****]. Representative fEPSP traces are shown on top. (E) The impaired LTP is restored by application of anti-TNF-a antibody. The average traces of the baseline and the last 5mins of LTP are shown on top. (F) Plot of fEPSP slope change of the last 10 min of LTP in E [one-way ANOVA, F(3,

Article Snippet: Aging Cell 18: e13033 Rat App allele with humanize Ab region Genetic reagent (Rattus Norvegicus) Trem2R47H Tambini and D’Adamio, 2020 Sci Rep 10: 4122 Rat Trem2 allele with R47H mutation Commercial assay or kit V-PLEX Plus Ab Peptide Panel 1 Meso Scale Discovery Cat# K15200G Used following manufacturer’s recommendations Commercial assay or kit V-PLEX Proinflammatory Panel 2 Meso Scale Discovery Cat# K15059D Used following manufacturer’s recommendations Commercial assay or kit CD11b/c (Microglia) Micro-Beads, rat antibody Cat# 130-105-634 Miltenyi Biotec RRID:AB_2783886 Used following manufacturer’s recommendations Commercial assay or kit Adult Brain Dissociation Kit Miltenyi Biotec Cat# 130-107-677 Used following manufacturer’s recommendations Commercial assay or kit RNeasy RNA Isolation kit Qiagen Cat# 74106 Used following manufacturer’s recommendations Commercial assay or kit High-Capacity cDNA RT kit Thermo Cat# 4368814) Used following manufacturer’s recommendations Commercial assay or kit TaqMan Fast Advanced Mix Thermo Cat# 4444556 Used following manufacturer’s recommendations Commercial assay or kit Gapdh RealTime PCR Thermo Rn01775763_g1 Used following manufacturer’s recommendations Commercial assay or kit Treml1 RealTime PCR Thermo Rn01511908_g1 Used following manufacturer’s recommendations Antibody Polyclonal Goat IgG anti-Rat TNFa Cat# AF-510-NA R and D Systems RRID:AB_354511 10 ng/ml in ACSF Antibody Polyclonal Goat IgG. antibody Cat# AB-108-C R and D Systems RRID:AB_354267 10 ng/ml in ACSF Software, algorithm LinRegPCR software hartfaalcentrum.nl Software, algorithm pCLAMP10 software Molecular Devices, Software, algorithm Image Lab software Biorad RRID:SCR_014210 Software, algorithm GraphPad Prism RRID:SCR_002798 Ren et al. eLife 2020;9:e57513.

Techniques: Mutagenesis

Journal: Clinical and Developmental Immunology

Article Title: Inhibition of Arterial Allograft Intimal Hyperplasia Using Recipient Dendritic Cells Pretreated with B7 Antisense Peptide

doi: 10.1155/2012/892687

Figure Lengend Snippet: BM-DCs detected by inverted microscope and scanning electron microscope. Progenitor cells were propagated in the presence of rmGM-CSF. After TNF- α and donor antigen addition, the cells show significant differentiation with typical DC appearance under inverted microscope (a) and scanning electron microscope (b) and (c).

Article Snippet: In the sixth day, the donor antigen and recombinant mouse TNF- α (rmTNF- α , 50 μ g/L; R&D Systems, Minneapolis, MN, USA) were added to the medium.

Techniques: Inverted Microscopy, Microscopy

Journal: Journal of biomedical materials research. Part A

Article Title: Effects of hyaluronic acid conjugation on anti-TNF-α inhibition of inflammation in burns

doi: 10.1002/jbm.a.34829

Figure Lengend Snippet: Binding affinity experiments. Association and dissociation curves for (a.) (anti-TNF-α)-HA and (b.) anti-TNF-α. Vertical line separates association, on the left from dissociation. c. Comparison of initial loading of biotinylated antibody and conjugate onto streptavidin tips. Greater thickness is observed for the conjugate possibly due to interactions with HA chains.

Article Snippet: The experimental parameters consisted of the following dipping sequence: PBS 1 min (baseline), anti-TNF-α or (anti-TNF-α)-HA (10 μg/mL antibody) 20 min (loading), biocytin (10 μg/mL) 3 min (quench), PBS 5 min (wash), PBS 5 min (baseline), recombinant rat TNF-α (R&D systems Inc, Minneapolis, MN) 20 min (association), PBS 60 min (dissociation).

Techniques: Binding Assay

Journal: Journal of biomedical materials research. Part A

Article Title: Effects of hyaluronic acid conjugation on anti-TNF-α inhibition of inflammation in burns

doi: 10.1002/jbm.a.34829

Figure Lengend Snippet: Masson’s trichrome images of burn edges. Granulation tissue started to appear four days after eschar removal in the deep dermal region, with the most robust granulation tissue in the (anti-TNF-α)-HA treated sites. HA, anti-TNF-α and anti-TNF-α + HA treatments shows slower granulation tissue formation, while saline treated wound sites display the slowest granulation tissue formation. On day 4, newly forming eschar (#) is observed in most of the wound sites, and on day 7, necrosis has grown significantly particularly in saline and anti-TNF-α treated burn sites. Adjacent tissue (*) appears healthiest day 7 in the (anti-TNF-α)-HA treated sites. Scale bar = 1mm.

Article Snippet: The experimental parameters consisted of the following dipping sequence: PBS 1 min (baseline), anti-TNF-α or (anti-TNF-α)-HA (10 μg/mL antibody) 20 min (loading), biocytin (10 μg/mL) 3 min (quench), PBS 5 min (wash), PBS 5 min (baseline), recombinant rat TNF-α (R&D systems Inc, Minneapolis, MN) 20 min (association), PBS 60 min (dissociation).

Techniques:

Journal: Journal of biomedical materials research. Part A

Article Title: Effects of hyaluronic acid conjugation on anti-TNF-α inhibition of inflammation in burns

doi: 10.1002/jbm.a.34829

Figure Lengend Snippet: H & E staining. Day 7 images of the burn at the interface of wound edge and newly formed eschar shows densely populated multinucleated inflammatory immune cells apparent in saline and anti-TNF-α treatments. These immune cells, indicated with an arrow, are less densely populated in HA and anti-TNF-α + HA, with the fewest and most sparse found with (anti-TNF-α)-HA treatments. Scale bar = 200 μm.

Article Snippet: The experimental parameters consisted of the following dipping sequence: PBS 1 min (baseline), anti-TNF-α or (anti-TNF-α)-HA (10 μg/mL antibody) 20 min (loading), biocytin (10 μg/mL) 3 min (quench), PBS 5 min (wash), PBS 5 min (baseline), recombinant rat TNF-α (R&D systems Inc, Minneapolis, MN) 20 min (association), PBS 60 min (dissociation).

Techniques: Staining

Journal: Journal of biomedical materials research. Part A

Article Title: Effects of hyaluronic acid conjugation on anti-TNF-α inhibition of inflammation in burns

doi: 10.1002/jbm.a.34829

Figure Lengend Snippet: Vimentin immunostaining was used to assess viable tissue, by measuring thickness of nonviable tissue. Trichrome images were referenced if it was not immediately clear where the viable tissue ended. By day 7, (anti-TNF-α)-HA has decreased the thickness of necrosis significantly. (Anti-TNF-α)-HA treated burn sites have significantly thinner nonviable zones than saline and anti-TNF-α treated sites and denser vimentin staining in the viable region. Thickness of nonviable tissue in anti-TNF-α treated sites is not significantly different from saline treatments, but trends towards a slight reduction in nonviable tissue. *: p<0.05; error bars = mean +/− SD

Article Snippet: The experimental parameters consisted of the following dipping sequence: PBS 1 min (baseline), anti-TNF-α or (anti-TNF-α)-HA (10 μg/mL antibody) 20 min (loading), biocytin (10 μg/mL) 3 min (quench), PBS 5 min (wash), PBS 5 min (baseline), recombinant rat TNF-α (R&D systems Inc, Minneapolis, MN) 20 min (association), PBS 60 min (dissociation).

Techniques: Immunostaining, Staining

Journal: Journal of biomedical materials research. Part A

Article Title: Effects of hyaluronic acid conjugation on anti-TNF-α inhibition of inflammation in burns

doi: 10.1002/jbm.a.34829

Figure Lengend Snippet: Macrophage infiltration counts. On day 1, macrophage count appears to be most affected by (anti-TNF-α)-HA, and slightly by anti-TNF-α treatment. Anti-TNF-α treatments appeared to decrease macrophage counts the most by days 4 and 7. (Anti-TNF-α)-HA treatment attenuated slightly by day 7, but was still significantly decreased compared to saline control (* p<.001, bars p<.05; error bars are mean +/− SD).

Article Snippet: The experimental parameters consisted of the following dipping sequence: PBS 1 min (baseline), anti-TNF-α or (anti-TNF-α)-HA (10 μg/mL antibody) 20 min (loading), biocytin (10 μg/mL) 3 min (quench), PBS 5 min (wash), PBS 5 min (baseline), recombinant rat TNF-α (R&D systems Inc, Minneapolis, MN) 20 min (association), PBS 60 min (dissociation).

Techniques:

Journal: Journal of biomedical materials research. Part A

Article Title: Effects of hyaluronic acid conjugation on anti-TNF-α inhibition of inflammation in burns

doi: 10.1002/jbm.a.34829

Figure Lengend Snippet: IL-1β concentration in extracted burn tissue. IL-1β concentration peaks at day 1, and attenuates on day 4 in saline and (anti-TNF-α)-HA treated wound sites, however IL-1β levels are significantly lower than those in saline treated sites throughout the experimental period (* p<0.01 day 4, and p<0.05 day 7). Anti-TNF-α treatment exhibited the opposite response to saline and (anti-TNF-α)-HA, inhibiting IL-1β day 1 compared to saline, but then steadily loses effect by days 4 and 7. Error bars = mean +/− SD.

Article Snippet: The experimental parameters consisted of the following dipping sequence: PBS 1 min (baseline), anti-TNF-α or (anti-TNF-α)-HA (10 μg/mL antibody) 20 min (loading), biocytin (10 μg/mL) 3 min (quench), PBS 5 min (wash), PBS 5 min (baseline), recombinant rat TNF-α (R&D systems Inc, Minneapolis, MN) 20 min (association), PBS 60 min (dissociation).

Techniques: Concentration Assay