Journal: Acta Neuropathologica Communications

Article Title: Brain injury drives optic glioma formation through neuron-glia signaling

doi: 10.1186/s40478-024-01735-w

Figure Lengend Snippet: a Following optic nerve crush (ON-CR) at 6 weeks of age, Nf1 flox/flox ; hGFAP-Cre mice at 12 weeks of age have increased numbers of CD3 + cells in their optic nerves relative to those receiving a sham operation ( n = 6); however, there was no change in b Ccl4 mRNA expression by qPCR ( n = 3). c Immediately after ON-CR, Nf1 flox/flox ; hGFAP-Cre mice received intraperitoneal injections of anti-CD3 (αCD3) antibodies every other day for 6 weeks. Control mice were injected with anti-IgG antibodies. Optic nerves were analyzed at 12 weeks of age. αCD3 antibody treatment (150 µg) reduced d CD3 + cell content ( n = 5) and e proliferation (%Ki67 + cells; n = 5). f αCD3 antibody treatment following ON-CR does not change Ccl5 RNA expression by qPCR compared to IgG-treated controls (150 µg; n = 3). Increased Ccl5 RNA expression in the optic nerves (qPCR) was observed 7 days after ON-CR in g Nf1 flox/flox ; hGFAP-Cre ( n = 3), h athymic ( Foxn1 nu/nu ; n = 4), and i Rag1 −/− mice ( n = 4) compared to sham controls. Scale bars: a , d , e 50 µm. Two-tailed Student’s t test (ns, not significant)

Article Snippet: Four-week-old Nf1 OPG mice were treated with 10 mg/kg NFκB inhibitor (NFκB-IN; Caffeic acid phenethyl ester, Fisher Scientific, 274310), 275 mg/kg PLX3397-containing or control chow pellets (Free Base), 1 mg/ml anti-IL-1β neutralizing antibodies (R&D System 1060-DE-100), anti-IgG2a control antibodies (R&D Systems), or memantine hydrochloride (20 mg/kg) by intraperitoneal injection.

Techniques: Expressing, Control, Injection, RNA Expression, Two Tailed Test

Journal: Acta Neuropathologica Communications

Article Title: Brain injury drives optic glioma formation through neuron-glia signaling

doi: 10.1186/s40478-024-01735-w

Figure Lengend Snippet: a Top 3 identified pathways enriched in the optic nerves of Nf1 flox/flox ; hGFAP-Cre mice after optic nerve crush (ON-CR) relative to sham controls following filtering of differentially expressed transcripts from bulk RNA sequencing. b Increased IL-1β RNA expression (qPCR) is observed in the optic nerves of 12-week-old Nf1 flox/flox ; hGFAP-Cre mice following ON-CR at 6 weeks of age relative to sham controls ( n = 4). c Representative IL-1β immunostaining in the optic nerves of Nf1 flox/flox ; hGFAP-Cre mice following ON-CR relative to sham control mice. d RNAscope (in situ RNA hybridization) demonstrates that oligodendrocytes (Olig2 + cells) express IL-1β. e Immunostaining reveals that the majority of the Olig2 + cells co-label with CC1, but not with NG2. f Increasing IL-1β concentrations (0–75 ng/ml) increases microglia Ccl5 protein expression in vitro. g IL-1β-induced microglial Ccl5 production is attenuated by 10 mg/kg NFκB inhibitor (NFκB-IN; Caffeic acid phenethyl ester) treatment in vitro ( n = 4). h Anti-IL1β neutralizing antibody treatment (1 mg/ml) of Nf1 flox/flox ; hGFAP-Cre mice immediately after ON-CR at 6 weeks of age results in decreased i optic nerve proliferation (%Ki67 + cells; n = 5) and j Ccl5 mRNA expression (qPCR; n = 3) when analyzed at 12 weeks of age compared to IgG-treated (1 mg/ml) controls. Data are presented as the means ± SEM. Scale bars: c , d , e , i 50 µm. b , c , e , i , j . Two-tailed Student’s t test; f , g One-way ANOVA with Bonferroni post-test correction

Article Snippet: Four-week-old Nf1 OPG mice were treated with 10 mg/kg NFκB inhibitor (NFκB-IN; Caffeic acid phenethyl ester, Fisher Scientific, 274310), 275 mg/kg PLX3397-containing or control chow pellets (Free Base), 1 mg/ml anti-IL-1β neutralizing antibodies (R&D System 1060-DE-100), anti-IgG2a control antibodies (R&D Systems), or memantine hydrochloride (20 mg/kg) by intraperitoneal injection.

Techniques: RNA Sequencing, RNA Expression, Immunostaining, Control, RNAscope, In Situ, Hybridization, Expressing, In Vitro, Two Tailed Test

Journal: Acta Neuropathologica Communications

Article Title: Brain injury drives optic glioma formation through neuron-glia signaling

doi: 10.1186/s40478-024-01735-w

Figure Lengend Snippet: a Traumatic brain injury (TBI) in Nf1 flox/flox ; hGFAP-Cre mice at 6 weeks of age results in increased b optic nerve volume ( n = 6) and c proliferation (%Ki67 + cells; n = 7) relative to sham treated mice when analyzed at 12 weeks of age, as well as increased TAMs (%Iba1 + cells) and CD3 + T cell content. Increased d Ccl5 RNA expression (qPCR) and e glutamate levels are observed in the optic nerves of Nf1 flox/flox ; hGFAP-Cre mice ( n = 4) 7 days after TBI compared to sham controls. f αIL-1β neutralizing antibody (1 mg/ml) and h memantine treatment (20 mg/kg) both decrease proliferation (%Ki67 + cells; n = 5) and g , i Ccl5 expression ( n = 3) following TBI relative to their respective control mice (IgG and vehicle treatment groups). Data are presented as the means ± SEM. Scale bar: b 100 μm; c , f , i 50 μm. Two-tailed Student’s t test

Article Snippet: Four-week-old Nf1 OPG mice were treated with 10 mg/kg NFκB inhibitor (NFκB-IN; Caffeic acid phenethyl ester, Fisher Scientific, 274310), 275 mg/kg PLX3397-containing or control chow pellets (Free Base), 1 mg/ml anti-IL-1β neutralizing antibodies (R&D System 1060-DE-100), anti-IgG2a control antibodies (R&D Systems), or memantine hydrochloride (20 mg/kg) by intraperitoneal injection.

Techniques: RNA Expression, Expressing, Control, Two Tailed Test

Journal: Infection and Immunity

Article Title: Role of T Cells and Gamma Interferon during Induction of Hypersensitivity to Lipopolysaccharide by Toxic Shock Syndrome Toxin 1 in Mice

doi: 10.1128/iai.69.3.1256-1264.2001

Figure Lengend Snippet: FIG. 4. Effects of Cs (A) or anti-IFN-g MAb (B) on LPS-induced serum TNF-a activity in BALB/c-AnNCr mice primed with TSST-1. Groups of four mice were pretreated with Cs (40 mg/kg, i.p.) or anti-IFN-g MAb (750 mg/mouse, i.p.) at 4 or 2 h, respectively, prior to injection of TSST-1. Mice were then injected i.p. with 200 mg of TSST-1 (dotted bars) per kg or PBS (clear bars). Control mice received equal volumes of PBS instead of Cs, rat IgG2a instead of anti-IFN-g MAb, or PBS instead of TSST-1. All mice were injected with LPS (400 mg/kg, i.p.) 12 h later. TNF-a activity in serum was measured 2 h postinjection of LPS. Solutions of Cs or MAb were treated with 2 mg of polymyxin B per ml to remove contaminating LPS. The percent reductions in circulating TNF-a activity caused by Cs or anti-IFN-g MAb are shown.

Article Snippet: Neutralizing rat monoclonal antibody (MAb) to mouse IFN-g, rat immunoglobulin G2a (IgG2a) isotype control MAb, and murine recombinant TNF-a were obtained from R&D Systems (Minneapolis, Minn.).

Techniques: Activity Assay, Injection, Control