Journal: Pain medicine (Malden, Mass.)

Article Title: Neonatal bee venom exposure induces sensory modality-specific enhancement of nociceptive response in adult rats.

doi: 10.1111/pme.12296

Figure Lengend Snippet: Figure 1 A flowchart illustrating the experimental design.

Article Snippet: After being blocked in 5% normal goat serum in PBS containing 0.3% Triton X-100 for 30 minutes, sections were incubated overnight at 4°C with primary antibodies to NGF (1:200; rabbit anti-rat, Santa Cruz Biotechnology, Santa Cruz, CA, USA), TrkA receptor (1:200; rabbit antirat, Santa Cruz), BDNF (1:200; rabbit anti-rat, Santa Cruz), TrkB receptor (1:200; rabbit anti-rat, Santa Cruz), IL-1β (1:200; rabbit anti-rat, Santa Cruz), or COX-2 (1:200; goat anti-rat, Santa Cruz).

Techniques:

Journal: Pain medicine (Malden, Mass.)

Article Title: Neonatal bee venom exposure induces sensory modality-specific enhancement of nociceptive response in adult rats.

doi: 10.1111/pme.12296

Figure Lengend Snippet: Figure 8 Effects of neonatal inflammation on the spinal expression of IL-1β and COX-2 in adult rats. Compared with adult rats without neonatal inflammation, rats receiving the first BV injection on P21 and beyond showed up-regulated expression of IL-1β in the spinal cord dorsal horn. Scale bar: 200 μm. IOD = integral optical density. Sal-BV = rats receiving saline injection neonatally and then exposed to BV as adults. BV-BV = rats receiving BV neonatally and then again exposed to BV as adults. * P < 0.05, as compared with the Sal-BV group.

Article Snippet: After being blocked in 5% normal goat serum in PBS containing 0.3% Triton X-100 for 30 minutes, sections were incubated overnight at 4°C with primary antibodies to NGF (1:200; rabbit anti-rat, Santa Cruz Biotechnology, Santa Cruz, CA, USA), TrkA receptor (1:200; rabbit antirat, Santa Cruz), BDNF (1:200; rabbit anti-rat, Santa Cruz), TrkB receptor (1:200; rabbit anti-rat, Santa Cruz), IL-1β (1:200; rabbit anti-rat, Santa Cruz), or COX-2 (1:200; goat anti-rat, Santa Cruz).

Techniques: Expressing, Injection, Saline

Journal: PLoS Pathogens

Article Title: IL-33 receptor ST2 regulates the cognitive impairments associated with experimental cerebral malaria

doi: 10.1371/journal.ppat.1006322

Figure Lengend Snippet: (A-B) Mixed glial cell cultures from newborn WT (A) and ST2 -/- (B) mice were stimulated with rmIL-33 (10 or 100ng/mL) or LPS (10μg/mL), as a positive control for 24 hrs, or unstimulated, then fixed and used for immunofluorescence staining of IL-1β with microglia marker CD11b. Arrows indicate colocalization. These images are representative of 3 independent experiments. Scale bar 100μm. (C) Cell percentages of CD11b + IL-1β + cells from A and B. These results are expressed as mean ± SEM, n = 3 mice per group in 2 independent experiments with 5 fields analyzed per slide. Kruskal-Wallis was applied, followed by Dunn’s comparison test (*p≤0 . 05 , **p≤0 . 01) .

Article Snippet: The cells were incubated overnight with different primary antibodies raised in rat anti-CD11b (Bio-Rad MCA711G; 1:200), goat anti-IL-1β (Sigma I3767; 1:500), or rabbit anti-GFAP, rabbit anti-OLIG2, goat anti-IL-33 as above.

Techniques: Positive Control, Immunofluorescence, Staining, Marker

Journal: PLoS Pathogens

Article Title: IL-33 receptor ST2 regulates the cognitive impairments associated with experimental cerebral malaria

doi: 10.1371/journal.ppat.1006322

Figure Lengend Snippet: (A) WT mixed glial cell cultures from newborn mice were stimulated with IL-1β (30ng/mL) or LPS (10μg/mL), as a positive control, for 24 hrs, then fixed and immunostained for OLIG2 and IL-33. These images are representative of 2 independent experiments. Arrows indicate colocalization of IL-33 with OLIG2. Scale bar 100μm. (B) Percentage of OLIG2 + cells expressing IL-33 from A. These results are expressed as mean ± SEM, n = 3 mice per group in 2 independent experiments with 5 fields analyzed per slide. Kruskal-Wallis was applied, followed by Dunn’s comparison test (**p≤0.01) .

Article Snippet: The cells were incubated overnight with different primary antibodies raised in rat anti-CD11b (Bio-Rad MCA711G; 1:200), goat anti-IL-1β (Sigma I3767; 1:500), or rabbit anti-GFAP, rabbit anti-OLIG2, goat anti-IL-33 as above.

Techniques: Positive Control, Expressing

Journal: PLoS Pathogens

Article Title: IL-33 receptor ST2 regulates the cognitive impairments associated with experimental cerebral malaria

doi: 10.1371/journal.ppat.1006322

Figure Lengend Snippet: In the CNS IL-33 is expressed by endothelial cells, astrocytes and prominently by oligodendrocytes. After Pb A-infection, IL-33 induces IL-1β release by microglia, contributing to the high levels of IL-1β, TNF-α and IFN-γ production seen at an early stage of infection. The neuroinflammatory response induced by IL-33/ST2 pathway alters neuroblast proliferation, associated with neurogenesis impairment and cognitive defects. In addition, IL-1-β induced by microglia, in turns stimulates oligodendrocytes to produce high levels of IL-33, leading to an amplification loop. IL-33 overexpression by oligodendrocytes induced an inflammatory context which could affect the myelin structure, as reported after Pb A-infection in WT mice , and this could be responsible for some cognitive impairments. Furthermore, IL-33/ST2 pathway orchestrates CNS glial cell response at an early stage of Pb A-infection, promoting CXCL9 and CXCL10 production to recruit, in a second phase the ECM pathogenic CD8 + T cells in cerebral vessels.

Article Snippet: The cells were incubated overnight with different primary antibodies raised in rat anti-CD11b (Bio-Rad MCA711G; 1:200), goat anti-IL-1β (Sigma I3767; 1:500), or rabbit anti-GFAP, rabbit anti-OLIG2, goat anti-IL-33 as above.

Techniques: Infection, Amplification, Over Expression

Journal: Innate immunity

Article Title: Myeloid-derived NF-κB negative regulation of PU.1 and cEBPβ-driven pro-inflammatory cytokine production restrains LPS-induced Shock

doi: 10.1177/1753425916681444

Figure Lengend Snippet: (A). U-LPS (E. coli K12)–challenged survival of p65Δmye and littermate WT mice. Serum cytokine levels of B) IL-1β, C) IL-6, and D) TNF-α at indicated time points (hours) in WT and p65Δmye mice following one i.p. injection with U-LPS (E. coli K12) (40 mg/kg). Data represent the mean ± SEM of 2 independent experiments with (A). n = 7–11 mice per group and (B – D). n = 3–11 mice per group. (*p < 0.05) between groups.

Article Snippet: The following antibodies were used: rabbit anti-mouse caspase-1 p10 (M20; Santa Cruz; Santa Cruz, CA) and goat anti-mouse IL-1β/IL-1F2 (R&D Systems; Minneapolis, Minn) followed by goat anti-rabbit (Calbiochem; Darmstadt, Germany) or sheep anti-goat (Jackson ImmunoResearch Laboratories; West Grove, PA) peroxidase-conjugated antibodies.

Techniques: Injection

Journal: Innate immunity

Article Title: Myeloid-derived NF-κB negative regulation of PU.1 and cEBPβ-driven pro-inflammatory cytokine production restrains LPS-induced Shock

doi: 10.1177/1753425916681444

Figure Lengend Snippet: A and B, Flow cytometry analyses of F4/80, CD11b, and Ly6C expression in WT and p65Δmye BMDMs, respectively. Panel labels of a-d represent indicated gatings. C, Pro-inflammatory cytokine IL-1β, IL-6, and TNF-α secretion from BMDMs pretreated for 0, 3, or 6 h with 100 ng/mL U-LPS (E. coli K12) followed by 1 h of 2 mM ATP stimulation. D, Viability of BMDMs determined by MTT assay. Cells were treated with 0, 1, 10 100, or 1000 ng/mL U-LPS (E. coli K12) for 24 h prior to MTT assay. Data represent the mean ± SEM from n = 2–3 separate experiments. Significant differences indicated (*p < 0.05; **p < 0.01; ***p < 0.005) between groups. N.D., not detectable.

Article Snippet: The following antibodies were used: rabbit anti-mouse caspase-1 p10 (M20; Santa Cruz; Santa Cruz, CA) and goat anti-mouse IL-1β/IL-1F2 (R&D Systems; Minneapolis, Minn) followed by goat anti-rabbit (Calbiochem; Darmstadt, Germany) or sheep anti-goat (Jackson ImmunoResearch Laboratories; West Grove, PA) peroxidase-conjugated antibodies.

Techniques: Flow Cytometry, Expressing, MTT Assay

Journal: Innate immunity

Article Title: Myeloid-derived NF-κB negative regulation of PU.1 and cEBPβ-driven pro-inflammatory cytokine production restrains LPS-induced Shock

doi: 10.1177/1753425916681444

Figure Lengend Snippet: A, Pro-inflammatory cytokine IL-1β, IL-6, and TNF-α secretion; B, Il1b and caspase-1 gene expression analyzed by quantitative real-time PCR; C, Western blot of pro-IL-1β expression in unstimulated peritoneal inflammatory MΦs. D, Western blot of pro-IL-1β, IL-1β, pro-caspase-1, and caspase-1 expression of peritoneal MΦs pretreated for 6 h with 0, 10, or 100 ng/mL U-LPS (E. coli K12) followed by a 1-h stimulation with 2 mM ATP. Data represent the mean ± SEM of 2 independent experiments with n = 2–3 mice per group. Cytokine expression is normalized to the expression of the reference gene (hprt). Significant differences (*p < 0.05; **p < 0.01; ***p < 0.005) between groups. N.D., not detectable.

Article Snippet: The following antibodies were used: rabbit anti-mouse caspase-1 p10 (M20; Santa Cruz; Santa Cruz, CA) and goat anti-mouse IL-1β/IL-1F2 (R&D Systems; Minneapolis, Minn) followed by goat anti-rabbit (Calbiochem; Darmstadt, Germany) or sheep anti-goat (Jackson ImmunoResearch Laboratories; West Grove, PA) peroxidase-conjugated antibodies.

Techniques: Expressing, Real-time Polymerase Chain Reaction, Western Blot