Journal: Science advances

Article Title: FasL microgels induce immune acceptance of islet allografts in nonhuman primates.

doi: 10.1126/sciadv.abm9881

Figure Lengend Snippet: Fig. 1. Local presentation of SA-FasL via synthetic microgels for islet transplantation. (A) Synthetic biotin-PEG microgels, generated by microfluidic polymerization, capture, and present SA-FasL. SA-FasL–presenting microgels and islets are immobilized on the surface of the omentum of NHPs using autologous plasma and thrombin, where they induce immune acceptance by potentially eliminating T effector cells and generating T regulatory cells. (B) Schema describing donor and diabetic MHC mis- matched recipient NHPs and treatment protocol. SA-FasL–presenting microgels with transient rapamycin (RAPA) monotherapy (SA-FasL Microgels, n = 4); control micro- gels with transient rapamycin monotherapy (Microgels, n = 3).

Article Snippet: Detection of SA-FasL protein and anti-SA and anti-FasL antibody The presence and quantity of SAFasL protein in the plasma of SA FasL microgel–treated NHPs was assessed using a rat FasL sandwich ELISA kit from RayBiotech, with a sensitivity of 90 pg/ml.

Techniques: Transplantation Assay, Generated, Clinical Proteomics, Control

Journal: Mediators of Inflammation

Article Title: Amphiregulin Regulates Phagocytosis-Induced Cell Death in Monocytes via EGFR and Matrix Metalloproteinases

doi: 10.1155/2018/4310419

Figure Lengend Snippet: Neonatal monocytes display elevated pro-AREG surface expression and increased release of AREG in response to E. coli infection. PBMO and CBMO were incubated with E. coli for 1 h, extracellular bacteria were removed, and cells were cultivated for 24 h in total. (a) Immunocytological staining shows AREG protein expression (green) in monocytes. Texas Red-X phalloidin was applied to visualize F-actin in the cytoskeleton (red). (b) Pro-AREG surface expression in uninfected and E. coli -infected monocytes was quantified by using flow cytometry ( n = 5). Representative dot plots show gating strategy and cutoff value for AREG expression. (c) sAREG levels in the supernatant of monocytes were quantified by using ELISA ( n = 5). Data are shown as means + SD. Statistical analysis was performed using one-way ANOVA with Bonferroni's multiple comparisons test (ns: not significant, ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗∗ p < 0.001).

Article Snippet: For detection of FasL, Human FasL PicoKine ELISA Kit (Boster Biological Technology, Pleasanton, USA) was used according to the manufacturer's instructions.

Techniques: Expressing, Infection, Incubation, Bacteria, Staining, Flow Cytometry, Enzyme-linked Immunosorbent Assay

Journal: Mediators of Inflammation

Article Title: Amphiregulin Regulates Phagocytosis-Induced Cell Death in Monocytes via EGFR and Matrix Metalloproteinases

doi: 10.1155/2018/4310419

Figure Lengend Snippet: AREG mediates increased shedding of memFasL through engagement with EGFR. PBMO and CBMO were infected as described in . AREG stimulation and EGFR inhibitor treatment was started 1 h prior to infection and was maintained during cultivation. (a) memFasL surface expression in uninfected and E. coli -infected monocytes was analyzed in response to AREG stimulation and neutralization of EGFR by using flow cytometry ( n = 5). AREG stimulation significantly decreased FasL levels in infected PBMO but not CBMO. Neutralization of EGFR resulted in a complete rescue of infection-induced memFasL presentation. Representative dot plots show gating strategy and cutoff value. (b) sFasL levels in the supernatant of uninfected and E. coli -infected monocytes were quantified in response to AREG stimulation and neutralization of EGFR by using ELISA ( n = 3). AREG significantly increased sFasL levels in infected PBMO but not CBMO, while neutralization of EGFR abolished the effect. Data are shown as means + SD. Statistical significance was analyzed using two-way ANOVA with Bonferroni's multiple comparisons test ( ∗∗ p < 0.01, ∗∗∗ p < 0.005, and ∗∗∗∗ p < 0.001).

Article Snippet: For detection of FasL, Human FasL PicoKine ELISA Kit (Boster Biological Technology, Pleasanton, USA) was used according to the manufacturer's instructions.

Techniques: Infection, Expressing, Neutralization, Flow Cytometry, Enzyme-linked Immunosorbent Assay

Journal: Mediators of Inflammation

Article Title: Amphiregulin Regulates Phagocytosis-Induced Cell Death in Monocytes via EGFR and Matrix Metalloproteinases

doi: 10.1155/2018/4310419

Figure Lengend Snippet: EGFR-dependent gelatinase activation mediates increased shedding of memFasL. PBMO and CBMO were infected as described in . Stimulation with AREG and gelatinase inhibition by CHX treatment was started 1 h prior to infection and was maintained during cultivation. (a) memFasL surface expression in uninfected and E. coli -infected monocytes was analyzed in response to AREG stimulation and gelatinase inhibition by using flow cytometry ( n = 4). Gelatinase inhibition prevented the decrease in memFasL presentation on monocytes triggered by AREG. (b) sFasL levels in the supernatant of uninfected and E. coli -infected monocytes were quantified in response to AREG stimulation and CHX treatment by using ELISA ( n = 3). Gelatinase inhibition was found to prevent the increase in sFasL levels triggered by AREG. Data are shown as means + SD. Statistical significance was analyzed using two-way ANOVA with Bonferroni's multiple comparisons test ( ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗∗ p < 0.001).

Article Snippet: For detection of FasL, Human FasL PicoKine ELISA Kit (Boster Biological Technology, Pleasanton, USA) was used according to the manufacturer's instructions.

Techniques: Activation Assay, Infection, Inhibition, Expressing, Flow Cytometry, Enzyme-linked Immunosorbent Assay

Journal: Renal Failure

Article Title: Cisplatin-induced oxidative stress stimulates renal Fas ligand shedding

doi: 10.1080/0886022X.2018.1456938

Figure Lengend Snippet: Cisplatin-induced oxidative stress increases urinary sFasL in mice. Bar graphs summarizing the levels of sFasL in: (A) Plasma, and (B) Urine of Captisol (vehicle control)-, cisplatin (15 mg/kg; single IP injection)-, and TEMPOL (100 mg/kg; IP for 4 days) + cisplatin-treated mice. * p < .05 vs. Captisol; # p < .05 vs. cisplatin; n = 6 each.

Article Snippet: Plasma and urinary concentrations of FasL were quantified using the mouse sFasL PicoKine ELISA kit (Boster Biological Technology, Pleasanton, CA).

Techniques: Clinical Proteomics, Control, Injection

Journal: Scientific Reports

Article Title: Glucocorticoids impair oocyte developmental potential by triggering apoptosis of ovarian cells via activating the Fas system

doi: 10.1038/srep24036

Figure Lengend Snippet: Cortisol injection increases the level of glucorticoids (GC) in blood and ovary. Within the ovary, the GC elevation activates the Fas/FasL system and decreases levels of growth factors and the E2/P4 ratio by down-regulating the expression of GR. The activation of Fas/FasL and the decrease in growth factors and E2/P4 ratio work together to induce apoptosis of ovarian cells, leading to a decline in growth factor levels and E2/P4 ratio but an increase in FasL in the follicular fluid (FF). The increase in FasL and the decrease in growth factor levels and the E2/P4 ratio in FF cause apoptosis in both CCs and oocytes, impairing oocyte developmental potential. Furthermore, the apoptotic CCs also produce FasL that affects oocytes by acting on their Fas receptors.

Article Snippet: ELISA for CRH and FasL was performed using CRH Elisa kits (E03P0031) and FasL Elisa kits (E03F0051) purchased from Shanghai BlueGene Biological Technology Co., Ltd.

Techniques: Injection, Expressing, Activation Assay