Journal: Nutrients

Article Title: Annona muricata L.-Derived Polysaccharides as a Potential Adjuvant to a Dendritic Cell-Based Vaccine in a Thymoma-Bearing Model

doi: 10.3390/nu12061602

Figure Lengend Snippet: Cytotoxic activity, surface molecule expression, and cytokine production in ALP-stimulated dendritic cells (DCs). DCs were treated with A. muricata L. leaf polysaccharide (ALP) (10, 30, 100, and 200 μg/mL) and LPS (100 ng/mL) for 24 h. ( A ) Cell toxicity was determined using AnnexinV/PI staining (PI + cells; necrosis, AnnexinV + PI + cells; early necrosis, AnnexinV + cells; apoptosis). ( B ) The culture supernatant was used to analyze the production of TNF-α, IL-12p70, and IL-10 using ELISA. ( C ) Intracellular cytokine levels of pro- (TNF-α and IL-12p70) and anti-inflammatory cytokines (IL-10) were assessed in non-, LPS-, and ALP-treated DCs. ( D ) Expression levels of CD80, CD86, MHC-I, and MHC-II in CD11c + -gated cells were detected by FACS. The mean fluorescence intensities of surface molecules in CD11c + cells are shown in each panel. These results are indicated as the mean ± SD ( n = 3 samples) of 3 representative experiments. * p < 0.05, ** p < 0.01, or *** p < 0.001. SD; standard deviation.

Article Snippet: After being stimulated with ALP in the presence of GolgiStop (0.5 μg/mL, BD Bioscience) and GolgiPlug (0.5 μg/mL, BD Bioscience) for 9 h, DCs were stained using Live/Dead Aqua (Live/Dead, BV510, Invitrogen, Carlsbad, CA, USA) and anti-CD11c mAb (PE-Cy7, eBioscience) for 15 min at room temperature.

Techniques: Activity Assay, Expressing, Staining, Enzyme-linked Immunosorbent Assay, Fluorescence, Standard Deviation

Journal: Nutrients

Article Title: Annona muricata L.-Derived Polysaccharides as a Potential Adjuvant to a Dendritic Cell-Based Vaccine in a Thymoma-Bearing Model

doi: 10.3390/nu12061602

Figure Lengend Snippet: Antigen uptake and antigen-presenting ability of ALP-stimulated DCs. ( A ) Cells were stimulated with ALP (200 μg/mL) or LPS (100 ng/mL) for 18 h and cultured with dextran at 37 °C for 30 min. The cells were labeled with anti-CD11c Ab and subjected to FACS analysis to detect dextran uptake. ( B , C ) Non-, LPS-, and ALP-treated cells were then treated with Eα peptide (aa 44–76; 25 μg/mL) or ovalbumin (OVA) (500 μg/mL) for 24 h. After incubating, each group of cells was labeled with anti-CD11c, anti-Y-Ae, or anti-25-D1.16 mAbs for 15 min. Positive controls for antigen presentation, Eα 52–68 (5 μg/mL) or OVA 257–264 (5 μg/mL), were used. Histogram data and bar graphs show the expression of Eα 52–68 /I-Ab ( B ) and OVA 257–264 /H-2Kb ( C ) in the gated CD11c + population. Bar graph data are shown as the mean ± SD ( n = 3 samples) of 3 representative experiments. *** p < 0.001. SD; standard deviation.

Article Snippet: After being stimulated with ALP in the presence of GolgiStop (0.5 μg/mL, BD Bioscience) and GolgiPlug (0.5 μg/mL, BD Bioscience) for 9 h, DCs were stained using Live/Dead Aqua (Live/Dead, BV510, Invitrogen, Carlsbad, CA, USA) and anti-CD11c mAb (PE-Cy7, eBioscience) for 15 min at room temperature.

Techniques: Cell Culture, Labeling, Immunopeptidomics, Expressing, Standard Deviation