Journal: Journal of clinical and experimental hematopathology : JCEH

Article Title: Immunohistochemical discrimination of plasmacytoid dendritic cells from myeloid dendritic cells in human pathological tissues.

doi: 10.3960/jslrt.49.23

Figure Lengend Snippet: Fig. 2. Morphological analysis of experimentally induced dendritic cells (DC). (2a) Plasmacytoid dendritic cell (pDC)-precursors from the palatine tonsil exhibiting plasmacytoid appearance ; Giemsa staining. (2b) Mature pDCs induced by treatment with interleukin- 3 and CD40L exhibiting extensively dendritic appearance ; Giemsa staining. (2c-2e) Two-color immunofluorescence analysis of experimentally induced pDCs for S100 calcium binding protein B [S100B ; fluorescein isothiocyanate (FITC)] and fascin [phycoerythrin (PE)] (2c), S100B (FITC) and HLA-DR (PE) (2d), and HLA-DR (FITC) and CD86 (PE) (2e). pDCs are S100B- fascin+ HLA-DR+

Article Snippet: pDCs were generated from purified CD123a+ BDCA2+ pDC-precursors by the modified method described previously.22 Briefly, pDC-precursors were incubated in complete RPMI1640 in the presence of recombinant human interleukin (IL)-3 (50 ng/mL ; R&D Systems, Abingdon, UK) and CD40-ligand (CD40L) recombinant protein (50 ng/ mL ; GenWay Biotech, San Diego, CA) for 3 days at 37°C in 5% CO2 and humidified air.

Techniques: Staining, Immunofluorescence, Binding Assay

Journal: Reproductive Immunology: Open Access

Article Title: NF-kB Regulation in T-cells in Pregnancy is Mediated via Fas/FasL Interactions: The Signal for which is Derived from Exosomes Present in Maternal Plasma

doi: 10.21767/2476-1974.100008

Figure Lengend Snippet: Figure 2: Factors present in maternal plasma can induce apoptosis in Jurkat T-cells. A) Jurkat T-cells were cultured in the presence of Fas activating antibody CH11 (0.625 µg/mL), or recombinant TRAIL (3 ng/mL) and the level of early (DilC1(5)-PI-) and late (DilC1(5)-PI+) apoptosis determined by flow cytometry. Flow charts are representative of 3 separate experiments with similar results. B) Jurkat T-cells were cultured in the presence of 20% (v/v) non-pregnant (NP) or pregnant (P) plasma from individual patients for 72 hrs and the apoptosis determined. Pooled data from 5 individual patients (NP and P15-19) is indicated in the bar graph. C) Jurkat T-cells were pre-incubated for 1 hr in the presence of ZB4 (500 ng/mL) or anti-TRAIL (0.8 ng/mL) and subsequently cultured for 72 hrs in the presence of 20% (v/v) NP or P plasma. Apoptosis was determined by Flow cytometry. The level of early (DilC1(5)-PI-) apoptosis is shown on the graph where n=4 for both NP and P samples. *p<0.05 relative to NP plasma.

Article Snippet: To assess the role of FasL and TRAIL activation in regulating p65 expression and apoptosis cells were incubated in the presence of the Fas activating antibody CH11 (Millipore) or recombinant human TRAIL (ProSci) at concentrations stated in the figure legends.

Techniques: Clinical Proteomics, Cell Culture, Recombinant, Flow Cytometry, Incubation

Journal: Reproductive Immunology: Open Access

Article Title: NF-kB Regulation in T-cells in Pregnancy is Mediated via Fas/FasL Interactions: The Signal for which is Derived from Exosomes Present in Maternal Plasma

doi: 10.21767/2476-1974.100008

Figure Lengend Snippet: Figure 3: Factors present in maternal plasma can induce suppression of p65 in T-cells; A) Jurkat T-cells were cultured in the presence of increasing concentrations of Fas activating antibody CH11 or recombinant TRAIL and the level of p65 determined. Blot is representative of 3 separate experiments with similar results; B) Jurkat T-cells were cultured in the presence of CH11 (0.625 µg/mL) ±500 ng/mL Fas inactivating antibody ZB4 or mIgG control or IgM control (0.625 µg/mL) or recombinant TRAIL (3 ng/mL) ±80 ng/mL anti-TRAIL or mIgG control or BSA (3 ng/mL) and the level of p65 determined. Blot is representative of 3 separate experiments with similar results; C) Jurkat T-cells were cultured in the presence of CH11 (0.625 µg/mL) ±500 ng/mL Fas inactivating antibody ZB4 or mIgG control or IgM control (0.625 µg/mL) and the level of p65 and CD3ζ determined in cell lysates. Blot is representative of 3seperate experiments each showing similar results.

Article Snippet: To assess the role of FasL and TRAIL activation in regulating p65 expression and apoptosis cells were incubated in the presence of the Fas activating antibody CH11 (Millipore) or recombinant human TRAIL (ProSci) at concentrations stated in the figure legends.

Techniques: Clinical Proteomics, Cell Culture, Recombinant, Control