Journal: International Journal of Nanomedicine

Article Title: In vitro toxicity studies of biodegradable, polyelectrolyte nanocapsules

doi: 10.2147/IJN.S169120

Figure Lengend Snippet: Analysis of inflammatory response to nanocapsules. Notes: ( A ) Viability of mouse macrophage cell line P388D1 and endothelial MBE cells. The cells were incubated for 48 hours in growth medium with 1.5 mM NaCl or nanocapsules (~2×10 11 /mL). ( B ) Analysis of NO production by iNOS. Nitrite levels were measured by Griess reaction after 24 hours incubation of the cells with 1) nanocapsules or NaCl (NT), 2) LPS or cytokines and 3) nanocapsules and LPS or cytokines. ( C ) Flow cytometry analysis of VCAM-1 expression on MBE cells. MBE cells were incubated for 24 hours with nanocapsules or with TNF and then stained with anti-VCAM-1 antibody. * P <0.05 Abbreviations: LPS, lipopolysaccharide; MFI, mean fluorescence intensity; NC5-PEG, five-layer PEGylated nanocapsules; NC6, six-layer nanocapsules; NO, nitric oxide; PEG, polyethylene glycol; TNF, tumor necrosis factor; VCAM, vascular cell adhesion molecule.

Article Snippet: Human hepatoma-derived HepG2 cell line (ATCC ® HB-8065) and mouse macrophage cell line P388D1 (ATCC ® CCL-46) were obtained from American Type Culture Collection.

Techniques: Incubation, Flow Cytometry, Expressing, Staining, Fluorescence

Journal: International Journal of Nanomedicine

Article Title: In vitro toxicity studies of biodegradable, polyelectrolyte nanocapsules

doi: 10.2147/IJN.S169120

Figure Lengend Snippet: Quantification of DNA breaks after exposure of PBMC, P388D1 and HepG2 cells to nanocapsules – comet assay. The cells were cultured for 2, 24 or 48 hours in media containing 1.5 mM NaCl or nanocapsules (~2×10 11 /mL) or etoposide. * P <0.05. Abbreviations: NC5-PEG, five-layer PEGylated nanocapsules; NC6, six-layer nanocapsules; PBMC, peripheral blood mononuclear cell; PEG, polyethylene glycol.

Article Snippet: Human hepatoma-derived HepG2 cell line (ATCC ® HB-8065) and mouse macrophage cell line P388D1 (ATCC ® CCL-46) were obtained from American Type Culture Collection.

Techniques: Single Cell Gel Electrophoresis, Cell Culture

Journal:

Article Title: Modulation of Dengue Virus Infection in Human Cells by Alpha, Beta, and Gamma Interferons

doi:

Figure Lengend Snippet: Dose response of cytokine pretreatment on DV2 infection of HepG2 cells. HepG2 cells were preincubated with increasing concentrations of IFN-β or -γ or TNF-α for 24 h and then exposed to DV2 (strain 16681) at an MOI of 2; 72 h later, cells (6 × 105) were detached and subjected to immunofluorescent flow cytometry after incubation with FITC-labeled anti-DV2 antibody. The results are expressed as the percentage of cells that express the E protein of DV2; 48% of cells that were infected without cytokine pretreatment (negative media control) were positive by flow cytometry. One representative experiment of two is shown.

Article Snippet: Murine hybridomas against human DV antigens (3H5-1, anti-DV2; 5D4-11, anti-DV3; and 2H2-9, anti-DV) or flavivirus antigens (4G2) were obtained from the ATCC and grown in Dulbecco's modified Eagle medium (Gibco BRL) supplemented with 10% FBS, 2 mM l -glutamine, 1 mM sodium pyruvate, penicillin G (100 U/ml), and streptomycin (100 μg/ml) at 37°C in 5% CO 2 .

Techniques: Infection, Flow Cytometry, Incubation, Labeling

Journal:

Article Title: Modulation of Dengue Virus Infection in Human Cells by Alpha, Beta, and Gamma Interferons

doi:

Figure Lengend Snippet: Time course of the effect of IFN pretreatment on DV2 infection of HFF cells. HFF cells were exposed to DV2 (strain 16681) and incubated for 72 h. Cells (2 × 105) were processed for flow cytometry (A), and supernatants were harvested for plaque assays (B) using BHK-21 cells. In each case, cells were exposed to medium IFN-γ (10 ng/ml), IFN-β (10 ng/ml), or IFN-β+γ (10 ng/ml) either 24 or 4 h before (pre) or after (post) incubation with virus. The flow cytometric data are presented as the percentage of cells that express the E protein of DV2, and the plaque assay data are expressed as the number of PFU per milliliter. One representative experiment of three is shown.

Article Snippet: Murine hybridomas against human DV antigens (3H5-1, anti-DV2; 5D4-11, anti-DV3; and 2H2-9, anti-DV) or flavivirus antigens (4G2) were obtained from the ATCC and grown in Dulbecco's modified Eagle medium (Gibco BRL) supplemented with 10% FBS, 2 mM l -glutamine, 1 mM sodium pyruvate, penicillin G (100 U/ml), and streptomycin (100 μg/ml) at 37°C in 5% CO 2 .

Techniques: Infection, Incubation, Flow Cytometry, Plaque Assay

Journal:

Article Title: Modulation of Dengue Virus Infection in Human Cells by Alpha, Beta, and Gamma Interferons

doi:

Figure Lengend Snippet: Time course of the effect of IFN pretreatment on DV2 infection by low-passage viral isolates. HepG2 cells were exposed to DV2 (Thai strains C0477 and K0049; Nicaraguan strain N9622) at an MOI of 2 and incubated for 72 h. In each case, cells were exposed to medium or combinations of IFN-β and -γ (10 ng/ml each) as described for Fig. ​Fig.2.2. One representative experiment of two is shown.

Article Snippet: Murine hybridomas against human DV antigens (3H5-1, anti-DV2; 5D4-11, anti-DV3; and 2H2-9, anti-DV) or flavivirus antigens (4G2) were obtained from the ATCC and grown in Dulbecco's modified Eagle medium (Gibco BRL) supplemented with 10% FBS, 2 mM l -glutamine, 1 mM sodium pyruvate, penicillin G (100 U/ml), and streptomycin (100 μg/ml) at 37°C in 5% CO 2 .

Techniques: Infection, Incubation

Journal:

Article Title: Modulation of Dengue Virus Infection in Human Cells by Alpha, Beta, and Gamma Interferons

doi:

Figure Lengend Snippet: Effect of IFN pretreatment on antibody-dependent DV2 infection of THP-1 and U937 cells. THP-1 and U937 cells were exposed to DV2 (strain 16681) at an MOI of 10 in the absence (No Ab) or presence (4G2) of an enhancing MAb. After incubation for 96 h, cells were processed for flow cytometry (A), and supernatants were harvested for plaque assays (B). In each case, cells were pretreated 24 h prior to exposure to virus with medium, IFN-α (100 IU/ml), IFN-β (10 ng/ml), IFN-γ (10 ng/ml), or IFN-β+γ (each at 10 ng/ml). The data are expressed as in Fig. ​Fig.2.2. One representative experiment of three is shown.

Article Snippet: Murine hybridomas against human DV antigens (3H5-1, anti-DV2; 5D4-11, anti-DV3; and 2H2-9, anti-DV) or flavivirus antigens (4G2) were obtained from the ATCC and grown in Dulbecco's modified Eagle medium (Gibco BRL) supplemented with 10% FBS, 2 mM l -glutamine, 1 mM sodium pyruvate, penicillin G (100 U/ml), and streptomycin (100 μg/ml) at 37°C in 5% CO 2 .

Techniques: Infection, Incubation, Flow Cytometry

Journal:

Article Title: Modulation of Dengue Virus Infection in Human Cells by Alpha, Beta, and Gamma Interferons

doi:

Figure Lengend Snippet: Time course of the effect of IFN pretreatment on antibody-dependent DV2 infection of K562 cells. K562 cells were exposed to DV2 (strain 16681) at an MOI of 0.005 in the presence of an enhancing MAb (3H5). After incubation for 96 h, cells (106) were processed for flow cytometry. In each case, cells were exposed to medium, IFN-α (100 IU/ml), IFN-β (10 ng/ml), IFN-γ (10 ng/ml), or a combination of IFNs as described for Fig. ​Fig.2.2. One representative experiment of two is shown.

Article Snippet: Murine hybridomas against human DV antigens (3H5-1, anti-DV2; 5D4-11, anti-DV3; and 2H2-9, anti-DV) or flavivirus antigens (4G2) were obtained from the ATCC and grown in Dulbecco's modified Eagle medium (Gibco BRL) supplemented with 10% FBS, 2 mM l -glutamine, 1 mM sodium pyruvate, penicillin G (100 U/ml), and streptomycin (100 μg/ml) at 37°C in 5% CO 2 .

Techniques: Infection, Incubation, Flow Cytometry

Journal:

Article Title: Modulation of Dengue Virus Infection in Human Cells by Alpha, Beta, and Gamma Interferons

doi:

Figure Lengend Snippet: IFN effect on the levels of positive and negative strands of DV2 RNA in HepG2 cells. HepG2 cells were exposed to DV2 (strain 16681) at an MOI of 2 and incubated for 24 h. In each case, cells were exposed to medium, IFN-γ (10 ng/ml), IFN-β (10 ng/ml), or IFN-β+γ (each at 10 ng/ml) 24 h prior to incubation with virus. Cells (6 × 105) were harvested, total RNA was isolated, and quantitative asymmetric RT-PCR was performed with fixed amounts of cellular RNA in the presence of 10-fold decreasing concentrations of positive- or negative-strand competitor. The RT-PCR product was subjected to agarose gel electrophoresis. M refers to the molecular weight marker, C denotes an RT-PCR with only competitor RNA, and the number above each lane represents the log number of copies of competitor used. The amount of viral RNA was determined from the competitor concentration that produces competitor and DV bands of equal intensity (denoted by asterisk). RNA per cell is calculated as defined in Materials and Methods. For the positive strand, the equivalence point and RNA copies per cell were as follows: media, 107 and 833; IFN-γ, 5 × 106 and 417; IFN-β, 104 and 0.8; IFN-β+γ, 104 and 0.8. For the negative strand, the corresponding values were as follows: media, 106 and 110; IFN-γ, 105 and 11; IFN-β, 103 and 0.1; IFN-β+γ, undetectable.

Article Snippet: Murine hybridomas against human DV antigens (3H5-1, anti-DV2; 5D4-11, anti-DV3; and 2H2-9, anti-DV) or flavivirus antigens (4G2) were obtained from the ATCC and grown in Dulbecco's modified Eagle medium (Gibco BRL) supplemented with 10% FBS, 2 mM l -glutamine, 1 mM sodium pyruvate, penicillin G (100 U/ml), and streptomycin (100 μg/ml) at 37°C in 5% CO 2 .

Techniques: Incubation, Isolation, Reverse Transcription Polymerase Chain Reaction, Agarose Gel Electrophoresis, Molecular Weight, Marker, Concentration Assay

Journal:

Article Title: Modulation of Dengue Virus Infection in Human Cells by Alpha, Beta, and Gamma Interferons

doi:

Figure Lengend Snippet: Time course of IFN effect on the levels of positive and negative strands of DV2 RNA in HepG2 cells. HepG2 cells were exposed to DV2 (Thai strain K0049) at an MOI of 2 and incubated for 72 h. In each case, cells were exposed to medium or IFN-β+γ (each at 10 ng/ml) either 24 or 4 h before (pre) or after (post) incubation with virus. Cells (6 × 105) were harvested, total RNA was isolated, and asymmetric RT-PCR was performed with fixed amounts of cellular RNA in the presence of 10-fold decreasing concentrations of positive- or negative-strand competitor. The amount of viral RNA per cell (A) was determined after agarose gel electrophoresis (B) as described for Fig. ​Fig.66.

Article Snippet: Murine hybridomas against human DV antigens (3H5-1, anti-DV2; 5D4-11, anti-DV3; and 2H2-9, anti-DV) or flavivirus antigens (4G2) were obtained from the ATCC and grown in Dulbecco's modified Eagle medium (Gibco BRL) supplemented with 10% FBS, 2 mM l -glutamine, 1 mM sodium pyruvate, penicillin G (100 U/ml), and streptomycin (100 μg/ml) at 37°C in 5% CO 2 .

Techniques: Incubation, Isolation, Reverse Transcription Polymerase Chain Reaction, Agarose Gel Electrophoresis

Journal:

Article Title: Modulation of Dengue Virus Infection in Human Cells by Alpha, Beta, and Gamma Interferons

doi:

Figure Lengend Snippet: Time course of the levels of positive and negative strands of DV2 RNA after IFN treatment of HepG2 cells. HepG2 cells were exposed to DV2 (strain 16681) at an MOI of 2 and incubated as indicated prior to harvest. In each case, cells were pretreated with medium or IFN-β+γ (each at 10 ng/ml) for 24 h prior to exposure to virus. At each time point after infection, cells were harvested and counted by hemocytometry, and total RNA was isolated. Subsequently, asymmetric RT-PCR was performed with fixed amounts of cellular RNA in the presence of 10-fold decreasing concentrations of positive- or negative-strand competitor. The product was subjected to agarose gel electrophoresis, and the amount of viral RNA per cell was determined as described for Fig. ​Fig.66.

Article Snippet: Murine hybridomas against human DV antigens (3H5-1, anti-DV2; 5D4-11, anti-DV3; and 2H2-9, anti-DV) or flavivirus antigens (4G2) were obtained from the ATCC and grown in Dulbecco's modified Eagle medium (Gibco BRL) supplemented with 10% FBS, 2 mM l -glutamine, 1 mM sodium pyruvate, penicillin G (100 U/ml), and streptomycin (100 μg/ml) at 37°C in 5% CO 2 .

Techniques: Incubation, Infection, Isolation, Reverse Transcription Polymerase Chain Reaction, Agarose Gel Electrophoresis