Journal: The American Journal of Tropical Medicine and Hygiene

Article Title: Dengue Virus Infection of Primary Human Smooth Muscle Cells

doi: 10.4269/ajtmh.18-0175

Figure Lengend Snippet: Human umbilical artery smooth muscle cells (HUASMC) are permissive to dengue virus (DENV) infection. Virion production and release was quantified by plaque assays of culture supernatants at different time points post-infection (p.i.) of HUASMC, human umbilical vein endothelial cells (HUVEC), and LLC-MK2 (macaque kidney cells) infected with each of the four DENV serotypes (multiplicity of infection:1). (A) Infection kinetics of HUASMC cells (24–72 hours p.i.) with DENV 1–4 measured by plaque assays (plaque-forming units [PFU]/mL). (B) DENV 1–4 titers in culture supernatants of HUASMC, HUVEC, and LLC-MK2 cells at 72 hours p.i. Data are expressed as the mean ± standard deviation of three independent experiments. ***P < 0.001 compared with its HUVEC and LLC-MK2 cells counterparts. (D) l. = assay detection limit.

Article Snippet: Human umbilical artery smooth muscle cells and HUVEC were purchased and maintained in smooth muscle cell growth medium and endothelial cell growth medium, respectively, according to the manufacturer’s instructions (Cell Applications, San Diego, CA).

Techniques: Virus, Infection, Standard Deviation

Journal: The American Journal of Tropical Medicine and Hygiene

Article Title: Dengue Virus Infection of Primary Human Smooth Muscle Cells

doi: 10.4269/ajtmh.18-0175

Figure Lengend Snippet: Human umbilical artery smooth muscle cells (HUASMC) release dengue virus (DENV) genomes on infection. Dengue virus genomic RNA was quantified by real-time qRT-PCR from cell culture supernatants of HUASMC, human umbilical vein endothelial cells (HUVEC), and LLC-MK2 (macaque kidney cells) infected with DENV (multiplicity of infection:1). (A) Dengue virus 1–4 infection kinetics (24–72 hours post-infection [p.i.]) of HUASMC cells measured by real-time genomic qRT-PCR (copies/mL). (B) Genome copies present in culture supernatants of HUASMC, HUVEC, and LLC-MK2 cells at 72 hours p.i. with the four DENV serotypes. (C) Calculated genome-to-plaque-forming unit (PFU) ratios of HUASMC, HUVEC, and LLC-MK2 cells supernatants at 72 hours p.i. with each DENV serotype. Data are expressed as the mean ± standard deviation of three independent experiments. *P < 0.05, **P < 0.005, and ***P < 0.001 calculated to its HUASMC counterpart.

Article Snippet: Human umbilical artery smooth muscle cells and HUVEC were purchased and maintained in smooth muscle cell growth medium and endothelial cell growth medium, respectively, according to the manufacturer’s instructions (Cell Applications, San Diego, CA).

Techniques: Virus, Infection, Quantitative RT-PCR, Cell Culture, Standard Deviation

Journal: The American Journal of Tropical Medicine and Hygiene

Article Title: Dengue Virus Infection of Primary Human Smooth Muscle Cells

doi: 10.4269/ajtmh.18-0175

Figure Lengend Snippet: Dengue virus (DENV) antigens are detected in human umbilical artery smooth muscle cells (HUASMC). Epifluorescence images of immunostained cells with an anti-DENV 1-4 envelope protein-specific monoclonal antibody (green), and a cytoplasmic (red) and nuclei (blue) counterstains. The images were captured at 72 hours post-infection (p.i.) with each DENV serotype at a multiplicity of infection (MOI) of 1. (A) Representative image of mock-infected HUASMC cells at ×100 magnification (scale bar = 50 μm). (B and C) Representative images of DENV-infected HUASMC (arrows) at ×400 (scale bar = 20 μm) and ×100 (scale bar = 50 μm) magnification, respectively. (D) Image analysis for quantifying infected cells (green outline) vs total cell nuclei (white outlines) with the software CellProfiler 2.0. (E) Percentages of infected cells in HUASMC, human umbilical vein endothelial cells (HUVEC), and LLC-MK2 (macaque kidney cells) cell lines. (F) Comparison of DENV labeling by immunostaining with an anti-DENV 1-4 envelope protein-specific monoclonal antibody (green) and an anti-NS3 polyclonal antibody (red) in HUASMC cells at 72 hours p.i. with DENV-2 and DENV-3 at a MOI of 1. Magnification of ×400 (scale bar = 20 μm). Data are expressed as mean ± standard deviation of three independent experiments. **P < 0.005, ***P < 0.001 calculated to its HUASMC cells counterpart. This figure appears in color at www.ajtmh.org.

Article Snippet: Human umbilical artery smooth muscle cells and HUVEC were purchased and maintained in smooth muscle cell growth medium and endothelial cell growth medium, respectively, according to the manufacturer’s instructions (Cell Applications, San Diego, CA).

Techniques: Virus, Infection, Software, Comparison, Labeling, Immunostaining, Standard Deviation