Journal: Virology Journal

Article Title: Herpes simplex virus type 2 infection increases human immunodeficiency virus type 1 entry into human primary macrophages

doi: 10.1186/1743-422X-8-166

Figure Lengend Snippet: HSV-2 ability to infect human monocyte cell line U937 relies on their differentiation level . A) HSV-2 replication in undifferentiated U937 cells. The U937 cell line was infected with two different MOI of HSV-2, as reported in the graph legend. Vero cells were infected as a control. HSV-2 titre in the cellular supernatants was measured by plaque assay, at different times post-infection (p.i.). B) Effect of TPA treatment of U937 cells on CD14 expression. The effect of TPA treatment on U937 cell differentiation state was analyzed by CD14 FACS analysis. The percentage of positive cells is reported. C) HSV-2 replication in TPA-treated U937 cells. Undifferentiated or TPA-treated U937 cells were infected with HSV-2 (MOI of 1 PFU/cell). HSV-2 titre in the cellular supernatant was measured by plaque assay. In all cases, the reported values represent the mean of four independent experiments. The error bars represent the standard deviation.

Article Snippet: Briefly, 1 × 10 6 cells were harvested and directly incubated for one hour in cold PBS containing 1:100 (v/v) of an anti-human CD14 primary antibody (Li StarFISH).

Techniques: Infection, Control, Plaque Assay, Expressing, Cell Differentiation, Standard Deviation

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: An Allosteric shift in CD11c affinity activates a pro-atherogenic state in arrested intermediate monocytes

doi: 10.4049/jimmunol.2000485

Figure Lengend Snippet: Integrin expression is elevated on monocytes from blood samples of cardiac patients compared with healthy age-matched subjects. (A) Gating strategy for the monocyte subsets based on expression of HLA-DR, CCR2, CD14, and CD16 applied to each patient cohort to define monocyte subsets, as indicated via representative images (CD14, green; CD16, red). Blood was collected from age-matched healthy controls (n = 6) and compared with CAD (n = 16) and NSTEMI (n = 27) on the day of percutaneous intervention for assessment of integrin expression on (B) cMo, (C) iMo, and (D) ncMo via FACS. Data are represented as mean receptor number ± SD (two-way ANOVA with Tukey posttest, *p < 0.05 between healthy age-matched subject cohorts).

Article Snippet: Monocyte subset abundance was quantified by calibrating fluorescent signals derived from the CD14, CD16, and CCR2 primary Ab conjugates to receptor expression calculated using Bangs Laboratories beads with a known number of binding sites/area.

Techniques: Expressing

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: An Allosteric shift in CD11c affinity activates a pro-atherogenic state in arrested intermediate monocytes

doi: 10.4049/jimmunol.2000485

Figure Lengend Snippet: Monocyte recruitment to inflamed endothelium under shear stress is β1- and β2-integrin–dependent on binding to VCAM-1. (A) Monocytes were perfused into the A-Chip onto TNF-α–stimulated HAEC in the presence of blocking Abs to CD11c, CD11b, VLA-4, or a nonspecific isotype IgG (Control) and TEM of adherent cells (white arrows) assessed by phase-contrast imaging after 10 min of shear at 2 dynes/cm2. Images display a time course of monocyte arrest and transmigration (phase dark) over 2 min. Scale bar, 10 μm. Monocyte subsets were determined based upon on-chip analysis of relative fluorescence intensity of Abs to CD14, CD16, and CCR2, which enabled the assessment of the number of arrested per field of view (FOV) at original magnification ×40. (B) Quantification of arrested iMo (filled bars) and the number of those arrested cells that have transmigrated (open bars) over 10 min of constant shear (2 dynes/cm2). Data are represented as mean arrested or transmigrated cells per FOV ± SD (n = 4–5 per group) (two-way ANOVA with Tukey posttest *p < 0.05 between isotype control within each cohort TEM color, #p < 0.05 compared with healthy, $p < 0.05 between CAD and NSTEMI cohorts for each Ab blocking treatment).(C) Monocyte subset arrest frequency on rVCAM-1 on the A-Chip. Data are represented as mean ± SD monocyte subset fraction of total arrested measured on the day of percutaneous coronary intervention or venous blood draw from healthy (n = 6), CAD (n = 8), and NSTEMI (n = 20) analyzed via two-way ANOVA with Tukey posttest. *p < 0.05 between healthy age-matched subject groups.

Article Snippet: Monocyte subset abundance was quantified by calibrating fluorescent signals derived from the CD14, CD16, and CCR2 primary Ab conjugates to receptor expression calculated using Bangs Laboratories beads with a known number of binding sites/area.

Techniques: Shear, Binding Assay, Blocking Assay, Control, Imaging, Transmigration Assay, Fluorescence