Figure 1.

HIV Capture Kinetics and Localization by Mature DCs in 3D Collagen

(A) Schematic illustration of the HIV-iGFP/dTomato reporter vector. PR, protease cleavage site.

(B) Immature or LPS-stimulated MDDCs were incubated with HIV-iGFP/dTomato for the indicated times and viral capture kinetics assessed by flow cytometry. Numbers indicate % of DCs that captured HIV-iGFP particles.

(C) Time course analysis of HIV-iGFP capture by DC populations. Representative data from two (immature DC) and three (mature DC) independent experiments are shown. Mean ± SEM.

(D) A time series micrograph of immature or LPS-stimulated DCs exposed to HIV-iGFP in collagen for the indicated times. Time stamp in min:sec represents elapsed time of the recordings and not the start of HIV incubation. Right panels: 3D surface rendering of DCs and HIV particles, with XY, YZ, and XZ views that illustrate viral particle distribution.

(E) Representative line profile analysis of GFP fluorescence intensity in HIV-captured DC, from the leading to the trailing edge of each cell. Representative “HIV capture” and “HIV compartmentalization” line profiles are shown. HIV compartmentalization was defined as >70% of total GFP fluorescence that is confined within the training edge of polarized DCs. Relative proportion of HIV found compartmentalized over time is shown. Data from two independent experiments are shown (n = 303 total cells).