A Prominent Role for DC-SIGN+ Dendritic Cells in Initiation and Dissemination of Measles Virus Infection in Non-Human Primates
Figure 5
DC-SIGN expressed by BAL and lymph node cells enhances MV transmission.
(A) DC-SIGNhi/HLA-DR+ (p1), DC-SIGNlo/HLA-DR+ (p2) and HLA-DR−/DC-SIGN- (p3) cells were sorted by FACS from BAL and lymphoid tissues of uninfected animals. Gates and percentages of the subsets are depicted. A representative FACS plot of 3 independent experiments is shown. (B) DC-SIGN on sorted BAL cells binds HIV-1 gp120-coated fluorescent beads. Binding was blocked by mannan (0.25 mg/ml). Representative data of 2 independent experiments are shown. (C) CD150 expression of the 3 subsets from BAL and TBLN (black lines) compared to isotype controls (gray areas). Percentages of CD150+ cells are depicted in the histograms. (D) Sorted cells from lymph nodes of an uninfected animal were incubated with fluorescently labeled rMVKSEGFP or medium control. Binding was measured by flow cytometry and the mean fluorescent intensity (MFI) is depicted. (E) Representative example of 2 indepedent ex vivo infections of BAL cells with rMVKSEGFP (MOI 3) 24 hours post infection. The left (brightfield) and right (EGFP fluorescent) panel are corresponding pictures. (F) Infection of sorted DC-SIGN subsets with rMVKSEGFP (MOI 3) was determined by measuring EGFP in FACS. Means and standard deviations of 2 independent experiments are shown. (G-H) Cells were incubated with rMVKSEGFP (MOI 1) for 3 hours. Then the cells were washed and B cells were added. After 24 hours EGFP expression was measured by flow cytometry. For BAL cells, combined data of 2 independent experiments are shown (G). Cells isolated from lymph nodes were pre-incubated with blocking anti-DC-SIGN antibodies (20 ug/ml) for 30 minutes. ** p<0.01 (H). Bars represent the mean of duplicates.