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

Article Title: Human cytomegalovirus differentially controls B cell and T cell responses through effects on plasmacytoid dendritic cells.

doi: 10.4049/jimmunol.179.11.7767

Figure Lengend Snippet: FIGURE 2. HCMV activates PDCs. A, HCMV alters PDC morphology. PDCs were incubated overnight with medium or TB40/E at an MOI of 5 and examined by light microscopy and Evans blue staining of cytospin preparations. At 24 h, mock-infected PDCs showed plasmacytoid morphology, whereas TB40E-infected PDCs formed clusters, developed dendrites, and had larger cytoplasm. B and C, HCMV induces partial maturation of PDCs. PDCs were mock-infected (mock inf), stimulated with CpG-A (3 g/ml), or infected with TB40/E or VR1814 (MOI of 5) overnight. The stimuli were removed, cells were washed, and fresh medium was added. After 24 h, expression of CD83, HLA-DR, BDCA-2, CD80, and CD86 was measured by FACS. B, Values are means SEM of seven donors in independent experiments. , p 0.05. Y-axis values indicate MFI. C, Open histograms depict cells stained with isotype-matched control Ab. Representative data from one of seven experiments are shown.

Article Snippet: For cell surface staining, we used Abs against CD1a, CD3, CD4, CD8, CD14, CD19, CD20, CD38, CD80, CD86, HLA-DR, and mouse isotype controls (BD Pharmingen); CD83 (Immunotech); and BDCA-2 and BDCA-4 (Miltenyi Biotec).

Techniques: Incubation, Light Microscopy, Staining, Infection, Expressing, Control

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

Article Title: Human cytomegalovirus differentially controls B cell and T cell responses through effects on plasmacytoid dendritic cells.

doi: 10.4049/jimmunol.179.11.7767

Figure Lengend Snippet: FIGURE 4. Soluble factors secreted by HCMV-infected DCs contribute to B cell activation. A and B, PDCs were mock infected or infected with TB40/E (MOI of 5) for 4 h, washed, and resuspended in complete medium for 18 h. Supernatants were collected and filtered through 0.1-m filters to eliminate viral particles. B cells (2.5 105 cells/well) were cultured in 200 l of supernatant from PDC cultures in 96-well round-bottom plates. B cells were unstimulated or stimulated by B cell-receptor ligation (anti-Ig, 10 g/ml). A, After 3 days, B cells were harvested, stained for the B cell markers CD19, CD20, CD38, and CD86, and analyzed by FACS. Values are means SD. , p 0.05 (n 4 donors in independent experiments). B, For proliferation assay, B cells were labeled with 1 M CFSE, incubated with PDC supernatant for 5 days, harvested, and analyzed by FACS. Data from one representative of three donors in independent experiments are shown. C, MDCs were mock infected or infected with TB40/E (MOI of 5) for 4 h, washed, and resuspended in complete medium for 18 h. Supernatants were collected and filtered. B cells (2.5 105 cells/well) were cultured in 200 l of supernatant from MDC cultures. B cells were unstimulated or stimulated by B cell-receptor ligation (anti-Ig, 10 g/ml). After 3 days, B cells were harvested, stained for the B cell markers CD19, CD20, CD38, and CD86, and analyzed by FACS. Values are means SD. , p 0.05 (n 4 donors in independent experiments).

Article Snippet: For cell surface staining, we used Abs against CD1a, CD3, CD4, CD8, CD14, CD19, CD20, CD38, CD80, CD86, HLA-DR, and mouse isotype controls (BD Pharmingen); CD83 (Immunotech); and BDCA-2 and BDCA-4 (Miltenyi Biotec).

Techniques: Infection, Activation Assay, Cell Culture, Ligation, Staining, Proliferation Assay, Labeling, Incubation

Journal: Retrovirology

Article Title: The efficiency of Vpx-mediated SAMHD1 antagonism does not correlate with the potency of viral control in HIV-2-infected individuals

doi: 10.1186/1742-4690-10-27

Figure Lengend Snippet: Effect of HIV-1 and HIV-2 on DC activation. ( A ) CD86 surface expression in DCs after infection with HIV-1 and HIV-2 IRES-eGFP constructs alone or in combination with VSV-G-pseudotyped SIVmac239 particles. The levels of CD86 expression by virally infected (eGFP+) cells were measured at 3 days post-infection. Panels A and B show mean values (±SEM) derived from four experiments. HIV-1 alone did not infect DCs at detectable levels. ( B ) Levels of IFN-γ in the supernatant of the infected DC cultures.

Article Snippet: DC activation was measured using the APC conjugated monoclonal antibody against CD86 (5397868, Invitrogen) by flow cytometry.

Techniques: Activation Assay, Expressing, Infection, Construct, Derivative Assay