Journal: Journal of Virology
Article Title: Sensing of SARS-CoV-2-infected cells by plasmacytoid dendritic cells is modulated via an interplay between CD54/ICAM-1 and CD11a/LFA-1 α L integrin
doi: 10.1128/jvi.01235-24
Figure Lengend Snippet: Sensing of SARS-CoV-2-infected cells by pDCs is more efficient compared with cell-free virions. ( A ) Quantification of IFN-I (IFN-α/β) in the supernatant of PBMCs that were co-cultured for 24 h with SARS-CoV-2 (LSPQ1)-infected HeLa-hACE2, exposed to cell-free SARS-CoV-2 at different MOIs, or treated with supernatant of infected cells (MOI 0.1) (sup) collected at 24 h post-infection. PBMCs used in the co-cultures and in “cell-free virus exposure” conditions were from the same donors with the same number of cells. In the latter conditions, the MOI was based on the number of PBMCs. UI, uninfected cells. Each dot represents independent experiments/donors. Mann-Whitney U test. ** P < 0.01. ( B ) (Upper panel) Frequency of SARS-CoV-2 infection in HeLa-hACE2 as determined by flow cytometry using αN Ab and a mouse IgG1 isotype control (ctrl). (Lower panel) A summary dot plot shows the results from six independent infections. ( C ) Quantification of IFN-I in the supernatant of SARS-CoV-2-infected HeLa-hACE2 (MOI 0.1, 24 h) that were co-cultured with pDC-depleted PBMCs or total PBMCs ( n = 3 donors). ( D ) The analysis was as described for panel C but with cell-free viruses instead of infected cells ( n = 2 donors). The MOI (0.1) used was based on the number of PBMCs, as done in . In all relevant panels, the co-cultures were done for 24 h. Shown is the mean ± SEM. The dotted line indicates the limit of detection (LOD) of the assay (60 U/mL). See also .
Article Snippet: Cells were then stained with the mouse αN primary Ab (1/200, Sino Biologicals) or the isotype control mouse IgG1 (1/200, BioLegend) for 30 min at room temperature, followed by staining with a secondary Ab for 30 min at room temperature.
Techniques: Infection, Cell Culture, Virus, MANN-WHITNEY, Flow Cytometry, Control