Figure 5

Detection of GFP⁺ cells in ferrets after IM injection and IT inoculation of rMVA-GFP. (a) Direct detection of GFP⁺ cells using CLSM after IM injection (left panel) or IT inoculation (middle and right panel) of 10⁹ PFU rMVA-GFP, 6 HPA. Middle panel shows direct CLSM of a lung slice, whereas the right panel shows cells recovered from the lungs by BAL. Contrast of certain images was linearly enhanced using Adobe Photoshop CC. (b) Overview of detection of GFP⁺ cells in BAL (left [L] or right [R] lung lobes), TB-LN, ING-LN, WBC or spleen in each individual ferret (n = 2 per group) after IM injection or IT inoculation of rMVA-GFP. Green = GFP⁺. Red = GFP-negative. (c) Frequency of GFP⁺ cells in the respective tissues after IM injection or IT inoculation of rMVA-GFP. Number of GFP⁺ cells per 10⁶ total cells is shown, mean of two ferrets is indicated. (d) GFP⁺ cells were phenotypically characterized as DC-like, monocyte-like or lymphocyte-like cells based on scatter properties of the different cell types.