Fig. 6
Neurons are the sites of latent and reactivating MCMV in the CNS. A-C, F Newborn R26tdTomato mice were infected with MCMV-GFP-Cre i.p. A Brains were collected and analyzed 45 dpi. A representative brain section (10x magnification) containing tdTomato-positive cells (left) and representative magnified (63x magnification) tdTomato-positive cell (right) are shown. B Representative image of tdTomato-positive NeuN+ neuron (40x magnification). C Quantification of cell types expressing tdTomato reporter protein 45 dpi (n = 5 mice, 5 sections/mouse). ND - Not detected. D Newborn C57BL/6 mice were infected with MCMV. Brain regions were isolated and cultured ex vivo for 6 weeks. The reactivation frequency in indicated brain regions was determined (n = 6 mice). ND - not detected. E Newborn C57BL/6 mice were infected with MCMV. The number of CD4+ T-cells per gram of tissue was analyzed using flow cytometry at 180 dpi. Columns indicate median values (n = 6–10 mice). Kruskal–Wallis test was used was used. F Mice were depleted of CD4+ T-cells starting on 90 dpi for 1 month. Mice were perfused, sacrificed and brains were collected and frozen. Quantification of co-expression of NeuN and tdTomato is shown (n = 3–5 mice, 5 sections/mouse). Columns indicate median values. Mann–Whitney two-tailed test was used. G Newborn C57BL/6 mice were infected with MCMV and mice were depleted of CD4+ T-cells starting at 90 dpi for 1 month (n = 10 mice). Brain regions were separated, homogenized and layered on MEF cells in multiwall plates. The frequency of MCMV-positive wells was determined. Results for individual mice are shown (circles, squares and rectangles). The number of mice from which the virus was recovered is indicated above the graph. H-I Newborn C57BL/6 mice were infected with MCMV and mice were depleted of CD4+ T-cells starting on 90 dpi for 1 month. H Representative image of MCMV-infected MAP2+ neuron. The arrow points to the MCMV-infected neuron (40 magnification, 100x magnification insert). I Quantification of IE1 protein-positive MAP2 neurons (n = 4 mice, 20 sections/mouse). p values statistically indicate significant differences