Discrimination between brain parenchyma-localized and vasculature-localized lymphocyte proliferation.

Foxp3-DTR, MCMV-infected, DTx-treated and untreated mice at 14 dpi were injected intravenous with anti-CD8α-PE and anti-CD4-FITC mAb. Lymphocytes were isolated and stained ex-vivo for the anti-CD8 β-AF647 and anti-CD4-AF700 using different clones, as described in the methods. Plots are representative of two experiments using three animals per groups. (A) Contour plots show CD8+ T-cells in the vasculature that stained both for anti-CD8α-PE and anti-CD8β-AF647; while tissue lymphocytes were stained by anti-CD8β-AF647 alone in both DTx-treated and untreated groups. (B) Contour plots show proliferation of CD8+ T-cells both within the tissue and in the vasculature. C. Contour plots represent proliferation of CD4+ T-cells both in tissue and vasculature. (D) The number of parenchyma-localized CD8+ T-cells within MCMV-infected brains of animals with and without DTx treatment is shown. (E) The number of parenchymal CD4+ T-cells with and without DTx treatment is shown. **p < 0.001 DTx- versus DTx+ animals