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Figure S1. Purity and gating strategy for FACs sorting (A) CD4+ T cells were isolated from spleens of naive CD45.1+ M25-II mice by positive MACS separation. A representative example of the purity of MACS-sorted cells is shown; cells are gated on live lymphocytes. 105 MACS-sorted cells were transferred into CD45.2+ C57BL/6 recipients followed by MCMV infection. 8 days post infection, CD45.1+ M25-II TCR tg CD4+ T cells were FACS sorted from the lung. (B) Gating strategy for FACS sorting. Lung lymphocytes were stained for CD4 and CD8 and within CD4+ T cells CD45.1+ were sorted. A representative stainings are shown. (C) Purity of FACS sorted cells. A representative sample is shown.

Figure S2. Activated LCMV-specific CD4+ T cells do not protect immune-compromised mice from cytopathogenic CMV infection CD4+ T cells were isolated from spleens of naive CD45.1+ SMARTA mice by positive MACS separation and 104 LCMV-specific CD4+ T cells were transferred to B6 (CD45.2+) recipient mice and were infected intravenously with 200 PFU LCMV-WE one day after the cell transfer. LCMV-specific CD4+ T cells (CD45.1+) were isolated from spleen leukocytes at day 8 post infection by FACS sorting and adoptively transfused into sublethally irradiated B6 recipient mice. The recipient mice were subcutaneously infected 1 day post transfer with 105 PFU Δm157 MCMV. The virus titers were determined in spleen, liver, lung and salivary gland at day 12 post infection. Data are shown from one experiment. Each symbol represents one individual mouse, horizontal line indicates the mean, dashed line indicates detection limit. Statistical analysis was performed by 2-tailed unpaired student's t-test (* p < 0.05, ** p < 0.01, *** p < 0.001).

Figure S3. M38-specific CD8+ T cells provide protection from pathogenic CMV infection in various peripheral tissues but not in the salivary gland CD8+ T cells were isolated from spleens of naive CD45.1+ MAXI mice by positive MACS separation and 104 M38-specific CD8+ T cells were transferred to B6 (CD45.2+) recipient mice and infected intravenously with 5×106 PFU Δm157 MCMV one day after the cell transfer. At day 8 post infection, M38-specifc CD8+ T cells (CD45.1+) were isolated from lung leukocytes by FACS sorting and adoptively transfused in graded numbers into sublethally irradiated recipient B6 mice. The recipient mice were subcutaneously infected 1 day post transfer with 105 PFU Δm157 MCMV. The virus titers were determined in spleen, liver, lung and salivary gland at day 12 post infection. Each symbol represents one individual mouse, horizontal line indicates the mean, dashed line indicates detection limit. Data are shown from one experiment. Statistical analysis was performed by 2-tailed unpaired student's t-test (* p < 0.05, ** p < 0.01, *** p < 0.001).

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