Clonal expansion is severely compromised in MyD88−/− and TLR2−/− clone 4 CD8 T cells in response to viral infection in vivo. A total of 104 naive WT, MyD88−/−, or TLR2−/− clone 4 CD8 T cells (Thy1.1+) were transferred into WT (WT→WT, MyD88−/− →WT, or TLR2−/−→WT) or MyD88−/− (WT→MyD88−/−) recipient mice (Thy1.2+) that were either left uninfected (naive) or infected with 5 × 105 pfu rVV-HA intraperitoneally. Seven days after infection, splenocytes were analyzed for the expansion of clonotypic T cells and their function by IFN-γ intracellular staining. (A) The percentages of clonotypic T cells among total lymphocytes (top) and IFN-γ–producing clonotypic T cells among total CD8+ T cells (bottom) in rVV-HA infected mice are indicated. (B) The absolute cell number per spleen of CD8+Thy1.1+ clonotypic T cells in both uninfected and rVV-HA infected hosts with SDs are indicated (n = 4 per group). For all groups: naive versus rVV-HA, P < .001. For rVV-infected hosts: WT→MyD88−/− versus WT→WT, P < .05; MyD88−/−→WT or TLR2−/−→WT versus WT→WT, P < .001. (C) The mean fluorescence intensity (MFI) of IFN-γ–producing clonotypic cells is indicated. WT→MyD88−/−, MyD88−/−→WT, or TLR2−/−→WT versus WT→WT, P > .05. Data shown are representative of 3 independent experiments.