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. 2013 Feb;26(1):109–113. doi: 10.1089/vim.2012.0052

FIG. 1.

FIG. 1.

VP2.139 peptide-immunized NOD, SJL, and B6 mice generate Q9:VP2.139-specific CD8 T cell responses. (A) Representative tetramer staining of splenocytes from B6.Kb-/-Db-/- mice 3 wk after MPyV inoculation. RBC-lysed splenocytes were incubated with FcBlock, then surface stained with CD8α (clone 53–6.7), CD44, and either Q9:VP2.139 tetramer or a chimeric Q9(Kb):VP2.139 tetramer. Splenocytes gated on CD8+ CD44+ cells. Geometric MFI of the tetramer-associated fluorophore within the gated population is indicated. Data are representative of 10 mice pooled from two separate experiments. (B) Representative Q9:VP2.139 tetramer staining of splenocytes from VP2.139 peptide-immunized NOD mice at day 6 p.i. by MPyV. “VP2.139” indicates mice immunized with CFA+peptide. “No Peptide” indicates control mice receiving CFA+PBS. Q9:L19 is the control tetramer for Q9:VP2.139. Numbers indicate the frequency of tetramer+ CD8+ lymphocytes. Data are representative of 11 mice pooled from three separate experiments. (C) Frequency of Q9:VP2.139-specific CD8+ lymphocytes, with Q9:L19 tetramer staining subtracted, in VP2.139 peptide-immunized NOD (circles), SJL (triangles), and B6 (diamonds) mice at day 6 p.i. Horizontal lines indicate geometric mean. Symbols represent individual mice. Filled symbols indicate VP2.139-primed mice. Open circles indicate PBS-primed mice. Two NOD mice with no detectable Q9:VP2.139 tetramer staining were omitted from the phenotyping in Figure 1D. (D) Phenotyping of splenic Q9:VP2.139-specific CD8 T cells in NOD and SJL mice. Q9:VP2.139 tetramer+ CD8 T cells were surface-stained with antibodies to the indicated molecules. Data are clustered by costained markers, as indicated in the y axis. “+” indicates cells which are positive for the indicated molecule, and “−” indicates cells which are negative for the indicated molecule. “hi” indicates cells expressing a high level of the indicated molecule, and “lo” indicates cells expressing a low level of the indicated molecule. Data are representative of 9 mice (NOD) and 10 mice (SJL) pooled from two separate experiments.