RIP-OVA mice were injected with Rag-2-deficient OT-I cells, and immunization with peptide and LPS followed.
(A) Surface expression of CD25, CD69, LFA-1 (CD11a), and CD8 on lymph node OT-I T cells after 8 hr (n = 4 per peptide).
(B) CFSE dilution profiles of lymph node OT-I T cells 3 days after peptide immunization (n = 3 per peptide). The numbers of recovered OT-I donor cells from spleen and lymph nodes were equal to 2.2 ± 0.3 × 106 and 0.7 ± 0.1 × 106 for Q4R7 and Q4H7 immunized mice, respectively.
(C) Total numbers of splenic OT-I T cells in mice adoptively transferred with 3 × 104 OT-Is and 1 day later infected with Lm-Q4R7 or Lm-Q4H7 (two experiments, five mice per peptide).
(D) IFN-γ production by OT-I T cells restimulated for 5 hr with OVA peptide in vitro (n = 6 per peptide).
(E) CTL activity from OT-I T cells. OVA (CFSEhi) and VSV (CFSElo) peptide-pulsed splenocytes were mixed at a 1:1 ratio and injected into RIP-OVA mice 3 days after peptide immunization. Target cell lysis was analyzed from splenocytes 5 hr after cell transfer. CTL activity from unimmunized mice was <10% (n = 7, Q4R7; n = 6 Q4H7). Representative data are shown for n ≥ 2 separate experiments. Error bars denote standard error of the mean (SEM).