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. 2011 Aug 1;208(8):1605–1620. doi: 10.1084/jem.20102101

Figure 2.

Figure 2.

CXCR3−/− memory CD8+ T cells exhibit similar functional capacities to WT cells. (A–C) Mixed CXCR3−/− and WT OT-I cells were transferred, followed by VV-OVA infection as in Fig.1 B. Cytokine production and granzyme B expression by CXCR3−/− and WT memory OT-I cells was analyzed at day 43 after infection. (A) Representative flow cytometry plots showing TNF, IL-2, and IFN-γ production. (B) Number of IFN-γ+ IL-2+ OT-I cells in the spleen. Data are shown as mean ± SEM. *, P < 0.01. (C) Granzyme B expression. Numbers below the granzyme B plots show mean fluorescence intensity (MFI). (D) CXCR3−/− or WT memory OT-I cells were generated in separate hosts and isolated from spleen at day 40+ after infection. CXCR3−/− and WT memory OT-I cells were CFSE labeled or not and adoptively transferred in a 1:1 mixture (1.9 × 106 cells total). (E) Recipient mice were rechallenged with VV-OVA 1 d after transfer. Representative plots showing proliferation of both OT-I cells on day 3 after infection as determined by CFSE dilution. Data are shown as mean ± SEM. (F) 1 d after transfer of unlabeled CXCR3−/− or WT memory OT-I cells, OVA peptide–pulsed (CFSEhi) and unpulsed (CFSElo) target cells were injected and killing was assessed 4 h later. Data are shown as mean ± SEM. (G) 1 d after transfer of unlabeled memory OT-I cells, recipient mice were challenged i.v. with 1 × 106 CFU LM-OVA. Bacterial numbers in the spleen were determined 3 d after challenge. Bars represent mean. *, P < 0.001; **, P < 0.005. Data represent three (A–C) or two (D–G) independent experiments (n = 3–4 per time point). (H and I) 1 × 104 naive splenic CXCR3−/− or WT OT-I cells were transferred into separate CD45.1 WT recipients, followed by i.v. infection with 2 × 106 PFU VV-OVA. After day 30+, the recipient mice were challenged i.v. with 1 × 107 CFU LM-OVA. Data represent two independent experiments (n = 4–6 per group). (H) Flow cytometry plots show the frequency of CXCR3−/− or WT OT-I cells in PBMC 1 d before LM-OVA challenge. (I) Bacterial titers in the spleen at 3 d after challenge. Bars represent mean. *, P < 0.05.