Figure 5.

CTLs Show Low Average PCKRs in Both Poxvirus and Cytomegalovirus Infection Models

Experimental setup for two-photon imaging of MVA-infected lymph nodes: transfer of FP-OT-I (day 0), expansion with SIINFEKL plus poly(I:C) (day 1), infection with MVA or MVA-OVA (day 6), and two-photon imaging (day 7).

(A) One day after footpad infection with MVA or MVA-OVA (10⁷ PFU), two-photon microscopy was used for observing OT-I CTLs (green) at the site of MVA-infected mCherry⁺ cells (red).

(B) Median track speed of the OT-I CTL population in non-infected or MVA- or MVA-OVA-infected lymph nodes. Dots represent the mean of >50 CTLs analyzed per lymph node, and red bars represent the mean ± SD. A t test with Welch’s correction was used for comparing MVA and MVA-OVA. ^∗∗p < 0.01.

(C) OT-I CTL contacts with MVA- or MVA-OVA-infected cells were analyzed. Dots represent contact duration per CTL, and red bars represent IQRs. A Mann-Whitney test was used for comparing MVA and MVA-OVA. ^∗∗p < 0.01.

(D) One day after infection, the number of OT-I CTLs and MVA-infected cells per imaging region was correlated (green dots, MVA infection; green line, linear regression; red dots, MVA-OVA infection; red line, one-phase exponential decay). Dots represent individual mice (B), cells (C), and lymph nodes (D). Data were pooled from three independent experiments in (B)–(D). Scale bars represent 20 μm.

(E) OT-I CTLs PCKRs were calculated from automated-cell-tracking data for different MCMV and MVA strains. Dots represent movies, and red bars represent IQRs. A Kruskal-Wallis test was used for comparing multiple groups (outliers are not shown but were included in the test). ^∗p < 0.05; ns, not significant.