Figure 6.
Intralymphatic CTL Transfer and Mathematical Modeling Show Low Killing Rates
Protocol for intralymphatic transfer: MCMV-3D-ΔvRAP footpad infection (0 hr), intralymphatic injection of different types of CTLs (∼4 hr), and two-photon imaging of single time points (∼24 hr).
(A) One day after infection and intralymphatic delivery of M45-tetramer-enriched CTLs (red) or M45-, M38-, and M139-tetramer-sorted CTLs (blue), the number of MCMV-3D-ΔvRAP-infected cells and CTLs per imaging region was counted from images of single time points.
(B) Same as (A) either without cell transfer or after injection of negatively enriched CTLs (magnetic depletion of CD62Lhi and non-CD8+ T cells).
(C) Same as (A) after injection of perforin-deficient tetramer-sorted CTLs.
(A–C) Data pooled from seven independent experiments (dots represent lymph nodes, and lines represent exponential decay). CTL priming was performed with intraperitoneal infection of CTL-donor mice with MCMV-3D.
(D) Mathematical modeling calculating the kinetics of infected cell numbers in a standard imaging region at the infected site of the lymph node cortical sinus, depending on the number of CTLs present (plot of infected cell numbers over time).
(E) The number of infected cells killed per T cell per day (PCKR) for the different CTL populations was calculated by the mathematical model from the raw data (median and 95% confidence interval) shown in (A)–(C). See Supplemental Experimental Procedures for details on the mathematical model.