Figure 3

Correlation of T Cell Dose and Luciferase Bioluminescent Signal. A comparison of T-lux T cell numbers with light emission signal was done both in vitro and in vivo to assess the sensitivity of the T cell luciferase signal. (A) Pooled CD3⁺ T cells from spleen and peripheral lymph nodes of T-lux mice were plated in triplicate (in columns) into a 96-well plate at the indicated cell numbers per well, and light emission was quantitated following addition of luciferin. Pseudocolor scale is shown to right. (B) Plot of light emission data from (A) versus input T cell numbers. Error bars indicate standard error of the mean (SEM). Medium alone was used as negative control. The r² value for data is displayed on figure (see text for details). (C) CD3⁺ T cells isolated from pooled spleen and LN of T-lux mice were adoptively transferred into congenic C57BL/6 recipients. Whole body bioluminescence was quantified at 12, 24, and 72 hours post-transfer. Representative images from 72 hours post-transfer are shown for three cell doses. (D) Whole body luminescence was plotted against transferred T cell numbers in (C). A representative plot is shown for the 72-hour time point.