Figure 1
From: Lung B cells promote early pathogen dissemination and hasten death from inhalation anthrax
Labeled Bacillus anthracis (BA) spores are transported to the lung-draining lymph nodes (LDLNs) early after intratracheal (i.t.) infection. (a) Representative histogram demonstrating uniform labeling of BA spores with DyLight 649. Data were collected on an LSRII. Histogram shows labeled BA spores (black) compared with unlabeled BA spores (solid gray). DyLight 649-labeled spores were always checked for uniform labeling before using for an experiment. (b) Mice were infected i.t. with 1 × 106, 1 × 107, or 1 × 108 BA spores or given phosphate-buffered saline (PBS) alone. LDLNs were analyzed by fluorescence-activated cell sorting (FACS) at 6 h.p.i. Fluorescence from BA spores is plotted against an empty channel to show percentage of spore+ cells in the LDLNs. (c) LDLNs of infected A/J mice were analyzed for spore+ cells by FACS and plated for heat-resistant colony-forming units (CFUs). (d) A/J mice were given 1 × 108 BA spores i.t. and single-cell suspensions from LDLNs were analyzed by flow cytometry at 30 min, and 2 and 5 h after infection for total number of spore+ cells. Data in panels c are pooled from three independent experiments with n=2–3 per group. *P<0.05. Experiments in panels b and d are representative of two independent experiments with n=2–3 each.
