Nicotine Inhibits Memory CTL Programming
Figure 7
Chronic exposure to nicotine does not change naïve CTL’s memory programming.
OT-I transgenic mice were given drinking water supplemented with nicotine at 200 µg/ml for 60 days [34], [37]–[39]. Naïve OT-I cells were purified from the nicotine treated OT-I mice or un-treated control mice and stimulated for 3 days with 3SI in the presence or absence of nicotine at 10 µM. Rapamycin was used with or without nicotine. Cells were then harvested for adoptive transfer at a concentration of 106 cells/mouse for memory differentiation in recipient B6 mice [22], [24], [25] (A–E) or direct examination (F–G). A–C. Analysis of memory CTLs 30 days after transfer. A. Comparison of memory CTLs in blood programmed in vitro by IL-12 from control or nicotine experienced OT-I mice. B. Comparison of memory CTLs in blood programmed under different treatments. All of the naïve OT-I CTLs were from nicotine treated donor mice. Data for “NC water experienced” in (A) are the same as “Control” in (B). C. CD62L expression on memory CTLs from blood in (B). D. Memory mice from (B) were challenged with LM-OVA, and protection was compared 3 days after challenge in spleen. E. Memory CTL secondary expansion in the blood 3 days after LM-OVA challenged (the same mice in D) mice. F–G. In vitro stimulated cells (for 3 days) were harvested to examine the production of functional molecules (F) or transcription factors and mTOR signaling molecules (G). Data from one experiment is shown, representative of two other experiments with similar results. Asterisks indicate statistical significance. *, P<0.05; **, P<0.01; ***, P<0.001, NS, not significant.
