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Nave CD4 + T cells (0.4 10 6 ) were cultured in RPMI-1640 medium supplemented with 10% FBS (Gibco, CA, USA), 2 mM glutamine, 1% penicillin streptomycin (Gibco, CA, USA), 10 mmol/L N-2-hydroxyethylpiperazine-N-2-ethanesulfonic acid (HEPES, Gibco, CA, USA), and 55 mmol/L 2-mercaptoethanol (Gibco, CA, USA) in 24-well plates

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Due to its properties and effects, nicotine is classified as a substance of very high addiction liability