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. 2020 Jul 14;11:1474. doi: 10.3389/fimmu.2020.01474

Figure 3.

Figure 3

Nicotine stimulation induces micro RNA (miR)-181a-5p and miR-150-5p in CD8+ T cell with a naïve-memory phenotype. Isolated CD8+ T cells from peripheral blood of rheumatoid arthritis patients were cultured for 48 h in presence of aCD3 stimulation. Messenger RNA (mRNA) and miRs were measured by quantitative PCR. (A) A scoring system was constructed to differentiate between naïve-memory and effector phenotype. A higher than median expression of IL7R, FOXO1, BCL6, and TCF7 mRNA were rewarded with one point each. Samples with at least three points were considered to have a naïve-memory phenotype. (B) The fold change (FC) of miRs in samples with naïve-memory vs. effector phenotype. Cells were stimulated with aCD3 alone (gray circles) or with an addition of 10 μM nicotine (black circles). (C,D) PD-1 mRNA (C), FOXO1 mRNA (D), and IFN-γ secretion (E) in cells treated with 10 μM nicotine compared to cells only exposed to αCD3. P-values were calculated with Mann-Whitney U-test or Wilcoxon matched-pairs signed rank test. S, smoker; NS, non-smoker.