Profita_2014_Immunobiology_219_392

T-lymphocytes, including Th17-cells and T-cells expressing acetylcholine (ACh), are key components of systemic inflammation in chronic obstructive pulmonary disease (COPD). We investigated whether ACh promotes Th17 cells in COPD. ACh, IL-17A, IL-22, RORgammat, FOXP3 expression and AChIL-17A, AChIL-22, AChRORgammat coexpression was evaluated in peripheral blood mononuclear cells (PBMC) from COPD patients (n=16), healthy smokers (HS) (n=12) and healthy control subjects (HC) (n=13) (cultured for 48h with PMA) by flow cytometry. Furthermore, we studied the effect of Tiotropium (Spiriva((R))) (100nM) and Olodaterol (1nM) alone or in combination, and of hemicholinium-3 (50muM) on AChIL-17A, AChIL-22, AChRORgammat, and FOXP3 expression in CD3(+)PBT-cells of PBMC from COPD patients (n=6) cultured for 48h with PMA. CD3(+)PBT-cells expressing ACh, IL-17A, IL-22 and RORgammat together with CD3(+)PBT-cells co-expressing AChIL-17A, AChIL-22 and AChRORgammat were significantly increased in COPD patients compared to HC and HS subjects with higher levels in HS than in HC without a significant difference. CD3(+)FOXP3(+)PBT-cells were increased in HS than in HC and COPD. Tiotropium and Olodaterol reduced the percentage of CD3(+)PBT-cells co-expressing AChIL-17A, AChIL-22, and AChRORgammat while increased the CD3(+)FOXP3(+)PBT-cells in PBMC from COPD patients, cultured in vitro for 48h, with an additive effect when used in combination. Hemicholnium-3 reduced the percentage of ACh(+)IL-17A(+), ACh(+)IL-22(+), and ACh(+)RORgammat(+) while it did not affect FOXP3(+) expression in CD3(+)PBT-cells from cultured PBMC from COPD patients. We concluded that ACh might promote the increased levels of Th17-cells in systemic inflammation of COPD. Long-acting beta2-agonists and anticholinergic drugs might contribute to control this event.