<i>Objective:</i> Smoking is generally accepted as a factor that affects the disease course in inflammatory bowel disease patients. Whether these effects can be contributed to the immunomodulatory effects of nicotine via nicotinic acetylcholine receptor (nAChR) activation is unclear. As previous data suggest that the α7 nicotinic acetylcholine receptor <i>(CHRNA7)</i> and its duplicated variant <i>CHRFAM7A</i> may specifically participate in the inflammatory response of monocytes, we evaluated whether repeated nicotine exposure or smoking affects monocyte <i>CHRNA7 </i>and<i> CHRFAM7A</i> expression and cholinergic immunomodulation. <i>Methods:</i> The human monocyte cell line THP-I was incubated with nicotine for different time points before endotoxin exposure. In a pilot volunteer study using smoking (n = 4) and nonsmoking (n = 7) individuals, vagal output was stimulated by olive oil administration after which monocytes were analyzed for nicotinic receptor expression. Serum tumor necrosis factor (TNF) levels were determined using ELISA and expression levels of the nAChR subunits <i>CHRNA7</i>, <i>CHRNB2</i> or <i>CHRFAM7A </i>were analyzed using QPCR. <i>Results:</i> Repeated nicotine exposure upregulated <i>CHRNA7</i> expression on THP-I monocytes and led to an enhanced potential of α7 nAChR agonist GSK1345038A to reduce TNF levels. Furthermore, <i>CHRNA7</i> was only detectable in isolated blood monocytes of smokers. On the other hand, the expression of <i>CHRFAM7A</i> and <i>CHRNB2</i> was not affected by nicotine exposure. Lipopolysaccharides-induced TNF secretion was inhibited by nicotinic receptor activation in THP-I monocytes, but this response was not consistently seen in blood monocytes from smoking individuals. <i>Conclusions:</i> We conclude that <i>CHRNA7 </i>expression on blood monocytes is upregulated in smoking individuals, which may contribute to cholinergic immunomodulation.