Asthma is a chronic airway disease whose exacerbations are often triggered by rhinovirus infection. TGF-beta1 induces rhinovirus replication in infected cells. Moreover, TGF-beta1 is a pleiotropic mediator that is produced by many immune cells in the latent, inactive form bound to the latency-associated peptide (LAP) and to the transmembrane protein glycoprotein A repetitions predominant (GARP). In this study we wanted to investigate the effect of rhinovirus infection on the TGF-beta secretion and the downstream signaling via TGF-betaRI/RII in peripheral blood mononuclear cells from control and asthmatic patients after rhinovirus infection ex vivo. Here, we found a significant upregulation of TGF-betaRII in untouched PBMCs of asthmatics as well as a suppression of TGF-beta release in the rhinovirus-infected PBMC condition. Moreover, consistent with an effect of TGF-beta on Tregs, PBMCs infected with RV induced Tregs, and TGF-betaRII directly correlated with RV1b mRNA. Finally, we found via flow cytometry that NK cells expressed less GARP surface-bound TGF-beta, while cytokine-producing NK(bright) cells were induced. In summary, we show that rhinovirus infection inhibits TGF-beta release in PBMCs, which results in the activation of both Treg and NK cells.