Macrophages play an important role in asthma pathogenesis both in the inflammatory and resolution phase of the disease. Macrophages can acquire different polarisation states dependent on their microenvironment. It is yet unclear through which mechanism the microenvironment affects the anti-viral response in macrophages. We hypothesized that the macrophage microenvironment regulates rhinovirus-induced IFNbeta expression. Murine bone marrow-derived monocytes and human differentiated THP-1 cells were stimulated with M-CSF or GM-CSF and IFNgamma or IL-4/IL-13, respectively, to mimic a Th1 or Th2 environment. Macrophages were infected with rhinovirus and gene and protein levels of IFNbeta and pattern recognition receptor expression were measured. In subsequent experiments an IkappaB kinase inhibitor was used to study the involvement of NFkappaB. Both murine and human M1-like macrophages exhibited higher levels of IFNbeta and pattern recognition receptors after rhinovirus infection than M2-like macrophages. Blockage of NFkappaB resulted in a lower expression of rhinovirus-induced IFNbeta in human M1-like macrophages while inducing a higher expression in M2-like macrophages, suggesting that the interferon response towards viral infection was mediated by NFkappaB. These findings could contribute to a better understanding of mechanisms causing reduced anti-viral responses at viral-induced exacerbations in asthma.