We studied lichen communities along forest edge-to-interior gradients on opposite sides of a major highway and along a remote lakeshore in central North Carolina, U.S.A., to investigate highway pollution effects on this sensitive ecosystem component. At each site we sampled lichens on trees at 10 m intervals along each of five parallel transects established at the forest edge and at 25, 60, 100, and 150 m into the forest in the highway sites, with a similar layout along a nonlinear lakeshore in a similar forest type, from which transect distances from the forest edge were estimated using average tree distances from the nearest shoreline. Lichen communities were inventoried on tree trunks from the base up to 1.5 m height, then compared both among and within sites. Species richness was highest in the control site, and did not differ between the two highway sites. The highway sites were more similar to one another than either was to the control site, based on Bray-Curtis similarity indices. No associations were detected among sites and sampled lichen biotic components in terms of growth form, photobiont type or reproductive mode. In the highway sites, total transect species richness increased from the forest edge to 150 m distant. In the control site, species richness decreased from the forest edge to the most distant transect. Findings suggest a negative effect of highway pollution on species richness of lichen communities, but not on species composition by habit, photobiont type or reproductive mode. Despite the elevated NOx concentrations recorded along the highway, known nitrophilous species were largely absent, suggesting that other factors, including other pollutants, were affecting community structure near the highway.