On-highway tractor-trailer vehicles operate in a complex aerodynamic environment that includes influences of surrounding vehicles. Typical aerodynamic analyses and testing of single vehicles on test track, in wind tunnel or in computational fluid dynamics (CFD) do not account for these real world effects. However, it is possible with simulation and on-road testing to evaluate these aerodynamic interactions. CFD and physical testing of multiple vehicle interactions show that traffic interactions can impact the overall drag of leading and trailing vehicles. This paper will discuss results found in evaluating the effects of separation distances on tractor-trailer aerodynamics in on-road and CFD evaluations using a time-accurate Lattice Boltzmann Method based approach and the ramifications for improving real world prediction versus controlled single vehicle testing.