In-service Portland cement concrete (PCC) pavements are subject to repeated dynamic loads from moving vehicles; thus, the actual stress generated in a PCC pavement may significantly differ from the static stress, which is normally used in the design and evaluation of pavement performance. Calculating the stress in PCC pavements under moving vehicle loads is of importance to assess their actual service condition, particularly for pavements with different surface roughness levels as the deteriorated roughness might cause large stress in PCC pavement subject to dynamic loads. In this paper, a method is proposed to compute the dynamic response in terms of loads and stresses generated in jointed plain concrete pavements (JPCPs) under a moving axle load, considering the effects of the pavement surface roughness, the vehicle parameters (including vehicle speeds and axle weights), and the pavement structure parameters (including thickness and elastic modulus of different layers and the existence of dowel bars). The dynamic axle load is firstly generated based on the quarter-car model, running through three successive slabs of which the surface roughness is determined by the power spectral density method, and the critical locations in slabs where the largest tensile stresses occur are identified. The combined effects of various pavement surface roughness levels, vehicle speeds, axle weights, and pavement structure parameters are evaluated in terms of the stress and the dynamic factor defined as the ratio of the tensile stress under dynamic load to the tensile stress under static load. For the roughness level D, the tensile stress can reach a maximum value of 3.13 MPa, and the dynamic factor can reach a maximum value of 2.46, which is much larger than the dynamic factor of 1.15 or 1.2 currently used in design guidebooks. Increasing the thicknesses of pavement slab or the subbase layer is an effective way to reduce the tensile stress in JPCP, while increasing the thickness of base layer is not effective. The results of this study can benefit future pavement design and pavement performance evaluation by providing the actual stress and the useful dynamic factor values for various conditions of field pavements. Moreover, preventive maintenance, particularly the improvement of pavement surface roughness, can be planned by referring to the results of this study, to avoid large tensile stress generated in JPCPs.