To characterize the distribution and anisotropy of dislocations in cold-drawn pearlitic steel wires, X-ray diffraction line-profile analysis was performed using synchrotron radiation micro-beams. An analytical procedure for correcting the instrumental line broadening for highly directional micro-beams was developed using diffraction profiles of standard CeO2 powder. Although the CeO2 powder line profile includes line broadening due to its microstructural imperfections, the instrumental broadening can be obtained by estimating the effect of the microstructural imperfections on the line broadening. The plastic shear strain was generally more severe near the surface than the center of the wire, whereas the dislocation density distribution was almost constant from the center to the surface. On the other hand, the dislocation rearrangement, which evolves the dislocation cell structure, progressed closer to the surface. It was also revealed that a difference between the hardness in axial and transverse wire directions could be explained by anisotropic dislocation density. Line-profile analysis based on diffraction data at elevated temperatures was performed. Whereas the cementite recovery progressed at a constant rate, the ferrite phase recovery rate was temperature-dependent, suggesting that the ferrite phase recovery was less related to that of the cementite phase.