We compared the short-term magnetic resonance imaging (MRI) evolution of new multiple sclerosis (MS) lesions enhancing after single dose (SD) (0.1 mmol/kg) or triple dose (TD) (0.3 mmol/kg) gadolinium-DTPA (Gd) to explore possible differences in the pathological substrates of acute MS lesions. Brain MRI scans were obtained at baseline and every 4 weeks for a 3-month period in 18 relapsing-remitting MS patients. At each time point, using two separate sessions, we obtained dual echo and T1-weighted scans before and after SD and TD of Gd. New enhancing lesions detected at month 1 and 2 were entered into the analysis. The presence of corresponding hypointense lesions on unenhanced T1-weighted scans and hyperintense lesions on T2/proton density (PD)-weighted images was assessed on the same scan and on the scans performed 1 month before and 1 month after the new lesion development. Persistence of enhancement was evaluated on the SD and TD scans obtained 1 month after new lesion appearance. One-hundred and sixty lesions were studied. Of these, 97 lesions were enhancing after both SD and TD (group A) and 63 lesions only after TD (group B). Thirty (31%) of the lesions enhancing after both SD and TD and ten (16%) of the lesions enhancing only after TD had corresponding T1-weighted lesions (P = 0.03). Of these lesions, 87% in group A and 40% in group B (P = 0.003) were not hypointense on the previous scans. No differences were found in the frequencies of corresponding T2/PD-weighted abnormalities (92% in Group A vs. 87% in Group B lesions). Of these hyperintense areas, 62% in group A and 56% in group B were not present on the previous scans. On follow-up scans, 52% of the lesions enhancing after SD and TD and 70% of the lesions enhancing only after TD did not show enhancement after the injection of both the doses of Gd (P = 0.02). The frequencies of corresponding T2/PD and T1-weighted abnormalities were higher in Group A than in Group B lesions, but the differences were not statistically significant. Our findings suggest that the pathological process is less severe in MS lesions enhancing only after TD injection than in those enhancing after the SD.