The ambient PM2.5-related health burden (all-cause and cause-specific mortality, and years of life lost (YLL)) and its economic impacts in Isfahan during the period 2016–2019 were assessed. Ambient PM2.5-related premature deaths were estimated using the Environmental Benefits Mapping and Analysis Program based on the most common global function, called the Global Exposure Mortality Model. The annual PM2.5 concentrations in Isfahan was measured to be approximately six times higher (27.8–32.1 μg m−3) than the recommended by the World Health Organization value (5 μg/m3). Ambient PM2.5-related all-cause mortality in Isfahan was in the range of 1311–1456 over the four-year period. Moreover, approximately 15% of all-cause mortality in adults (age 25 and above) was attributed to ambient PM2.5 in Isfahan. Ischemic heart disease and stroke caused about 63% of PM2.5-related total deaths in the city. The YLL attributable to ambient PM2.5 exposures ranged from 22,488 to 25,614. The results demonstrated that avoided annual economic costs associated with reducing PM2.5 ranged from $526,008,055 to $599,945,966 over the study period. The corresponding results for mortality, morbidity, and economic results would provide a measurable assessment to the policymakers of the city of Isfahan in order to intensify their efforts on improvement the ambient air quality.