On the mountains, along an elevation gradient, we generally observe an ample variation in temperature, with the associated difference in vegetation structure and composition and soil properties. With the aim of quantifying the relative importance of temperature, vegetation and edaphic properties on soil respiration (SR), we investigated changes in SR along an elevation gradient (404 to 2101 m a.s.l) in the southern slopes of the Alps in Northern Italy. We also analysed soil physicochemical properties, including soil organic carbon (SOC) and nitrogen (N) stocks, fine root C and N, litter C and N, soil bulk densities and soil pH at five forest sites, and also stand structural properties, including vegetation height, age and basal area. Our results indicated that SR rates increased with temperature in all sites, and 55–76% of SR variability was explained by temperature. Annual cumulative SR, ranging between 0.65–1.40 kg C m-2 yr-1, decreased along the elevation gradient, while temperature sensitivity (Q10) of SR increased with elevation. However, a high SR rate (1.27 kg C m-2 yr-1) and low Q10 were recorded in the mature conifer forest stand at 1731 m a.s.l., characterized by an uneven-aged structure and high dominant tree height, resulting in a nonlinear relationship between elevation and temperature. Reference SR at 10°C (SRref) was unrelated to elevation, but was related to tree height. A significant negative linear relationship was found between bulk density and elevation. Conversely, SOC, root C and N stock, pH, and litter mass were best fitted by nonlinear relationships with elevation. However, these parameters were not significantly correlated with SR when the effect of temperature was removed (SRref). These results demonstrate that the main factor affecting SR in forest ecosystems along this Alpine elevation gradient is temperature, but its regulating role can be strongly influenced by site biological characteristics, particularly vegetation type and structure, affecting litter quality and microclimate. This study also confirms that high elevation sites are rich in SOC and more sensitive to climate change, being prone to high C losses as CO2. Furthermore, our data indicate a positive relationship between Q10 and dominant tree height, suggesting that mature forest ecosystems characterized by an uneven-age structure, high SRref and moderate Q10, may be more resilient.