In laser additive manufacturing (AM) of hot work tool steels, direct tempering (DT) of the tool from as-built (AB) condition without prior conventional austenitization and quenching results in enhanced tempering resistance. To date, intercellular retained austenite (RA) decomposition, leading to a shift in secondary hardening peak temperature, and finer martensite substructure are reported to be responsible for such a behavior. In this work, authors aimed at studying the strengthening contributions by performing isothermal tempering tests for long times (up to 40 hours) at elevated temperatures (up to 650 degrees C) on DT and quenched and tempered (QT) specimens. The thermal softening kinetics and the microstructural evolution were evaluated with the support of computational thermodynamics. The results suggest that the main contributor to enhanced temper resistance in DT condition is the larger fraction of thermally stable and extremely fine (similar to 20 nm) secondary (tempering) V(C,N) compared with QT. This could be explained by the reduction of available V and C in austenitized and quenched martensite for a later secondary V(C,N) precipitation during tempering, because of equilibrium precipitation of relatively large (up to 500 nm) vanadium-rich carbonitrides during the austenitization process. A complementary effect of the substructure refinement (i.e., martensite block width) in rapidly solidified highly supersaturated martensite was also quantified in terms of Hall-Petch strengthening mechanism. The significant effect of secondary V(C,N) was successfully validated by assessing a laser AM processed vanadium-free hot work tool steel in QT and DT condition, where no significant differences in strength and temper resistance between the two conditions were evident. [GRAPHICS] .