Updated project metadata. ASFV poses a continuing threat to the pig industry in Europe as broadly applicable vaccines are not available. Variations within the ASFV genome resulted in the emergence of attenuated strains with low or moderate virulence, however, the molecular basis of the differences in virulence is not known. To reveal virulence-associated protein expression patterns, we have analyzed the proteomes of the natural target cells of ASFV, primary porcine macrophages, after infection with two genotype II ASFV strains displaying high (“Armenia 2008”) and moderate (”Estonia 2014”) virulence using quantitative mass spectrometry. Very similar expression patterns were observed for the viral genes. In addition to the canonical ASFV proteins, twelve novel protein products from recently described transcripts could be confirmed in both isolates. Pathway analysis showed that both isolates evoked a similar host proteome response despite their difference in virulence. However, subtle differences in the manipulation of the proteins involved in the proinflammatory response mediated by the MAPK14/p38 signalling cascade were observed.