ABSTRACT To investigate whether plasminogen may feature in scrapie infection, we inoculated plasminogen-deficient (Plg ^−/− ), heterozygous plasminogen-deficient (Plg ^+/− ), and wild-type (Plg ^+/+ ) mice by the intracerebral or intraperitoneal (i.p.) route with the RML scrapie strain and monitored the onset of neurological signs of disease, survival time, brain, and accumulation of scrapie disease-associated forms of the prion protein (PrP ^Sc ). Only after i.p. inoculation, a slight, although significant, difference in survival ( P < 0.05) between Plg ^−/− and Plg ^+/+ mice was observed. Neuropathological examination and Western blot analysis were carried out when the first signs of disease appeared in Plg ^+/+ animals (175 days after i.p. inoculation) and when mice reached the terminal stage of illness. At the onset of symptoms, PrP ^Sc accumulation was higher in the brain and spleen of Plg ^+/+ and Plg ^+/− mice than in those of Plg ^−/− mice, and these differences were paralleled by differences in the severity of spongiform changes and astrogliosis in the cerebral cortex and subcortical gray structures. Immunohistochemical analysis of the spleens before inoculation did not show any impairment of the immune system affecting follicular dendritic or lymphoid cells in Plg ^−/− mice. Once the disease progressed and mice began to die of infection, differences were no longer apparent in either brains or spleens. In conclusion, our data indicate that plasminogen has no major effect on the survival of scrapie agent-infected mice.