Comparison of inducible nitric oxide synthase expression in the brains of Listeria monocytogenes-Infected cattle, sheep, and goats and in macrophages stimulated in vitro

Abstract

The expression of inducible nitric oxide synthase (iNOS) was studied in the brains of cattle, sheep, and goat that succumbed to a natural infection with Listeria monocytogenes. The lesions in infected brains are characterized by microabscesses, perivascular cuffs, gliosis, glial nodules, and large areas of malacia. Using immunocytochemistry, we detected bacteria in microabscesses, particularly in sheep and goats, and in areas without signs of inflammation, but not in perivascular infiltrates. iNOS was expressed by macrophage (MΦ)-type cells of microabscesses and glial nodules but rarely by MΦ in areas of malacia, as determined by immunohistochemistry with iNOS- specific antibodies. iNOS was not detected in perivascular cuffs. Major histocompatibility complex class II molecules (MHC-II), another marker of cell activation, showed a different pattern of distribution. Perivascular cuffs contained high numbers of MHC-II-positive cells, including some with MΦ characteristics. Microabscesses in sheep and goats showed low expression of MHC-II, particularly in iNOS-expressing cells. In cattle, the expression of markers for activated or recruited phagocytes, the calcium-binding proteins S100A8 and S100A9 (formerly called MRP-8 and MRP-14, respectively), was largely restricted to cells showing weak or undetectable iNOS expression; iNOS-positive MΦ showed a low expression of S100A8 and S100A9. Thus, iNOS is expressed by a restricted subset of MΦ in listeric encephalitis. In cultured sheep and goat MΦ, a low proportion of cells expressed iNOS upon activation by L. monocytogenes and gamma interferon, resulting in nitrite generation at least 1 order of magnitude lower than that in similarly treated cattle MΦ. Since these species differences were much less obvious in vivo, it appears that the well-known species variation in iNOS expression by MΦ could reflect an in vitro phenomenon.