The protozoan Neospora caninum causes a severe transplacental acquired neurologic disease in dogs and abortion in cattle with economic losses worldwide. Definitive hosts of N. caninum are dogs and coyotes (Canis latrans) that can shed oocysts. It is still unclear whether N. caninum is a human pathogen, once only a few cases with serological evidence suggestive of infection were reported, but in the absence of clinical symptoms. Studies in mice and cattle have shown that immunity to N. caninum infection involves predominantly Th1-type response. However, several studies on the immune system during pregnancy have demonstrated that this is biased towards a Th2-type response rather than from a Th1 response. Little is known about the immune response of the central nervous system against this protozoan. Recently, a studied reported that high levels of IL-10 were produced by infected astrocytes in vitro. Furthermore, TNF α and IFN γ suppressed the parasite growth in cerebellar cell cultures. The aim of this study was to evaluate the production of IL-6, IL-10, TNF α, IFN γ and NO in glial cell cultures infected by Neospora caninum. Astrocytes were obtained from the brain cortex of newborn rats (<48h of age). Cultures were maintained in DMEM supplemented with 10% fetal bovine serum, 1mM pyruvic acid and 2mM glutamine that was changed three times a week. Parasites were counted and a cell/parasite ratio of 1:1 was used to infect primary cell cultures. Cytokines were measured in the supernatant of control and infected cultures by using an ELISA commercial kit, and nitrite was used as a marker for NO production according to Griess method. The release of IFN γ observed in the culture medium of infected cultures did not differ significantly from the basal value measured in control conditions. Whereas control cells released only low levels of all cytokines, the mean levels released in the medium of infected cultures after 24 and 72h were respectively 3.8 ± 0.6 and 3.7 ± 0.6 pg /mg of proteins of TNF α, 2.7 ± 0.69 and 4.1 ± 0.64 pg/mg of protein of IL-10 and more than 4000 pg/ml of IL-6. NO level also increased 24h (2.3 ± 0.8 pg/mg de protein) post-infection and continued to increase until 72h (4.2 ± 1.1 pg/mg de protein). Together, these results suggest that glial cell cultures of rats present high levels of Th2-type cytokines, probably to suppress the harmful effects of IFN γ. Furthermore TNF α and NO should represent an effective response against N. caninum.