In vitro induction of nitric oxide synthase in astrocytes and microglia byTrypanosoma brucei brucei

In stage II human african trypanosomiasis (HAT), which is characterized by central nervous system (CNS) involvement, neurones and oligodendrocytes mig ht be targets of dysimmune processes. Nitric oxide (NO) production by perip heral macrophages is documented in HAT. We studied the production of NO by murine astrocytes and microglia cocultured with Trypanosoma brucei (T. b.) brucei AnTat 1.9. Purified astrocytes or microglia from mouse brains were c ocultured with T. b. brucei, and in some instances with interferon (IFN)-ga mma, which is known to be released during the disease and also to be a grow th factor for trypanosomes. Inducible NO synthase (iNOS) expression was stu died by indirect immunofluorescence and reverse transcriptase-polymerase ch ain reaction. NO production was determined by measuring nitrite generation in culture. Detection of iNOS in astrocytes and microglia in the presence o f T. b. brucei, was closely associated with nitrite production and was stro ngly enhanced by the addition of IFN-gamma to the culture medium. The stimu lation of iNOS activity required parasite-cell contact and likely occurred at the transcriptional level. This study demonstrates the induction of iNOS in CNS-related macrophage cells in the presence of trypanosomes and its po tentiation by IFN-gamma.