This study reports on the immunocytochemical localization of tumor necrosis factor-alpha (TNF alpha) in immune-mediated demyelination and Wallerian degeneration of the rat peripheral nervous system (PNS) using teased nerve fiber preparations. In experimental autoimmune neuritis induced by active immunization (EAN) or by adoptive transfer of autoreactive T cells (AT-EAN), macrophages passing blood vessels as well as macrophages adherent to nerve fibers were TNF alpha-positive. Large post-phagocytic macrophages at later stages of demyelination were TNF alpha-negative. Intraperitoneal application of an anti-TNF alpha antibody to EAN rats significantly reduced the degree of inflammatory demyelination, suggesting a pathogenic role for TNF alpha. After nerve transection only macrophages located within degenerating nerve fibers were TNF alpha-positive, while those entering and leaving nerves were negative. TNF alpha produced by macrophages seems to be involved in immune-mediated demyelination and non-immune myelin degradation after axotomy. While interferon-gamma (IFN gamma) is present in EAN nerves and may act as a local stimulus for TNF expression, the nature of this signal in Wallerian degeneration in the absence of IFN gamma is unknown.