T cell immunity and interferon-γ secretion during experimental allergic encephalomyelitis in Lewis rats

Abstract

An immunospot assay that detects single secretory cells was used to enumerate interferon-γ secreting cells (IFN-γ-sc) in mononuclear cell suspensions from the central nervous system (CNS) and peripheral lymphoid organs after actively induced experimental allergic encephalomyelitis (EAE) in Lewis rats. In the CNS compartment there was a significant increase in the number of IFN-γ-sc preceding the onset of the clinical signs of EAE. Both in rats with EAE and rats immunized with Freund's complete adjuvant (FCA) the number of IFN-γ-sc increased in peripheral lymphoid organs, as compared to non-immunized controls. In view of the potent immunoregulatory effects of IFN-γ, its intra-CNS secretion may play a crucial role for clinicophatological events in EAE. To study the numbers of primed T cells that in response to myelin antigens produced IFN-γ, mononuclear cell suspensions from peripheral lymphoid organs were precultured to allow for antigen uptake, presentation and T cell triggering, followed by enumeration of IFN-γ-sc. T cells responding to a peptide of myelin basic protein (MBP) that previously have been shown encephalitogenic in Lewis rats, appeared initially and were quantitatively dominant over the course of EAE. Later, T cell reactivities to multiple regions of MBP appeared, showing that the concept of immunodominance in EAE is non-absolute and time dependent. Splenocyte cultures from EAE rats exposed to the different antigens showed a reduced number of IFN-γ-sc compared to cultures not exposed to antigen, suggesting an antigwn-induced suppression of T cell effector molecules.