Simplified quantitative analysis of spinal cord cells from Theiler's virus-infected mice without the requirement for myelin debris removal.

Flow cytometry is routinely used to analyze mononuclear cell populations in experimental CNS infections and autoimmune diseases in mice and other rodents. Previous analysis of mononuclear cell samples has involved centrifugation of single cell suspensions from minced CNS tissues on discontinuous gradients to separate cells from myelin debris: however, loss of cells that occurs on gradients necessitates pooling of CNS tissues. We have developed a method of analyzing cells in CNS tissues by flow cytometry without removing myelin debris, and applied this method to assess mononuclear cells and oligodendrocytes in SJL mice with Theiler's murine encephalomyelitis virus (TMEV)-induced demyelinating disease.

Apoptotic cells, including macrophages, are prominent in Theiler's virus-induced inflammatory, demyelinating lesions.

Theiler's murine encephalomyelitis virus (TMEV) persists in the mouse central nervous system principally in macrophages, and infected macrophages in culture undergo apoptosis. We have detected abundant apoptotic cells in perivascular cuffs and inflammatory, demyelinating lesions of SJL mice chronically infected with TMEV. T cells comprised 74% of apoptotic cells, while 8% were macrophages, 0.