Cell “Flakes” in Winter

The objective of my study is to gain an understanding of the molecular events in Theiler’s murine encephalomyelitis virus (TMEV) infection resulting in cell death. TMEV is a highly cytolytic RNA virus that causes a persistent central nervous system (CNS) infection and immune-mediated demyelination in susceptible strains of mice, providing a relevant experimental animal model for multiple sclerosis (MS). It has been shown that infection with TMEV induces apoptosis in murine macrophages, resulting in restricted infectious virus production. In contrast, BeAn infection in all other rodent cells (e.g Syrian baby hamster kidney cells (BHK-21)) induces necrotic cell death with high virus yields. Currently the cellular factors essential in determining the fate of TMEV-infected cells are largely unknown. To determine whether members of the Bcl-2 family act as key anti-apoptotic proteins that regulate and determine the mechanism of TMEV-induced cell death, I transiently transfected BHK-21 cells with siRNA oligos to knock-down the expression of different anti-apoptotic proteins and infected these cells with TMEV virus. As a negative control, I used mock infected BHK-21 cells. To visualize apoptotic profile under the fluorescent microscope, I stained the cells with a nucleic acid specific dye, 4'-6-Diamidino-2-phenylindole (DAPI). The fluorescence image shows mock-infected BHK-21 cells grown in cover slips that were stained with DAPI.