Role of lipid rafts in porcine reproductive and respiratory syndrome virus infection in MARC-145 cells

Lipid rafts play an important role in the life cycle of many viruses. Cholesterol is a critical structural component of lipid rafts. Although the porcine reproductive and respiratory syndrome virus (PRRSV) has restricted cell tropism for cells of the monocyte/macrophage lineage, a non-macrophage cell MARC-145 was susceptible to PRRSV because of the expression of virus receptor CD163 on the cell surface, therefore MARC-145 cells is used as model cell for PRRSV studies. In order to determine if Cholesterol is involved in PRRSV Infection in MARC-145 cells, we used three pharmacological agents: methyl-β cyclodextrin (MβCD), mevinolin, and filipin complex to deplete Cholesterol in MARC-145. Although these agents act by different mechanisms, they all significantly inhibited PRRSV Infection. The inhibition could be prevented by addition of exogenous Cholesterol. Cell membrane Cholesterol depletion after virus Infection had no effect on PRRSV production and Cholesterol depletion pre-infection did not reduce the virus attachment, suggesting Cholesterol is involved in virus entry. Further results showed that Cholesterol depletion did not change expression levels of the PRRSV receptor CD163 in MARC-145, had no effect on clathrin-mediated endocytosis, but disturbed lipid-raft-dependent endocytosis. Collectively, these studies suggest that Cholesterol is critical for PRRSV entry, which is likely to be mediated by a lipid-raft-dependent pathway.