HS-AFM and SERS Analysis of Murine Norovirus Infection: Involvement of the Lipid Rafts

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Aybeke, Ece | Belliot, Gaël | Lemaire-Ewing, Stéphanie | Estienney, Marie | Lacroute, Yvon | Pothier, Pierre | Bourillot, Eric | Lesniewska, Eric

Edité par CCSD ; Wiley-VCH Verlag -

International audience. Studies on human norovirus are severely hampered by the absence of a cellculture system until the discovery of murine norovirus (MNV). The cell membranedomains called lipid rafts have been defined as a port of entry for viruses. Thisstudy is conducted to investigate murine norovirus binding on the mouse leukemicmonocyte macrophage cell line. Lipid raft related structures are extracted from cellsby detergent treatment resulting detergent-resistant membrane (DRMs) domains. Thereal-time polymerase chain reaction technique is performed to detect the viral genome,thereby the MNV binding on the DRMs. The interactions between MNV and DRMsare investigated by high-speed atomic force microscopy (HS-AFM) combined withsurface-enhanced Raman spectroscopy (SERS). The inoculation of the virus ontocells results in the aggregations of detergent-resistant membrane domains significantly.The characteristic Raman band of MNV is found in inoculated samples. To be surethat these results are originated from specific interactions between DRM and MNV,methyl-β-cyclo-dextrin (MβCD) is applied to disrupt lipid rafts. The MNV bindingon DRMs is precluded by the MβCD treatment. The cholesterols chains are defined asa key factor in the interactions between norovirus and DRMs. The authors concludethat the MNV binding involves the presence of DRMs and cholesterol dependent.

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