engleski

The quantification of sialic acids released from guinea pig primary cell cultures for investigation of mumps virus entry and infection

Mumps virus (MuV), an aerosol-transmitted human pathogen, has two envelope glycoproteins, hemagglutinin-neuraminidase (HN) and a fusion (F) protein, which engage in receptor binding and mediate membrane fusion to the target cells. MuV- HN specifically recognize sialic acid (SA) containing structures of glycoconjugates present on the host cells, preferring unbranched α2, 3- sialylated glycans. SAs are negatively charged monosaccharides found on the non-reducing termini of glycans which are involved in many biological interactions. A diverse range of SAs are found in nature, but N-acetylneuraminic acid (Neu5Ac) and N-glycolylneuraminic acid (Neu5Gc) are the most frequent SAs in mammals. Guinea pigs are resistant to robust, symptoms- involving MuV infection. One of hypothesis is that this is due to lack of receptors for MuV on guinea pig cells, thus preventing virus entrance and infection. The aim of our work was to investigate whether cells isolated from guinea pig organs have properly glycosylated surface proteins, and whether the primary cultures prepared from these organs are susceptible to MuV infections. For this purpose we developed HPLC method for identification and quantification of fluorescently labelled SAs which are released by treatment guinea pigs primary cells with 2 different sialidases. The HPLC analysis showed the presence of both α2, 3- linked Neu5Ac and α2, 6-linked Neu5Ac on the surface of all analysed primary cell cultures. The α2, 3- linked Neu5Ac was more abundant in the all analyzed cells in comparison to α2, 6-linked Neu5Ac. In according to this results we detected the high susceptibility of the all analysed primary cells to infection with different mumps virus strain.

mumps virus, sialic acid, guinea pig primary cell cultures, sialidases, lectins, HPLC, flow citometry

nije evidentirano