brivudine and 5-propyl-2--deoxyuridine

In herpes simplex virus-infected (HSV) cells, the antiviral nucleoside analogue 5-n-propyl-2'-deoxyuridine (PdU) may, under certain circumstances, induce a pattern of interference with late steps in formation of N-linked glycans, resulting in increased availability of viral glycoproteins for neutralizing antibodies. The PdU-induced changes in N-linked glycans, released by pronase digestion of the HSV-specified glycoprotein gC-1, were investigated by using lectin affinity chromatography and Bio-Gel P6 gel filtration of glycans, radiolabelled with [3H]galactose or [3H]glucosamine. PdU-treatment of HSV-infected cells totally inhibited addition of sialic acid and reduced the amount of galactose incorporated into N-linked glycans by 70%. In addition, the PDU-treatment caused a decrease in oligosaccharides with affinity for Phaseoulus vulgaris leuco-agglutinin and erythro-agglutinin, and an increase in Lens culinaris lectin (LCA)-binding oligosaccharides, suggesting a PdU-induced shift from multi-branched to moderately branched structures. This shift was also found in HSV-infected B16 mouse melanoma cells, where the large content of multi-branched oligosaccharides contributes to the metastatic potential. The LCA-binding glycans from PdU-treated cells were smaller and contained less galactose units than corresponding structures from untreated cells. In a cell-free system, PdU 5'-monophosphate inhibited the translocation of UDP-GlcNAc, and, to a smaller extent, also the translocation of UDP-galactose into Golgi vesicles, suggesting that nucleotide sugar translocation is one important target for the PdU-induced interference with glycosylation in HSV-infected cells.

Topics: Agglutinins; Animals; Antiviral Agents; Biological Transport; Bromodeoxyuridine; Cell Line; Chlorocebus aethiops; Chromatography, Affinity; Chromatography, Gel; Deoxyuridine; Glycosylation; Golgi Apparatus; Humans; Infant; Lectins; Melanoma, Experimental; Mice; Neuraminidase; Nucleoside Diphosphate Sugars; Polysaccharides; Simplexvirus; Tumor Cells, Cultured; Viral Envelope Proteins

A rapid, reproducible and objective new method for typing herpes simplex viruses type 1 (HSV-1) and type 2 (HSV-2) based on the effects of virus-induced thymidine kinases on various antiviral drugs has been developed. When several laboratory strains and clinical isolates were typed by this method and compared to the results obtained by the immunofluorescence antibody typing method, agreement was found for all the viruses. The new technique has the added advantage of determining the sensitivity of HSV strains to antiviral drugs.

Topics: Bromodeoxyuridine; Cells, Cultured; Deoxyribonucleases; Deoxyuridine; Enzyme Induction; Fluorescent Antibody Technique; HeLa Cells; Humans; Idoxuridine; Microbial Sensitivity Tests; Simplexvirus; Thymidine Kinase