Adherence of bacteria to mammalian cells: inhibition by tunicamycin and streptovirudin

Abstract

Group B streptococci were labeled either by growing the cells in ['4C]fructose or by using the surface label 4,4'-[3H]diisothiocyano-1,2-diphenylethane-2,2'-disulfonic acid, which reacts with amino groups. A quantitative assay was developed by using these labeled bacteria to study the adherence of streptococci to canine kidney epithelial celLs. The bacteria adhered to kidney celLs that had been infected with influenza A virus, but did not adhere to uninfected cells. The binding of 3Hlabeled group B streptococci was proportional to the number of bacteria added and showed saturation kinetics. The binding was blocked by the addition of unlabeled group B streptococci but was not affected by addition of streptococci from other groups. It was also blocked by mixing the 3H-labeled streptococci with influenza A virus before adding the bacteria to the kidney cells. When the kidney cells were infected with influenza virus in the presence of either tunicamycin or streptovirudin, these antibiotics inhibited the appearance of viral hemagglutinin in the kidney cells and also prevented the release of mature virus. In these experiments, the adherence of 'H-labeled streptococci was also inhibited. Tunicamycin was shown to block the incorporation of [14C]mannose into lipid-linked oligosaccharides and glycoprotein in both normal and virus-infected kidney cells. These data give strong support to the notion that adherence of streptococci to mammali cells involves recognition ofviral hemagglutinin, a glycoprotein whose synthesis is blocked by certain antibiotics.

Alterations in the Structure of the Oligosaccharide of Vesicular Stomatitis Virus G Protein by Swainsonine

“…These data correlate well with previous studies by Simonsen et al, who showed that , maturation of VSV was completed by 4.5 to 5.5 I h after the start of infection (34). Similar results cn were observed with influenza virus maturation 0 and the antibiotic tunicamycin (27).…”

Sialic acid-dependent interaction of group B streptococci with influenza virus-infected cells reveals a novel adherence and invasion mechanism

“…Both the co‐sedimentation of R1 with S. suis and of R2 with GBS were mediated by a sialic acid‐dependent interaction because both viruses remained in the supernatant when the respective streptococci were pretreated with neuraminidase (NA; Figure a). The NA pretreatment has been confirmed not to affect the bacteria viability (data not showed), which is consistent with previous reports (Butler, Baker, & Edwards, ; Pan, Schmitt, Sanford, & Elbein, ). A similar result was obtained when the supernatants were analysed by an infectivity assay (Figure b).…”

Adherence of staphylococcus aureus to influenza A virus-infected Madin-Darby canine kidney cell cultures

“…Isotopically labeled bacteria, group B Streptococcus type Ic and S. aureus (strains 1071 and P116), were used in a quantitative assay of bacterial binding to virus-infected and control MDCK cells. The streptococci were labeled with 4,4-[3H]diisothiocyano-1,2-diphenylethane-2,2'-disulfonic acid ([3H]DIDS) (specific activity, 2.024 x 109 cpm/.umol) as previously described (17). This compound is a surface label which forms covalent bonds with amino groups.…”