Localization of the meningococcal receptors for human transferrin
| Autor: | R.A. Wall, Dlawer A. A. Ala'Aldeen, S. P. Borriello, N. B. L. Powell |
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| Jazyk: | angličtina |
| Rok vydání: | 1993 |
| Předmět: |
Antigenicity
Immunology Cell Blotting Western Biology Neisseria meningitidis Microbiology Binding Competitive In vivo Iron-Binding Proteins Receptors Transferrin medicine Animals chemistry.chemical_classification Transferrin Iron-binding proteins Transferrin-Binding Proteins Molecular biology In vitro Blot Infectious Diseases medicine.anatomical_structure chemistry Parasitology Gold Rabbits Bacterial outer membrane Carrier Proteins Research Article Bacterial Outer Membrane Proteins |
| Popis: | The interaction between gold-labelled human transferrin (Au-HTF) with live meningococci after growth in vivo or in different in vitro conditions was examined by electron microscopy to localize and quantify the numbers of HTF-binding sites on the cell surface. It was clearly demonstrated that HTF binds to the surface of live meningococci (of different serogroups and serotypes) after growth in either iron-sufficient or iron-restricted cultures, although the degree of labelling was always higher (2- to 35-fold) in the latter case. The commensal Neisseria polysaccharea behaved similarly. Ultrathin sections showed that Au-HTF was localized predominantly on the outer membrane of the cells and vesicles, with hardly any internalization. Au-HTF labelling on meningococci was significantly reduced after incubation with unlabelled HTF or with rabbit antiserum containing antibodies against transferrin-binding proteins (TBPs), demonstrating the specificity of the interaction. These sera also blocked binding between HTF and outer membrane proteins on Western immunoblots. Direct evidence of the expression of the TBPs (Western blots) and localization of the HTF receptor (electron microscopy) on in vivo-grown meningococci was obtained from organisms derived without laboratory culturing from the cerebrospinal fluid of a patient. There was considerable cell-to-cell variation in the amount of labelling present on cells of the same sample (in vitro- or in vivo-grown organisms) and between different strains. The degree of binding varied with time of incubation of the cells with Au-HTF. The gold particles frequently formed discrete circles on the cell surfaces of the in vitro-grown organisms; these circles appear to be associated with outer membrane vesicle formation. The results show that the TBPs, which form part of the active components of the HTF receptor(s), are expressed in vivo and are surface exposed and immunogenic and that antibodies against them can interfere with the HTF binding of the meningococcal cells, which may affect iron utilization. This study further supports the concept of regarding the TBPs as future vaccine candidates. |
| Databáze: | OpenAIRE |
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