Competitive inhibition and attachment assays in cell cultures to detect pathogenic binding components of mycoplasmas: a review.
A microattachment assay for quantitating adherence of radiolabelled Mycoplasma pneumoniae to human WiDr cell culture monolayers is described. Preincubating the WiDr cell monolayers with a protein-rich extract of M. pneumoniae inhibited the subsequent attachment of radiolabelled organisms. Competitive attachment inhibition provided a quantitative procedure to determine M. pneumoniae-binding components in the extract. The microattachment assays also measured attachment inhibition by the sialoglycoconjugates ceruloplasmin, orosomucoid and gangliosides, indicating that these reagents may be structural analogues of the mammalian cell receptor. Attachment of virulent M. pneumoniae strains to glutaraldehyde-treated monolayers was reduced approximately 60% and showed a different temperature dependence compared with untreated cells. These results suggest that maximal attachment of virulent M. pneumoniae may require two or more different receptors and binding components.