1887

Abstract

The human pathogen can cause atypical pneumonia through adherence to epithelial cells in the respiratory tract. The major immunogenic protein, P1, participates in the attachment of the bacteria to the host cells. To investigate the adhesion properties of P1, a recombinant protein (rP1-II) covering amino acids 1107–1518 of the P1 protein was produced. This protein inhibited the adhesion of to human HEp-2 cells, as visualized in a competitive-binding assay using immunofluorescence microscopy. Previous studies have shown that mAbs that recognize two epitopes in this region of P1 also reduce adhesion. Therefore, peptides covering these epitopes, of 8 and 13 aa, respectively, were synthesized to further investigate the adhesion region. None of these synthetic peptides reduced the binding of to the receptors on the host cells. Instead, 10 overlapping synthetic peptides covering the whole of rP1-II were evaluated in the competitive-binding assay using immunofluorescence microscopy. A reduction in the number of microcolonies was seen, which was confirmed for five peptides using a POLARstar OPTIMA reader to measure fluorescence intensity. The number of microcolonies adhering to the host cells was significantly reduced by these five peptides. Further investigations showed that inhibiting peptide 7 (amino acids 1347–1396) of the major adhesin protein P1 bound directly to host receptors, suggesting that the observed -inhibiting peptides occupied HEp-2 receptors, which are otherwise available for P1-mediated adhesion.

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2007-11-01
2020-08-07
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