Assessing the Siginificance of Vertebrate Collagen and Fibronectin in Adhesion of Yersinia pseudotuberculosis and Yersinia pestis Using an Optical Trap

Currently, much attention is paid to the study of mechanisms of pathogen adhesion to human cells and tissues. Along with standard microbiological techniques, advanced biophysical methods are widely used for this purpose, among which a special place is occupied by optical trapping, which allows catching and moving individual bacterial cells with measurement of the interaction force between them and target molecules. The aim of this work was to evaluate the significance of collagen and fibronectin in adhesion of Yersinia pseudotuberculosis O1b and the vaccine strain Yersinia pestis EV NIIEG, cultivated at different temperatures, applying optical trap. Materials and methods. Bacteria grown at two temperatures (+10 and +37 °C for Y. pseudotuberculosis or +27 and +37 °C for Y. pestis) were caught by a laser beam and stepwise brought to the protein-coated glass. One second after the contact, the cell was retracted in semi-automatic mode at a constant speed until a spike was observed on the signal chronogram, and its voltage then was converted into force units. Differences between data sets were determined by the mean and median forces, as well as by the results of constructing histograms of the distribution of interaction forces. Results and discussion. The significance of collagen in the adhesion of Y. pseudotuberculosis grown at +37 °C but not at +10 °C was shown. Fibronectin demonstrated no pronounced differences between the bacteria of the two Yersinia species at all the cultivation temperature modes. In all cases, affinity of Yersinia cells with collagen and fibronectin was higher compared to the control, bovine serum albumin: 7.2, 8.1, and 2.0 pN for 1b-10 cells; 12.4, 7.6, and 4.7 pN for 1b-37 cells; 6.1, 6.6, and 4.4 pN for EV-27 cells; 7.4, 7.3, and 4.3 pN for EV-37 cells, respectively. The binding of Yersinia to collagen and fibronectin is probably due to the physicochemical properties of the bacterial surface structures and components of connective tissue, as well as experimental conditions. Revealing the mechanisms of such interactions requires additional investigations of individual Yersinia antigens applied onto polystyrene beads. The method may be introduced when working with other pathogens too.

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