Adherence of Lactobacillus salivarius to HeLa Cells Promotes Changes in the Expression of the Genes Involved in Biosynthesis of Their Ligands

Abstract

The attachment of a variety of Lactobacilli to the mucosal surfaces is accomplished through the interaction of OppA, a superficial bacterial protein also involved in oligopeptide internalization, and the glycosaminoglycan moiety of the proteoglycans that form the epithelial cell glycocalyx. Upon the interaction of the vaginal isolate Lactobacillus salivarius Lv72 and HeLa cell cultures, the expression of oppA increased more than 50-fold over the following 30 min, with the overexpression enduring, albeit at a lower rate, for up to 24 h. Conversely, transcriptional analysis of 62 genes involved in proteoglycan biosynthesis revealed generalized repression of genes whose products catalyze different steps of the whole pathway. This led to decreases in the superficial concentration of heparan (60%) and chondroitin sulfate (40%), although the molecular masses of these glycosaminoglycans were higher than those of the control cultures. Despite this lowering in the concentration of the receptor, attachment of the Lactobacilli proceeded, and completely overlaid the underlying HeLa cell culture.

Glycocalyx regulates the strength and kinetics of cancer cell adhesion revealed by biophysical models based on high resolution label-free optical data

“…The basal zeta potential of HeLa cells was − 11.9 mV. ChrABC treatment significantly increased the zeta potential after 60 min treatment to − 10.2 mV, as a clear consequence of the removal of negatively charged glycocalyx component chondroitin sulfate, which is one of the dominant GAG on HeLa cells 54 . At 120 min no difference was seen anymore between treated cells and the 0 min control.…”

“…We employed the ChrABC enzyme to digest specific glycocalyx components during the adhesion event. ChrABC mainly digests the chondroitin sulfate components of the glycocalyx 21 , 54 , 65 . Lee et al digested a TE-1 cell line with 1 U/ml ChrABC.…”

“…We think that the main point in our work is the cleavage of the glycocalyx elements and how this affects cancer cell attachment and the validity of our observation is not less even if a decrease in HA also happens in addition to CS removal. Importantly, HeLa cell contains the polyanions heparan sulfate and CS 54 , but hyaluronic acid is not present on its surface 72 , 73 . Nonetheless, in future work, investigating hyaluronic acid of HeLa cells might prove important.…”

Antimicrobial properties of Lactobacillus cell‐free supernatants against multidrug‐resistant urogenital pathogens

“…The Lactobacillus adhesion to the HeLa cell layer was performed on microscope cover glasses and expressed as percentage adherence. Briefly, Lactobacillus cultures grown anaerobically for 48 h at 37°C in MRS broth (Oxoid) were harvested by centrifugation (5000 × g for 15 min, 4°C), and the cells were washed twice with a sterile solution of 0.85% NaCl (w/v) (Sigma‐Aldrich) diluted in RPMI 1640 medium at 5 × 108 CFU/ ml and incubated with a monolayer of HeLa for 1 h at 37°C (Martín et al, 2020 ; Mastromarino et al, 2002 ). After washes, the cells were fixed with 3 ml of methanol and stained with 3 ml of Giemsa stain solution (1:20; Carlo Erba, Milan, Italy) for 30 min at room temperature.…”

Characteristics of the Proteolytic Enzymes Produced by Lactic Acid Bacteria

“…They are coded by five genes ( oppD, oppF, oppB, oppC , and oppA ) with monocistronic property [ 74 ]. Opp consists of two OppD, OppF nucleotide binding molecules, two integral proteins of the OppB and OppC cell membrane, and an OppA protein that binds oligopeptide [ 75 , 76 ]. These are highly secured ATP-binding cassette transporters, from which obtained energy enables transport of casein-derived peptides across the cell membrane [ 72 , 77 ].This is an example of active transport, which requires energy input and involves specialized integral proteins coupling transport with the energy release process.…”