Assessing Bacteria-Host Cell Interactions Using Biomimetic Beads

Take a multi-well plate containing a mammalian epithelial cell culture.

In the test wells, add a bacterial pathogen along with polymer beads coupled to bacterial surface proteins.

In the control wells, add the pathogen and beads lacking bacterial surface proteins. Incubate.

In the test wells, the beads mimic the bacterial surface and compete with the pathogen for binding to epithelial cell membrane phosphatidic acid, reducing bacterial attachment.

In the control wells, beads do not affect bacterial attachment.

Remove the media and wash with buffer to eliminate unbound components.

Incubate with a non-ionic detergent to lyse the cells, releasing membrane-attached bacteria and beads.

Pipette the lysate to create a homogeneous suspension, and serially dilute the homogenate using buffer.

Transfer the dilutions onto agar plates, incubate to allow colony formation, and count the colonies.

The test plate exhibits fewer colonies than the controls, indicating reduced bacterial-host interactions.

Prepare infection media by diluting bacterial cultures into colorless DMEM without additives.

Prewarmed to 37 degrees Celsius containing 10% volume to volume bead suspension. To give an MOI of 10, pr