Cell Adhesion Assays to Investigate Leptospiral Protein Interactions with ECM, Cell Receptors, and Cultured Mammalian Cells: Methods for Understanding Host-Leptospira Interactions.

Pathogenic bacteria must overcome host barriers and survive to establish infection. Adhesion to host cells and extracellular matrix (ECM) components is a critical step in this process, as it facilitates colonization and invasion. Pathogenic Leptospira species have been shown to bind both to host cell receptors and to ECM molecules, contributing to infection. Previous studies have mainly reported in vitro interactions of recombinant Leptospira interrogans proteins with ECM components. Our work extends these studies by focusing on two major classes of macromolecules that constitute the mammalian ECM: glycosaminoglycans (GAGs) and fibrous proteins such as collagens, elastin, fibronectin, and laminin. Additionally, we have investigated interactions with various integrin cell receptors and examined the binding of recombinant proteins to cultured mammalian cell. In this chapter, we describe the established protocols to investigate the interactions between putative leptospiral adhesins and host components and mammalian cells.