title: Complement-activated vitronectin enhances the invasion of nonphagocytic cells by bacterial pathogens Burkholderia and Klebsiella.
abstract: Burkholderia pseudomallei is a serum-resistant Gram-negative bacterium capable of causing disseminated infections with metastatic complications. However, its interaction with nonphagocytic cells is poorly understood. We observed that exposure of B. pseudomallei and the closely related yet avirulent B. thailandensis to human plasma increased epithelial cell invasion by >20 fold. Enhanced invasion was primarily driven by a plasma factor, which required a functional complement cascade, but surprisingly, was downstream of C3 mediated opsonisation. Receptor blocking studies with RGD-domain containing peptide and αVβ3blocking antibody identified complement-activated vitronectin as the factor facilitating this invasion. Plasma treatment led to the recruitment of vitronectin onto the bacterial surface, and its conversion into the active conformation. Activation of vitronectin, as well as increased invasion, required the complement pathway and was not observed in C3 or C5 depleted serum. The integrin inhibitor cilengitide, currently in clinical trials as an anti-angiogenesis agent, suppresses plasma-mediated Burkholderia invasion by ~95%, along with a downstream reduction in intracellular bacterial replication. We extend these findings to serum-resistant Klebsiella pneumoniae as well. Thus, the potential use of commercially available integrin inhibitors as anti-infective agents during selective bacterial infections should be explored.
authors.affiliation: Department of Biochemistry, Yong Loo Lin School of Medicine, National University of Singapore, Singapore, 117597, Singapore.
authors.forename: Yi Han
authors.initials: YH
authors.lastname: Tan
authors.affiliation: Department of Biochemistry, Yong Loo Lin School of Medicine, National University of Singapore, Singapore, 117597, Singapore.
authors.forename: Akshamal M
authors.initials: AM
authors.lastname: Gamage
authors.affiliation: Department of Biochemistry, Yong Loo Lin School of Medicine, National University of Singapore, Singapore, 117597, Singapore.
authors.forename: Yunn-Hwen
authors.identifier: 0000-0001-9881-6881
authors.initials: YH
authors.lastname: Gan
mesh_terms: D016957:Burkholderia pseudomallei / Q000502:physiology*; D002460:Cell Line; D003165:Complement System Proteins / Q000378:metabolism*; D004705:Endocytosis; D004847:Epithelial Cells / Q000382:microbiology*; D054884:Host-Pathogen Interactions; D006801:Humans; D007155:Immunologic Factors / Q000378:metabolism*; D007711:Klebsiella pneumoniae / Q000502:physiology; D019096:Vitronectin / Q000378:metabolism*
chemical_list: D007155:Immunologic Factors; D019096:Vitronectin; D003165:Complement System Proteins