Intravital Imaging of Vascular Transmigration by the Lyme Spirochete: Requirement for the Integrin Binding Residues of the B. burgdorferi P66 Protein

• DOI: 10.1371/journal.ppat.1005333
• Publication Date: 2015-12-18T18:43:44Z
• Authors: Devender Kumar; Laura C. Ristow; Meiqing Shi; Priyanka Mukherjee; Jennifer A. Caine; Woo-Yong Lee; Paul Kubes; Jenifer Coburn; George Chaconas
• Keywords: Mice, Knockout; 0301 basic medicine; Lyme Disease; Mice, Inbred BALB C; Microscopy, Confocal; Intravital Microscopy; QH301-705.5; Transendothelial and Transepithelial Migration; Porins; RC581-607; Disease Models, Animal; Mice; 03 medical and health sciences; Bacterial Proteins; Borrelia burgdorferi; Host-Pathogen Interactions; Animals; Immunologic diseases. Allergy; Biology (General); Research Article

Vascular extravasation, a key step in systemic infection by hematogenous microbial pathogens, is poorly understood, but has been postulated to encompass features similar to vascular transmigration by leukocytes. The Lyme disease spirochete can cause a variety of clinical manifestations, including arthritis, upon hematogenous dissemination. This pathogen encodes numerous surface adhesive proteins (adhesins) that may promote extravasation, but none have yet been implicated in this process. In this work we report the novel use of intravital microscopy of the peripheral knee vasculature to study transmigration of the Lyme spirochete in living Cd1d-/-mice. In the absence of iNKT cells, major immune modulators in the mouse joint, spirochetes that have extravasated into joint-proximal tissue remain in the local milieu and can be enumerated accurately. We show that BBK32, a fibronectin and glycosaminoglycan adhesin of B. burgdorferi involved in early steps of endothelial adhesion, is not required for extravasation from the peripheral knee vasculature. In contrast, almost no transmigration occurs in the absence of P66, an outer membrane protein that has porin and integrin adhesin functions. Importantly, P66 mutants specifically defective in integrin binding were incapable of promoting extravasation. P66 itself does not promote detectable microvascular interactions, suggesting that vascular adhesion of B. burgdorferi mediated by other adhesins, sets the stage for P66-integrin interactions leading to transmigration. Although integrin-binding proteins with diverse functions are encoded by a variety of bacterial pathogens, P66 is the first to have a documented and direct role in vascular transmigration. The emerging picture of vascular escape by the Lyme spirochete shows similarities, but distinct differences from leukocyte transmigration.