Date of Completion

1-31-2019

Embargo Period

1-29-2019

Keywords

Lyme, Borrelia, Macrophage, Innate, Immunity

Major Advisor

Juan Salazar

Associate Advisor

Justin Radolf

Associate Advisor

Melissa Caimano

Associate Advisor

Lynn Puddington

Field of Study

Biomedical Science

Degree

Doctor of Philosophy

Open Access

Open Access

Abstract

Lyme disease is a tick-borne illness caused by the spirochete Borrelia burgdorferi(Bb). It is believed that the robust inflammatory response induced by the host’s innate immune system is responsible for the clinical manifestations associated with Bb infection. The macrophage plays a central role in the immune response to many bacterial infections and is thought to play a central role in activation of the innate immune response to Bb. Previous studies have shown that following phagocytosis of spirochetes by macrophages, phagosome maturation results in degradation of Bband liberation of bacterial lipoproteins and nucleic acids, which are recognized by TLR2 and TLR8, respectively, and elicit MyD88-mediated phagosome signaling cascades. Bone marrow-derived macrophages (BMDMs) from MyD88-/-mice show significantly reduced spirochete uptake and inflammatory cytokine production when incubated with Bb ex vivo. Paradoxically, additional studies revealed that Bb-infected MyD88-/-mice exhibit inflammation injoint and heart tissues. To determine the contribution of MyD88 to macrophage-mediated spirochete clearance,we compared wildtype (WT) and MyD88-/-mice using a murine model of Lyme disease. MyD88-/-mice showed increased Bbburdens in hearts 28 days post infection, while H&E staining and immunohistochemistry showed significantly increased inflammation and greater macrophage infiltrate in the hearts of MyD88-/-mice. This suggests that Bb triggers MyD88-independent inflammatory pathways in macrophages to facilitate cell recruitment to tissues. Upon stimulation with Bbex vivo, WT and MyD88-/-BMDMs exhibit significant differences in bacteria uptake, suggesting that MyD88 signaling mediates cytoskeleton remodeling and the formation of membrane protrusions to enhance bacteria phagocytosis. A comprehensive transcriptome comparison in Bb-infected WT and MyD88-/-BMDMs identified a large cohort of MyD88-dependent genes that are differentially expressed in response toBb, including genes involved in actin and cytoskeleton organization (Daam1, Fmnl1).We also identified acohort of differentially-expressed MyD88-independent chemokines (Cxcl2, Ccl9) known to recruit macrophages. We identified master regulators and generated networks which model potential signaling pathways that mediate both phagocytosis and the inflammatory response. These data provide strong evidence that MyD88-dependent and -independent phagosomal signaling cascades in macrophages play significant roles in the ability of these cells to phagocytose Bb and mediate infection.

Download
COinS