Phagocytosis Assays for Borrelia burgdorferi
Phagocytosis of Borrelia burgdorferi, the causative agent of Lyme disease, is a poorly understood process, despite its importance during the host immune response to infection. Thus, macrophages infiltrate the infected tissues, including the base of the heart and phagocytose the spirochete, therefore contributing to their elimination from infected tissues and leading to inflammation. An impaired bacterial clearance will result in bacterial persistence that may interfere with normal physiology of the heart, such as electrical signals from the heart, resulting in an impaired coordination of the beating of the heart or “heart block.” This chapter presents a protocol for establishing primary mouse macrophage cultures, a method for lentivirus silencing of primary cells, and a method for the in vitro study of macrophage phagocytosis of fluorescently labeled Borrelia burgdorferi.
Key wordsPhagocytosis Borrelia burgdorferi Bone marrow-derived macrophages Lentivirus Gene silencing
This work is supported by the grant SAF2015-65327-R from the Spanish Ministry of Economy and Competitiveness (MINECO). ACG is supported by a fellowship from the Basque Government. We thank the MINECO for the Severo Ochoa Excellence accreditation (SEV-16-0644).
We thank JD Radolf for providing the fluorescence-expressing B. burgdorferi.