Advertisement

Phagocytosis Assays for Borrelia burgdorferi

  • Juan Anguita
  • Ana Carreras-González
  • Nicolás Navasa
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1690)

Abstract

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 words

Phagocytosis Borrelia burgdorferi Bone marrow-derived macrophages Lentivirus Gene silencing 

Notes

Acknowledgments

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.

References

  1. 1.
    Shapiro ED (2008) Lyme disease. Adv Exp Med Biol 609:185–195CrossRefPubMedGoogle Scholar
  2. 2.
    Olson CM Jr, Bates TC, Izadi H, Radolf JD, Huber SA, Boyson JE, Anguita J (2009) Local production of IFN-γ by invariant NKT cells modulates acute Lyme carditis. J Immunol 182(6):3728–3734CrossRefPubMedPubMedCentralGoogle Scholar
  3. 3.
    Montgomery RR, Booth CJ, Wang X, Blaho VA, Malawista SE, Brown CR (2007) Recruitment of macrophages and polymorphonuclear leukocytes in Lyme carditis. Infect Immun 75(2):613–620CrossRefPubMedGoogle Scholar
  4. 4.
    Armstrong AL, Barthold SW, Persing DH, Beck DS (1992) Carditis in Lyme disease susceptible and resistant strains of laboratory mice infected with Borrelia burgdorferi. Am J Trop Med Hyg 47(2):249–258CrossRefPubMedGoogle Scholar
  5. 5.
    Barthold SW, Beck DS, Hansen GM, Terwilliger GA, Moody KD (1990) Lyme borreliosis in selected strains and ages of laboratory mice. J Infect Dis 162(1):133–138CrossRefPubMedGoogle Scholar
  6. 6.
    Naldini L, Blömer U, Gallay P, Ory D, Mulligan R, Gage FH, Verma IM, Trono D (1996) In vivo gene delivery and stable transduction of nondividing cells by a lentiviral vector. Science 272(5259):263–267CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media LLC 2018

Authors and Affiliations

  • Juan Anguita
    • 1
    • 2
  • Ana Carreras-González
    • 1
  • Nicolás Navasa
    • 1
  1. 1.CIC bioGUNE, Macrophage and Tick Vaccine LaboratoryParque Tenolágico de BizkaiaDerioSpain
  2. 2.Ikerbasque, Basque Foundation for ScienceBilbaoSpain

Personalised recommendations