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Protocol for analyzing BCG-induced trained immunity in murine bone marrow-derived macrophages

STAR Protoc. 2024 Sep 20;5(3):103267. doi: 10.1016/j.xpro.2024.103267. Epub 2024 Aug 17.

Abstract

Bacillus Calmette-Guérin (BCG), the only licensed tuberculosis vaccine, provides non-specific protection against non-tuberculosis diseases that is mediated by trained immunity, a functional reprogramming mediated by innate immune memory. Here, we present a protocol for analyzing BCG-induced trained immunity in murine bone marrow-derived macrophages (BMDMs). We describe steps for preparing BCG single bacterial suspensions, isolating BMDM cells, and the training process. This protocol can assist researchers to conveniently utilize BMDM cells to study trained immunity. For complete details on the use and execution of this protocol, please refer to Xu et al.1.

Keywords: Cell culture; Cell isolation; Immunology; Microbiology; Model Organisms.

MeSH terms

  • Animals
  • BCG Vaccine* / immunology
  • Bone Marrow Cells / cytology
  • Bone Marrow Cells / immunology
  • Immunity, Innate / immunology
  • Immunologic Memory / immunology
  • Macrophages* / immunology
  • Mice
  • Mice, Inbred C57BL
  • Mycobacterium bovis / immunology
  • Trained Immunity

Substances

  • BCG Vaccine