Chapter 6
Macrophage Infection Models for Mycobacterium
tuberculosis
Benjamin K. Johnson, Sean M. Thomas, Andrew J. Olive,
and Robert B. Abramovitch
Abstract
Mycobacterium tuberculosis colonizes, survives, and grows inside macrophages. In vitro macrophage infection models, using both primary macrophages and cell lines, enable the characterization of the pathogen
response to macrophage immune pressure and intracellular environmental cues. We describe methods to
propagate and infect primary murine bone marrow-derived macrophages, HoxB8 conditionally immortalized myeloid cells, Max Planck Institute alveolar macrophage-like cells, and J774 and THP-1 macrophagelike cell lines. We also present methods on the characterization of M. tuberculosis intracellular survival and
the preparation of infected macrophages for imaging.
Key words Mycobacterium tuberculosis, Primary macrophages, Macrophage-like cell lines, Intracellular infection methods
1 Introduction
A hallmark of Mycobacterium tuberculosis pathogenesis is the ability
to colonize, survive, and replicate within macrophages [1]. Understanding the mechanisms of M. tuberculosis–macrophage interactions promises to identify physiological pathways that are central to
both bacterial virulence and host immunity. The macrophage niche
represents a complex and dynamic environment that cannot be
easily replicated by studying extracellular M. tuberculosis in bacterial
growth media; therefore, several ex vivo M. tuberculosis–macrophage infection models have been established. Primary murine
bone marrow-derived macrophages (BMDMs) have proven to be
a useful model system because they can be easily cultured and
expanded to large numbers of cells. Primary macrophages isolated
from knockout mice enable the study of M. tuberculosis in macrophages lacking specific features of host immunity or to characterize
Tanya Parish and Anuradha Kumar (eds.), Mycobacteria Protocols, Methods in Molecular Biology, vol. 2314,
https://doi.org/10.1007/978-1-0716-1460-0_6, © Springer Science+Business Media, LLC, part of Springer Nature 2021
167
Macrophage Infection Models for Mycobacterium
tuberculosis
Benjamin K. Johnson, Sean M. Thomas, Andrew J. Olive,
and Robert B. Abramovitch
Abstract
Mycobacterium tuberculosis colonizes, survives, and grows inside macrophages. In vitro macrophage infection models, using both primary macrophages and cell lines, enable the characterization of the pathogen
response to macrophage immune pressure and intracellular environmental cues. We describe methods to
propagate and infect primary murine bone marrow-derived macrophages, HoxB8 conditionally immortalized myeloid cells, Max Planck Institute alveolar macrophage-like cells, and J774 and THP-1 macrophagelike cell lines. We also present methods on the characterization of M. tuberculosis intracellular survival and
the preparation of infected macrophages for imaging.
Key words Mycobacterium tuberculosis, Primary macrophages, Macrophage-like cell lines, Intracellular infection methods
1 Introduction
A hallmark of Mycobacterium tuberculosis pathogenesis is the ability
to colonize, survive, and replicate within macrophages [1]. Understanding the mechanisms of M. tuberculosis–macrophage interactions promises to identify physiological pathways that are central to
both bacterial virulence and host immunity. The macrophage niche
represents a complex and dynamic environment that cannot be
easily replicated by studying extracellular M. tuberculosis in bacterial
growth media; therefore, several ex vivo M. tuberculosis–macrophage infection models have been established. Primary murine
bone marrow-derived macrophages (BMDMs) have proven to be
a useful model system because they can be easily cultured and
expanded to large numbers of cells. Primary macrophages isolated
from knockout mice enable the study of M. tuberculosis in macrophages lacking specific features of host immunity or to characterize
Tanya Parish and Anuradha Kumar (eds.), Mycobacteria Protocols, Methods in Molecular Biology, vol. 2314,
https://doi.org/10.1007/978-1-0716-1460-0_6, © Springer Science+Business Media, LLC, part of Springer Nature 2021
167