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Quantitative Phenotypic Analysis of Drug Sequestering Macrophage Subpopulations

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Quantitative Analysis of Cellular Drug Transport, Disposition, and Delivery

Part of the book series: Methods in Pharmacology and Toxicology ((MIPT))

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Abstract

Macrophages reside in every tissue of the body and are the first-line of defense of the immune system. Their function is to entrap apoptotic cells, pathogens, and other particles and produce immune effector cytokines. It has also been shown that macrophages sequester xenobiotics, particularly lysosomotropic agents. One of the well-known and best studied lysosomotropic agents is the weakly basic antibiotic clofazimine. Interestingly, not all macrophages within a population, such as those present in a particular tissue or organ, accumulate xenobiotics to the same extent. Within every general population of macrophages, there is a distinct subpopulation of macrophages that express the xenobiotic sequestering phenotype. Thus, in this chapter, we explain how one can utilize clofazimine as a probe to explore the physical and biological markers that can ultimately distinguish this xenobiotic sequestering subpopulation of macrophages, specifically among the alveolar macrophages isolated from the lungs of mice treated with oral clofazimine for a period of several weeks. The experimental approach and the rationale behind choosing each marker is also explained. As a result, out of various biological markers (i.e., TLR2, TLR4, CLC7, TFEB, p65, V-ATPase) and physical markers (i.e., cell area and size), the physical markers were most closely associated with the xenobiotic sequestering subpopulation of macrophages. Other investigated molecular markers showed no statistically significant association with the xenobiotic sequestering phenotype. Some markers exhibited differences, but the variability in the phenotype rendered them a less reliable marker as compared to the associated increase in cell area and size.

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Authors and Affiliations

  1. Department of Pharmaceutical Sciences, University of Michigan, College of Pharmacy, Ann Arbor, MI, USA

    Mikhail D. Murashov

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  1. Mikhail D. Murashov
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Correspondence to Mikhail D. Murashov .

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Editors and Affiliations

  1. College of Pharmacy, University of Michigan, Ann Arbor, MI, USA

    Gus R. Rosania

  2. Department of Chemical Engineering, University of Michigan, Ann Arbor, MI, USA

    Greg M. Thurber

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Murashov, M.D. (2021). Quantitative Phenotypic Analysis of Drug Sequestering Macrophage Subpopulations. In: Rosania, G.R., Thurber, G.M. (eds) Quantitative Analysis of Cellular Drug Transport, Disposition, and Delivery. Methods in Pharmacology and Toxicology. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1250-7_7

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  • DOI: https://doi.org/10.1007/978-1-0716-1250-7_7

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-1249-1

  • Online ISBN: 978-1-0716-1250-7

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