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Immunopathology in Toxicology and Drug Development pp 343–369Cite as

Flow Cytometry and Immunophenotyping in Drug Development

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Part of the book series: Molecular and Integrative Toxicology ((MOLECUL))

Abstract

As increasing numbers of novel therapeutics impact immune responses either directly or indirectly, it is necessary to employ techniques to evaluate immune system alterations. Flow cytometry is an important tool that can be applied throughout drug development (e.g. early discovery to clinical monitoring) to evaluate immunobiological and immunotoxicological alterations resultant from therapies. Traditionally, flow cytometry has broad applications in assessing the identity of specific cell populations and their composition within bodily fluids and tissues. Application of cellular phenotype for routine immunophenotyping of cell populations has expanded to include enhanced immuno-phenotyping of cell subsets (e.g. Tregs, Th1, Th2, etc.) and functional immunophenotyping based on unique expression of cytokines, signaling markers and other markers identifying cell populations with specific functions. Although powerful in its ability to determine immunophenotyping, flow cytometry is increasingly being used to interrogate numerous other parameters including, cellular viability (e.g. apoptosis versus necrosis), cellular proliferation, phagocytosis, calcium influence, nucleic acid content, oxidative stress, signaling events and phosphorylation in signaling cascades and numerous others. Instrumentation, reagents and species-specific considerations, sample preparation and preservation, data acquisition, analysis and reporting and validation are all critical considerations in flow cytometry which are covered in this chapter. Ultimately, integration of flow cytometry with numerous other data sets (e.g. immunotoxicology, immunogenicity, pathology, etc.) are critical to develop a comprehensive picture of immune system alterations in drug development which have direction implications on human health.

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Abbreviations

APC:

Allophycocyanin

CD:

Clusters of differentiation; e.g. CD4, CD8

CFSE:

Carboxyfluorescein diacetate succinimidyl ester

CBC:

Complete blood count

DMSO:

Dimethylsulfoxide

FMO:

Fluorescence minus one

FACS:

Fluoroescent antibody cell sorting

FITC:

Fluoresceine isothiocyanate

GLP:

Good laboratory practice

IHC:

Immunohistochemistry

Ig:

Immunoglobulin; e.g. IgG, IgM

NTE:

Novel therapeutities

MHC:

Major histocompatibility complex

PBMCs:

Peripheral blood mononuclear cells

PE:

Phycoerythrin

PMT:

Photomultiplier tubes

Th1/Th2/Th17:

T helper 1, T helper 2 and T helper 17 cells

Treg:

Regulatory T cells

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  1. Charles River Laboratories, Inc., 1407 George Road, Ashland, OH, 44805, USA

    Tracey L. Papenfuss

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    George A. Parker

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Papenfuss, T.L. (2017). Flow Cytometry and Immunophenotyping in Drug Development. In: Parker, G. (eds) Immunopathology in Toxicology and Drug Development. Molecular and Integrative Toxicology. Humana Press, Cham. https://doi.org/10.1007/978-3-319-47377-2_6

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