Insights into In Vitro Immunological Cytotoxicity Studies

Abstract

Cell‐mediated cytotoxicity is a critical response in maintaining and regulating good immune health by the highly selective removal of either virally infected or cancerous cells. There are multiple mechanisms that can be deployed by an effector cell with cytotoxic capabilities that are dependent on a number of factors such as immune context and target cell type. Insights into how, why and when an effector cell utilises a specific mode of action for target cell removal can be gained using a variety appropriate in vitro assays. In turn, data gathered from such in vitro systems can support recapitulation of physiological responses for development of novel therapeutic options.

Key Concepts

  • T and NK cells mediated cytotoxicity is required for immune regulation and clearance of pathogen‐infected cells.
  • Perforin, granzymes and death receptors are essential for cell‐mediated cytotoxicity.
  • Development of accurate in vitro assays to study cytotoxicity can help further understand physiological immune responses.
  • A better in vitro recapitulation of ADCC can be used to improve vaccines and vaccine responses.
  • Cytotoxicity in vitro assays can also be used in a clinical setting to diagnose underlying Immunodeficiencies.

Keywords: cytotoxicity; antibody‐dependent cellular cytotoxicity; granzymes; perforin; caspase; chromium release; death receptor; degranulation; HLH; ALPS

Figure 1. Schematic of perforin‐/granzyme‐mediated cytotoxicity. Granules are exocytosed from the effector cell(s) and internalised into the target cells via perforin‐mediated pores. The contents from the granules is then released and depending on the granzyme type distinct signalling cascades are activated to induce cellular apoptosis.
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Further Reading

Adan A, Kiraz Y and Baran Y (2016) Cell proliferation and cytotoxicity assays. Current Pharmaceutical Biotechnology 17 (14): 1213–1221.

Fassy J, Tsalkitzi K, Goncalves‐Maia M, et al. (2017) A real‐time cytotoxicity assay as an alternative to the standard chromium‐51 release assay for measurement of human NK and T cell cytotoxic activity. Current Protocols in Immunology 118: 7.42.1–7.42.12.

Iwahori K, Shintani Y, Funaki S, et al. (2019) Peripheral T cell cytotoxicity predicts T cell function in the tumor microenvironment. Scientific Reports 9 (1): 2636.

Tahara H, Matsuda S, Yamamoto Y, et al. (2017) High‐content image analysis (HCIA) assay has the highest correlation with direct counting cell suspension compared to the ATP, WST‐8 and Alamar blue assays for measurement of cytotoxicity. Journal of Pharmacological and Toxicological Methods 88 (Pt 1): 92–99.

Wang W, Erbe AK, Hank JA, et al. (2015) NK cell‐mediated antibody‐dependent cellular cytotoxicity in cancer immunotherapy. Frontiers in Immunology 6: 368.

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Panchal, Neelam, Gilmour, Kimberly, and Booth, Claire(Jul 2019) Insights into In Vitro Immunological Cytotoxicity Studies. In: eLS. John Wiley & Sons Ltd, Chichester. http://www.els.net [doi: 10.1002/9780470015902.a0001178.pub2]