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Methods Mol Biol. 2017;1591:101-119. doi: 10.1007/978-1-4939-6931-9_8.

Endocrine Regulation of Lymphocyte Trafficking In Vitro.

Author information

1
Institute of Cardiovascular Sciences, College of Medical and Dental Sciences, University of Birmingham, Vincent Drive, Edgbaston, Birmingham, West Midlands, B15 2TT, UK.
2
Institute of Cardiovascular Sciences, College of Medical and Dental Sciences, University of Birmingham, Birmingham, B15 2TT, UK.
3
Institute of Inflammation and Ageing, College of Medical and Dental Sciences, University of Birmingham, Birmingham, B15 2TT, UK. h.m.mcgettrick@bham.ac.uk.

Abstract

Lymphocyte recruitment in inflammation can be influenced by many molecules including cytokines, chemokines, and adipokines. In our lab, we have examined the effects of the adipokines leptin and adiponectin on lymphocyte migration, and observed modulation of this process. Lymphocyte behavior can be assessed in the lab under static conditions, or can be studied under flow, simulating in vivo conditions. In this chapter, in vitro methods for analyzing adhesion and migration of lymphocytes isolated from blood are described in detail. In static adhesion and migration assays, lymphocytes are allowed to settle on top of endothelial cell monolayers cultured in plates for a desired period of time. In the flow-based assay, lymphocytes are perfused over the endothelium at a continuous rate through microchannels which are commercially available. Depending on the choice of method employed, the efficiency of lymphocytes to adhere to and migrate across the endothelial cell monolayer under different conditions can be evaluated. Static assays are less complex and are of higher throughput. However, these assays provide less detailed information regarding lymphocyte behaviors. On the other hand, the flow-based assays are more difficult to perform, but are more physiologically relevant due to the presence of flow and yield more detailed information about lymphocyte activities such as capture, immobilization, and migration in real-time.

KEYWORDS:

Adhesion; Adipokines; Adiponectin; Cell culture; Cytokines; Endothelial cells; Flow; Inflammation; Leptin; Lymphocyte; Migration

PMID:
28349478
DOI:
10.1007/978-1-4939-6931-9_8
[Indexed for MEDLINE]

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