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Allergy pp 17-31 | Cite as

T Cell: Primary Culture from Peripheral Blood

  • Monika RaulfEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 2020)

Abstract

Peripheral blood mononuclear cells (PBMCs) can be used to assess cell-mediated immunity in general or via antigen-specific stimulation to detect previous exposure to a variety of antigens/allergens and to monitor the response to immunotherapies. Peripheral blood is the most common source of mononuclear cells for in vitro cultures, although mononuclear cells can be obtained from other sources involved in the allergic reaction. PBMCs from individuals previously exposed to an antigen proliferate in vitro when stimulated with the specific antigen. Proliferation is measured by the incorporation of (3H)-thymidine into newly synthesized DNA. This parameter is often used as an endpoint of lymphocyte stimulation induced by antigen or antigen fragments (e.g., synthetic peptides), mitogens, or anti-CD3/anti-CD28 combinations. The aim of this chapter is to describe the culture of T cells obtained from peripheral blood and the collection of cell supernatants for cytokine measurement.

Key words

Peripheral blood mononuclear cells T-cell culture Proliferation Cytokines 

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Institute for Prevention and Occupational Medicine of the German Social Accident Insurance, Institute of the Ruhr-UniversityBochumGermany

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