PBMC-Derived T Cells

Daniel Lozano-Ojalvo, Rosina López-Fandiño, and Iván López-Expósito

Abstract T cell cultures are a valuable tool in food research to perform studies within the food allergy field. Their main applications aim to analyze immunological responses towards food protein antigens to gain further insights into the mechanisms responsible for the development of oral tolerance or for the triggering of food allergies. This chapter describes the main applications, isolation techniques, and culture conditions for PBMC-derived T cells. Furthermore, critical parameters of the model, together with the experimental read outs will be discussed.

Keywords T cells cultures • Food allergy • Immunomagnetic cell isolation • PBMCs • CD4+ T cells

Introduction and Origin

T lymphocytes or T cells are small (<10 μm) resting cells, which are generated in the bone marrow and migrate to the thymus where they become mature. Once matured, they enter the bloodstream and circulate to the secondary lymphoid organs, the sites of lymphocyte activation by the antigens. Actually, T cells are constantly recirculating between these organs until they encounter their specific antigen. Naïve T cells (T cells that have not yet met their specific antigen) bear antigen receptors specific for a single chemical structure, however lymphocytes in the body collectively carry millions of different receptor specificities. Only those that meet an antigen to which their receptors bind will be activated to proliferate and differentiate into effector cells (Janeway et al. 2005).

This chapter deals with T lymphocytes derived from peripheral blood mononuclear cells (PBMCs). T cells comprise, approximately, 45–70 % of PBMCs in human peripheral blood, with a count in healthy subjects in the range of 1 million cells/mL. For many years there was no function ascribed to these cells, until the 1960s, when it gradually became apparent that T lymphocytes were the key mediators of adaptive immunity. T cell cultures are a valuable tool in food research to perform studies within the food allergy field. Their main applications aim to analyze immunological responses towards food protein antigens to gain further insights into the mechanisms responsible for the development of oral tolerance or for the triggering of food allergies (Martino et al. 2012).

 
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