T Cell: Primary Culture from Peripheral Blood

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.