Polyclonal and Antigen-specific Responses of T Cells and T Cell Subsets

Evaluation of the functional responses of T cells is of importance in determining the mechanism(s) of immunodeficiency resulting from chronic alcohol abuse and other conditions that lead to immune dysfunction. Mice that are chronically exposed to 20% (w/v) ethanol in water develop immunodeficiency and have T cells with abnormal activation profiles, reduced total numbers, increased CD4/CD8 ratios, and an increased memory/na�ve phenotype ratio. These cells also have abnormal antigen-specific responses after inoculation of the ethanol mice with model infectious organisms. Study of the functional abnormalities of these cells requires a reliable system that can present appropriate activation stimuli in vitro for the generation of polyclonal or antigen-specific responses in enriched or purified T cells, free of the influence of previously ethanol exposed accessory cells. In this chapter, we describe protocols to assess the T cell response to polyclonal stimulation through the T cell receptor and the use of a model infectious disease bacterium, Listeria monocytogenes , that allows evaluation of the T-cell response to specific peptide epitopes of the bacterium after previous inoculation.