Human T Cell Clones in Long-Term Culture as Models for the Impact of Chronic Antigenic Stress in Aging

This chapter discusses techniques for the production and maintenance of human T cell clones, their growth characteristics and longevity in vitro, culture age-associated changes of parameters such as telomere lengths, DNA damage, and so on. Cloning peripheral T cells from young healthy donors can be performed with a high cloning efficiency (CE), suggesting that these particular cloning conditions are favorable for outgrowth of a majority of T cells. The CE was equally high when the cells were derived from a patient with chronic myelogenous leukemia, a donor putatively subjected to a high level of antigenic stress. CE was also similar in the extra-thymic T cell differentiation cultures employed. According to the protocol for cloning, for ease of handling and as a precaution against contamination, stack plates and aluminum foil should be used. Incubate for about a week and then examine the plates using an inverted microscope. T cell functions are mediated by intracellular signaling via surface receptors such as the antigen receptor and the costimulator CD28 from the membrane through the cytoplasm to the nucleus.