Clonal Culture Models of T-cell Senescence

Abstract

Studying human T-cell senescence is mostly limited to investigations on peripheral blood ex vivo or on cultured cells in vitro. In both cases, single cell analysis is challenging and many age-associated alterations described are the result of changes in the proportions of the ever-increasing numbers of different T-cell subsets rather than changes to the cells per se. One model avoiding this problem utilises monoclonal populations cultured long-term in vitro. Such T-cell clones (TCC) can be maintained without oncogenic transformation by intermittent antigen restimulation in the presence of growth factors. However, TCC possess finite lifespans (which vary greatly from clone to clone). This TCC model can be used to investigate many aspects of the processes of clonal expansion and contraction essential for adaptive immunity, including biomarker discovery at the genomic, proteomic and functional levels, and to test interventions of possible clinical utility. This chapter describes techniques for the production and maintenance of human TCC in vitro, the impact of culture conditions and oxygen levels on lifespan, and the application of genomic and proteomic analyses in this model.