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RESEARCH PRODUCT

2 Isolation of T Cells and Establishment of T-cell Lines and Clones

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Publisher Summary This chapter elaborates the isolation of T cells and the establishment of T-cell lines and clones. The study of human T cells is best performed using purified cells because the presence of other cell types may have indirect effects on T-cell function. For any kind of functional assay on T-cell specificity, antigen-presenting cells are necessary. T cells appear to play a major role in the development, maintenance, and also resolution of these forms of bacteria-associated arthritides. The E-rosetting technique describes a procedure for separating T cells and non-T cells from a population of MNCs. This method is based on the ability of human T cells to bind to sheep erthrocytes through their CD2 molecule. T cells expressing particular cell surface markers, such as the CD4, CD8, αβ- TCR or γδTCR molecules, can be selected by their capacity to bind to antibody-coated plastic plates. T-cell lines are the cultures of T lymphocytes grown by repeated cycles of stimulation, usually with antigen, antigen-presenting cells, and growth factors, such as interleukin-2. A T-cell clone is a continuously growing line of T cells derived from a single progenitor cell. The generation of T-cell clones helps in the identification of even minor fractions of T-cell populations and the investigation of effector functions that are stable within each line.