Isolation and development of murine T-lymphocyte hybridomas and clonal cell lines

Publisher Summary This chapter describes the isolation and development of murine T-lymphocyte hybridomas and clonal cell lines. To study T lymphocytes that mediate many important functions of the immune system, the analysis of clonal cell populations has been necessary because T cells and B cells express clonally distributed antigen receptors, reflecting their clonal specificities for antigens. Cloning does not only represent an approach to studying individual T cells, but for studying collections of individual T-cell clones as well. The study of the collections of T-cell clones has been used to understand mixed cell populations, T-cell receptor (TCR) repertoires, and the development of immune responses. Cloned T cells readily permit the establishment of correlations among separate properties, such as cytokine secretion and the expression of cell surface markers or the pairing of heterodimeric chains of TCRs. The chapter introduces methods for isolating T cells from different tissues and describes the production of T-cell lines, T-cell clones, and T-cell hybridomas and transfectomas. T-cell proliferation assays, based on flow cytometry, are also described in the chapter.