Production of TH1 and TH2 cell lines and clones.

This unit describes protocols for the generation of polyclonal T(H)1 and T(H)2 cell lines from naive CD4(+) T cells as well as for generation of antigen-specific cell lines from TCR-transgenic mice and antigen-specific T cell clones from primed mice. Also described are methods for the preparation and maintenance of alloreactive murine helper T (T(H)) lymphocyte and cytotoxic T lymphocyte (CTL) clones using the limiting dilution technique, as well as derivation of T(H) clones reactive with soluble protein antigens, including a method for the selection of either T(H)1 or T(H)2 lymphocyte subsets. These two subsets of T(H) cells exhibit helper function in different ways and can be distinguished by the patterns of cytokines they synthesize. Support protocols describe a micromanipulation method for cloning T cells and a roadmap for using protocols published elsewhere in this series to assess cytokine production by T cell clones and lines.