What may be learned

The LDA approach is now widely used to estimate precursor frequencies for a wide range of immunological responses (T cell growth, cytokine secretion, helper T cells, pCTLs, B cells at different stages of ontogeny, etc.). It can be manipulated to define the absolute frequency as elicited by the strongest stimuli known, and the operational frequency, i.e. that actually used in the immune response. It can be used to study changes in functional precursors in tolerance (see Desaymard and Waldmann; Good and Nossal) and after immunization.

From the basic frequency data, one can design further, more revealing biological experiments dealing with cell-cell interactions. For example, the first evidence that T cells and B cells have to contact each other to collaborate arose when it was shown that a single T cell (from LDA) could only collaborate with one of many B cells offered to it. A modification of the procedure termed 'partition analysis' has allowed the investigation of T cells with inhibitory (suppressive) properties that are contained within a population of cells with helper properties.

In vivo, LDA can address a wide range of biological questions of precursor to product relationships. One particularly good example is the estimation of how many T stem cells are required to colonize a thymus lobe.

See also: Cytotoxic T lymphocytes; Cytotoxicity, assays for; Graft-versus-host reaction; Immune response in vitro-, Statistics, use in immunology.

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