Cytotoxic T cell activation by Class I protein on cell-size artificial membranes: Antigen density and Lyt-2/3 function

The role of antigen surface density, and its relationship to the function of the Lyt-2/3 complex, in recognition and triggering of allospecific cytotoxic T lymphocyte (CTL) precursors has been studied by using a novel type of Class I protein-bearing artificial membrane. The cell-size membranes, termed pseudocytes (artificial cells), can be handled like cells but have a well-defined and easily quantitated surface composition. Class I antigen on these membranes stimulated generation of secondary in vitro allogeneic CTL responses as effectively as allogeneic spleen cells, provided that lymphokines were added to the cultures. Antigen density on the pseudocyte surfaces could be varied over a wide range and quantitated by papain cleavage and fluorescence-activated cell sorter analysis. Recognition and triggering of precursor CTL was found to be dramatically dependent on the surface density of antigen and displayed a marked threshold density requirement, below which little or no response occurred. Examination of the effects of anti-Lyt-2 antibody on responses to pseudocytes provided direct evidence for a reciprocal relationship between antigen density and susceptibility to antibody blockade. The results strongly suggest that antigen density is likely to have important biological consequences in control of immune responses. They also show that if Lyt-2/3 functions by interaction with a ligand, then that ligand is the Class I protein.