A group of 18 individuals is described that manifest persistent (follow up between 4 and 57 months) expansions (ranging from 10 to 70%) of CD4+ CD8+ peripheral T cells. In nine individuals there was no apparent underlying disease whereas the remaining displayed a wide range of unrelated diseases. Analysis of the cell surface immunophenotype showed that all individuals express high densities of CD2, CD3, CD5, and TCR alpha/beta on their CD4+ CD8+ subset. On the contrary, these cells did not express CD25, CD38, CD71, HLA-DR, and the beta chain of the interleukin-2 receptor (p75) but expressed high density of CD29 and low density or absence of CD7. CD8 was "dim" in most individuals and consisted only of the CD8 alpha chain isoform. These individuals could be further divided into two subgroups based on the following parameters: in eleven cases CD4+ CD8+ cells had an LGL morphology and expressed the CD11b, CD56 and CD57 NK-associated antigens; in addition, a high percentage of CD4+ CD8- cells from the same individuals also expressed NK-associated antigens on their cell surface (subgroup A). The CD4+ CD8+ cells of the remaining individuals had small lymphocyte morphology, expressed little or no CD11b and CD56 and their CD4+ CD8- cells did not express NK-associated antigens at all (subgroup B). Finally, CD8 was "bright" on the CD4+ CD8+ cells of three individuals and in these cells also the CD8 beta chain isoform was present (subgroup C). Southern blot hybridization of genomic DNA with a T-cell receptor beta probe revealed that of the sixteen individuals examined four had a monoclonal, seven an oligoclonal, and five a polyclonal molecular configuration. The present data indicate that persistent CD4+ CD8+ expansions represent a novel heterogeneous subset of T cells.