Cells from human immunodeficiency virus (HIV)-positive patients were evaluated for their in vitro responsiveness to recall antigen, alloantigen, and phytohemagglutinin (PHA) following the in vitro addition of interleukin (IL)-12 or anti-IL-10. Three-color flow cytometric analysis of CD4 and CD8 subsets was done to determine whether specific in vivo alterations in cell surface markers are associated with in vitro function changes. The results demonstrated a hierarchical response pattern to recall antigens versus alloantigen versus PHA, and these in vitro responses were associated with the number and activation status of CD4 cells. The in vitro addition of IL-12 or anti-IL-10 could restore antigen responses (HIV envelope peptides or influenza) in patients with 200-500 CD4 cells/microL; however, in patients with < 200 CD4 cells/microL, this improved response was limited to the influenza response. Studies of this nature may provide important insights into the role of cytokines in the natural history of HIV disease, and they suggest that immune therapy of this type may be most effective in patients who have more preserved immune systems.