Acquired Immunodeficiency Syndrome (AIDS) is characterized by reduced numbers and functional impairment of T lymphocytes. The major emphasis of this investigation will be an examination of the in vitro expandability of peripheral blood lymphocytes from patients with AIDS using protocols designed to examine specific molecular events (e.g. phospholipid and calcium metabolism and kinase activation). These events, which are crucial to Tcell proliferation and functional development, will be investigated using probes of the cell surface (e.g. phospholipase C) and cytosol (e.g. calcium ionophores and protein kinase C modulators). The populations studied will include individuals with AIDS, healthy individuals who are HIV antibody positive, and healthy controls. The lymphocytes from these subjects will be characterized for functionally important structures (e.g. T4, T8, T3, IL-2 receptor) before and after the T-cell expandability studies. The various T cell populations will also be assessed for in vitro cytotoxic function against Epstein Barr Virus transformed autologous B cells. Analysis of retrovirus integration and replication and the use of specific viral inhibitors will concomitantly assess the effect of such agents on Tcell proliferation and HIV replication. The in vitro addition of selected opioids (agonists and antagonists) and cannabinoid compounds will be used to further examine the impact of drugs of abuse in this paradigm. The design and analysis of these protocols will be accomplished via the use of response surface methodology (RSM). The use of RSM will allow for developing optimum expandability protocols and the mechanistic exploration of those factors (e.g. HIV, drugs of abuse) which impact on AIDS related Tcell functionality. The understanding of those factors which result in Tcell dysfunction and the ability to reverse these deficits in vitro could result in the eventual development of adoptive immunotherapy procedures for the treatment of AIDS.
