Marihuana is the most common street drug of abuse in the United States. Delta- 9-tetrahydrocannabinol (Delta-9-THC) is the major psychoactive component of marihuana and accounts for the majority of its immunosuppressive properties. The widespread use of marihuana has occurred at a time of the pandemic spread of sexually-transmitted viruses in the human population. The goal of this research is to define the effect of Delta-9-THC on T- lymphocyte functional competence to heroes simplex virus type 2 (HSV2). The B6C3F1 mouse will be employed for in vivo studies and as a source of splenocytes and T-lymphocytes. During the first phase of the study, the T-lymphocyte functional subset targeted by Delta-9-THC will be defined. A 3H-thymidine proliferative assay and a plaquing assay employing HSV2 antigen will be used to assess the effect of in vivo Delta-9-THC exposure on T-helper and T-suppressor cell activity. Reconstitution studies, using anti-L3T4 or anti-Lyt2 antibodies for selection for T-helper or T-suppressor cells, also, will be performed. Cytotoxic T-cell activity will be assessed by 51Cr-release from HSV2-infected mouse embryo fibroblast targets. In the second phase, the effect of the drug on early events of T-lymphocyte activation, as mediated by the inositol phosphate system, will be defined. In the third phase, the effect of the drug on T-lymphocyte: target cell adhesion will be determined using a LFA-1-dependent lymphocyte homotypic cluster assay, Nomarski microscopy, and scanning electron microscopy. In addition, immunochemical staining and transmission electron microscopy will be used to examine the effect of the drug on the lymphocyte cytoskeleton. The effect of the cannabinoid on macromolecular synthesis will be determined. A proliferative assay using IL-2-dependent CTLL-A2 cells will be used to assess interleukin 2 (IL-2) secretion. Monoclonal antibody to the IL-2 receptor will be used for immunofluorescence and flow cytofluorometry to define its cell-surface expression. The effect of Delta-9-THC on IFN-gamma will be assessed using a vesicular stomatitis virus plaque assay.
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