Demonstration of HIV-encoded Proteins in Cultured and in Uncultured CD 4 Positive Mononuclear Cells from Hemophilia Patients Employing Monoclonal Antibodies against p 15, p 24, GP 41, GP 120, and Reverse Transcriptase
In the light of the large percentage of hemophilia patients with antibodies to HIV the identification of a specific virus infection in comparison to HIV antibody negative hemophilia patients has reached crucial importance. The low success rates of direct virus culture techniques together with the as yet low AIDS-di-sease rate observed in these patients separate these patients from the other main risk groups. Within this context, we studied the expression of CD3, CD4, CD8, and HLA class II antigens on fixed cells after PHA stimulation and Interleukin 2 propagation as well as on untreated blood mononuclear cells from healthy individuals and from hemophilia patients by fluorescence activated flow cytometry. Monoclonal antibodies thought to be specific for p 15, p 24, GP 41, GP 120, and for reverse transcriptase revealed a certain number of positive cells on all defined subpopulations analysed. From cell specimen of HIV antibody positive hemophilia patients cells with specific HIV antigens could be enriched by in vitro cultivation. Importantly the expression of virus-encoded antigens preceedes a cytopathic effect for several daVs. Current analyses aim at the prognostic relevance of low amounts of such viral HIV proteins selectively detectable by moAbs.directed to either p 24, GP 41, GP 120, and RT. The reliability, high sensitivity and monoclonal antibody dependent specificity of this newly developed method for the demonstration of HIV specific proteins make it applicable also for longitudinal surveys of hemophilia patients to assess a potential state of viremia or virus antigen processing in their mononuclear cells.