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Visualization of antigen-specific T cells has become an important tool in studying immune responses. The aim of this study was to analyze CMV-specific CD4+T cells in healthy and HIV-infected individuals. Peripheral blood mononuclear cells (PBMC) were examined for antigen-induced intracellular cytokine responses. We found significant numbers of CMV-specific CD4+T cells detectable in most CMV-IgG+HIV-1 infected individuals, whereas CMV-specific CD4+T cells could not be demonstrated in CMV-IgG-patients. Median frequency of CMV-specific CD4+T cells were lower in HIVinfected subjects who had been treated with highly active antiretroviral therapy (HAART) for more than 1 year than in untreated HIV-infected individuals. In patients under therapy for less than 1 year median CMV-specific CD4+T cell responder frequency was higher than in subjects treated for more than 1 year but lower than in untreated subjects. HIV suppression with HAART might lead to a progressive reduction of CMV-specific CD4+T cells indicating an efficient elimination of an opportunistic pathogen.