In the pediatric human immunodeficiency virus type-1 (HIV-1) infection, the presence of cytotoxic T lymphocytes (CTL) is associated with a slow progression to AIDS. The secretion of cytokines by CTLs may be critical in the control of viral infection. We used the combination of cell surface and intracellular staining to study the functionality of tetramer binding CD8+ T cells recognizing two HIV-1 immunodominant epitopes, in peripheral blood mononuclear cells from HIV-1-infected children. A fraction of tetramer positive CD8+ T cells produce cytokines (IFN-gamma, TNF-alpha) or chemokines (CCL4, CCL5) after ex vivo stimulation with the cognate peptide. There was a negative correlation between the plasma viral load and the percentage of CD8+ Tetramer Gag+ T cells secreting IFN-gamma. This is the first report in the context of pediatric HIV-1 infection showing that only a fraction of HIV-1-specific CD8+ T cells have the capacity to produce cytokines and chemokines implicated in their antiviral functions.