Cytomegalovirus (CMV)-specific immunity was investigated in human immunodeficiency virus (HIV)-infected individuals. A case-control (1:2) study was performed with cases defined as having a history of CMV end-organ disease (n=15) and controls (n=30) matched by current CD4(+) T cell count. CMV-specific CD8(+) T cells responses were quantified using the high throughput Quantiferon-CMV test (Cellestis, Melbourne, Australia). 40/44 (91%) had a positive Quantiferon-CMV test and the magnitude of response to CMV peptides correlated significantly with response to mitogen (p<0.0001) but not with CD4(+) T cell count at the time of testing, CD4(+) T cell nadir or HIV viral load. Cases had a significantly lower Quantiferon-CMV test than controls but there was no significant difference in response to mitogen or other antigens. In individuals with a history of CMV disease, CMV-specific CD8(+) T cell responses are reduced even in the setting of CD4(+) T cell reconstitution.