Effective reconstitution of cellular immunity following hematopoietic stem cell transplantation (HCT) is thought to be important for protection from the morbidity caused by cytomegalovirus (CMV) reactivation and disease. This review critically discusses current methods for assessment of CMV-specific cellular immune responses, with emphasis on flow cytometry-based methodologies such as MHC-I and MHC-II tetramer staining and intracellular cytokine assays. The advantages and weaknesses of these assays are considered in comparison to traditional immunologic techniques. Application of these newer methodologies has provided insight into the dynamics of the levels of CMV-specific CD4(+) and CD8(+) T-lymphocytes following HCT, and into the sources and diversity of these cells. Data from preliminary clinical studies suggest that CMV-specific CD8(+) T-lymphocyte levels greater than 1 x 10(7)/L of peripheral blood may correlate with protection from CMV disease. Studies on the functional phenotypes of CMV-specific CD8(+) T-lymphocytes such as cytokine production, degranulation, and cytotoxicity have indicated that these cells are heterogeneous with regard to these properties. Future research will focus on establishing whether any of these immunologic assays will serve as a correlate of protection and inform as to which patients are at high risk for CMV reactivation and disease. Identification of an informative assay may allow its incorporation into standard clinical practice for monitoring HCT patients.