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* Vaccine Research Center and
National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20892;
California National Primate Research Center, Center for Comparative Medicine and Division of Infectious Diseases, and
Department of Internal Medicine, University of California School of Medicine, Davis, CA 95616
In recent years, the quantification of T cell responses to pathogens or immunogens has become a common tool in the evaluation of disease pathogenesis or vaccine immunogenicity. Such measurements are usually limited to enumerating IFN-
-producing cells after ex vivo stimulation with Ag, but little is known about the phenotype or complete functional repertoire of the Ag-specific cells. We used 12-color flow cytometry to characterize Ag-specific T cells elicited by vaccines or natural infection to determine lineage and differentiation status as well as the capacity to produce four cytokines (IFN-, TNF-
, IL-2, and IL-4) and a chemokine (MIP1
). As expected, responding cells had a typical memory phenotype; however, the cytokine profiles associated with the responses were highly complex. The pattern of cytokine coexpression in response to specific Ags was a skewed subset of the complete repertoire (revealed by polyclonal stimulation). We found significant differences in the patterns of cytokines elicited by vaccination (where IFN- was by far a subdominant response) vs natural infection; in addition, there was fairly significant intersubject variation. Our findings illustrate the limitation of the evaluation of immune responses using single functional measurements (such as IFN-); in fact, it is likely that sensitive evaluation of Ag-specific T cells will require the coordinate measurement of several cytokines. The presence and variability of these complex response profiles introduce the possibility that selective functional expression patterns may provide correlates for vaccine efficacy or disease progression.
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