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By
From the Molecular Immunology Group, Institute of Molecular Medicine, Nuffield Department of
Clinical Medicine, University of Oxford, John Radcliffe Hospital, Oxford OX3 9DU, United
Kingdom
The nature of the CD8+ T cells that underlie antiviral protective immunological memory in
vivo is unclear. We have characterized peptide-specific CD8+ T lymphocytes directly ex vivo
from peripheral blood in humans with past exposure to influenza virus, using single cell interferon 
(IFN-) release as a measure of effector function. In individuals in the memory state
with respect to influenza virus infection, unrestimulated antigen-specific CD8+ T cells displayed IFN- release within 6 h of antigen contact, identifying a population of memory CD8+
T cells that exhibit effector function without needing to divide and differentiate over several
days. We have quantified circulating CD8+ effector T cells specific for six different MHC class
I-restricted influenza virus epitopes. Enumeration of these CD8+ T cells gives frequencies of
peptide-specific T cells that correlate with, but are in general severalfold higher than, CTL precursor frequencies derived from limiting dilution analysis, indicating that this novel population
of memory CD8+ T cells has hitherto been undetected by standard means. The phenotype of
these cells, which persist at a low frequency long after recovery from an acute viral infection,
suggests that they play a role in protective immunological memory.
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