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Department of Immunology, St. Jude Childrens Research Hospital, Memphis, TN 38105
Screening with the flow cytometric IFN-
assay has led to the
identification of a new immunogenic peptide (SSYRRVPGI) from the
influenza PB1 polymerase (PB1703711) and a mimotope
(ISPLMVAYM) from the PB2 polymerase (PB2198206).
CD8+ T cells specific for KbPB1703
make both IFN-
and TNF-
following stimulation with both peptides.
The CD8+ KbPB1703+
population kills PB2198-pulsed targets, but cell lines
stimulated with PB2198 neither bind the
KbPB1703 tetramer nor become CTL. This
CD8+KbPB1703+
population is prominent in the primary response to an H3N2 virus,
although it is much less obvious following secondary challenge of
H1N1-primed mice. Even so, we can now account for >40% of the
CD8+ T cells in a primary influenza pneumonia and >85% of
those present after H3N2
H1N1 challenge. Profiles of IFN-
and
TNF-
staining following in vitro stimulation have been traced for
the four most prominent influenza peptides through primary and
secondary responses into long-term memory. The
DbNP366 epitope that is immunodominant after
the H3N2
H1N1 challenge shows the lowest frequencies of
CD8+ IFN-
+TNF-
+ cells for >6
wk, and the intensity of IFN-
staining is also low for the first 3
wk. By 11 wk, however, the IFN-
/TNF-
profiles look to be similar
for all four epitopes. At least by the criterion of cytokine
production, there is considerable epitope-related functional diversity
in the influenza virus-specific CD8+ T cell response. The
results for the KbPB1703 epitope and the
PB2198 mimotope also provide a cautionary tale for those
using the cytokine staining approach to identity antigenic
peptides.
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