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Detection
and isolation of circulating A2/Melan-A multimer+ CD8+ T cells
from PBMCs of healthy donor DR. (A) PBMCs from DR were stained
with the indicated A2/peptide multimersPE and anti-CD8
mAbFITC. (B) PBMCs were stimulated in vitro
with the Melan-A 26-35 peptide analog A27L (1 µM), cultured
for one week and stained as in A. A2/Melan-A multimer+ CD8+ T
cells present in this culture were isolated by cell sorting and
further expanded in PHA. (C) After an additional two weeks, the
resulting polyclonal Melan-A monospecific allorestricted population
was stained as in A. Numbers in quadrants represent the percentage
of multimer+ cells among CD8+ T cells.
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