Clonality of PM1#11 crossreacting response to GAD65 and hCMV. T cells
were prepulsed overnight with either GAD65 339–352 or hCMV major
DNA-binding protein 674–687 (10 μM). After washing the T cells,
cells were stimulated with DR3-matched irradiated peripheral blood
mononuclear cells and either GAD65 339–352 or hCMV major DNA-binding
protein 675–687 (1 μM) and cultured as described. As a control for
the ability to proliferate, prepulsed cells were stimulated with IL-2.
In a negative control experiment (data not shown), PM1#11 was mixed
with a coxsackie B4 virus-specific T cell clone and prepulsed with
either the coxsackie B4 virus-derived epitope or GAD65 339–352. The
response of the T cell mixture, after washing, to GAD65 339–352 was
completely abrogated after prepulsing with GAD65 339–352, but not
after prepulsing with coxsackie B4A virus-derived epitope. In all
experiments, responses are the means ± SD of triplicate tests.
Mean background of triplicates after prepulsing never exceeded 500
cpm.