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Program in Molecular Cardiobiology, Boyer Center for Molecular Medicine, Yale University School of Medicine, New Haven, CT 06510
We have shown previously that cultured HUVEC or mixtures of
endothelial cells (EC) and B lymphoblastoid cells (BLC) induce the
differentiation of purified CD8+ PBL into allospecific,
class I MHC-restricted CTL that lyse EC, but not BLC autologous to EC.
Furthermore, these EC-selective CTL lines secrete little IFN-
after
target cell contact. In the present study, we have analyzed these
polyclonal populations at a single cell level by cloning at limiting
dilution and propagating the resulting CTL clones in the absence of EC.
Phenotypically stable, alloreactive EC-selective CTL preferentially
emerge from cocultures in which EC or EC + BLC are the initial
stimulating cell types compared with cocultures stimulated by BLC alone
(p = 0.005). Compared with BLC-stimulated CTL,
EC-stimulated CTL clones often fail to secrete IFN- after target
cell contact (p = 0.0006) and constitutively
express CD40 ligand (CD40L) at rest (p = 0.0006).
The absence of IFN- secretion does not result from a switch to IL-4
secretion. The expression of CD40L inversely correlates with the
secretion of IFN- after target cell contact (p =
0.0001), but correlations of CD40L expression and failure to secrete
IFN- with EC-selective killing did not reach statistical
significance. We conclude that in a microenvironment in which
allogeneic EC are in close contact with infiltrating CD8+ T
cells, such as within a graft arterial intima, CTL subsets may emerge
that display EC selectivity or express CD40L and secrete little IFN-
after Ag contact.
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