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Program in Molecular Cardiobiology, Boyer Center for Molecular Medicine, Yale University School of Medicine, New Haven, CT 06510
We compared the capacity of cultured human endothelial cells (EC)
vs B lymphoblastoid cells (BLC) from the same donor to stimulate
allogeneic CD8+ T cells to differentiate into CTL, assaying
for allorestricted cytotoxicity, T cell growth, IFN-
secretion, and
perforin expression. The input cell number affected specificity and
potency of the resulting CTL. At low input (<105
cells/well), anti-EC CTL were rarely detected. At high input
(>106 cells/well), anti-EC CTL developed that
displayed unrestricted, low-titer killing and an unstable phenotype. At
intermediate input (1.0–2.5 x 105 cells/well),
classical class I MHC-restricted, CD8+, and
perforin-positive anti-EC CTL developed with reproducible
frequencies. However, under all conditions EC were less efficient
stimulators than BLC from the same donor. Anti-EC CTL did not kill BLC,
whereas anti-BLC CTL killed BLC and EC from the same donor with
comparable efficiency. When CD8+ T lymphocytes were grown
in the presence of EC and BLC together, the differentiation of
anti-BLC CTL was completely suppressed, while the anti-EC
response was intact. The inhibition of the allogeneic anti-BLC CTL
response was independent of T cell-EC contact, and proliferation of
CD8+ T cells was inhibited by EC-conditioned medium. We
conclude that EC are competent but less efficient activators of CTL
differentiation than are BLC and that EC actively regulate
differentiation and/or expansion of allospecific
CTL.
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