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B Cascade Is Important in Bcl-xL Expression and for the Anti-Apoptotic Effects of the CD28 Receptor in Primary Human CD4+ Lymphocytes1
*
Division of Clinical Immunology and Allergy, Department of Medicine, University of California, Los Angeles, CA 90095; and
Signal Pharmaceuticals, San Diego, CA 92121
We explored the role of the NF-
B pathway in the survival of
primary human CD4+ T lymphocytes during CD28 costimulation.
Transduction of proliferating CD4+ T cells with a
tetracycline-regulated retrovirus encoding for a dominant-interfering,
degradation-resistant I-B
(inhibitor of B factor) mutant
induced apoptosis. Using DNA arrays, we show that Bcl-xL
features as a prominent anti-apoptotic member among a number of
early CD28-inducible genes. A 1.2-kb segment of the proximal
Bcl-xL promoter, linked to a luciferase reporter, responded
to CD3/CD28 stimulation in Jurkat cells. Mutation of an NF-B site
around -840 decreased, while ectopic expression of I-B kinase-ß
(IKKß) enhanced reporter gene activity. Na+-salicylate
and cyclopentenone PGs, direct inhibitors of IKKß, interfered in the
activation of the Bcl-xL promoter and induced apoptosis in
CD28-costimulated CD4+ T cells. Moreover, salicylate
blocked nuclear localization of NF-B factors that bind to the
NF-B binding site in the Bcl-xL promoter, as well as the
expression of Bcl-xL protein. HuT-78, a lymphoblastoid T
cell line with constitutive NF-B activity, contained elevated levels
of Bcl-xL protein and, similar to proliferating
CD4+ T cells, was resistant to apoptotic stimuli such as
anti-Fas and TNF-. In contrast, the same stimuli readily induced
apoptosis in a Jurkat T cell clone with no detectable
Bcl-xL expression. Jurkat BMS2 cells also differed from
HuT-78 in collapse of mitochondrial membrane potential and superoxide
generation in the mitochondrium. Taken together, these data demonstrate
that CD3/CD28-induced activation of IKKß and expression of
Bcl-xL promote the survival of primary human
CD4+ T lymphocytes.
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