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Blood, Vol. 95 No. 6 (March 15), 2000:
pp. 2068-2075
Inducible loss of NF- B activity is associated with apoptosis
and Bcl-2 down-regulation in Epstein-Barr virus-transformed B
lymphocytes
Jean Feuillard,
Marino Schuhmacher,
Sylvie Kohanna,
Marianne Asso-Bonnet,
Frédérique Ledeur,
Raymonde Joubert-Caron,
Philippe Bissières,
Axel Polack,
Georg W. Bornkamm, and
Martine Raphaël
From Biochimie Cellulaire des Hémopathies Lymphoïdes,
Université Paris, Bobigny, France, and Institut für
Klinische Molekularebiologie and Tumorgenetik, München, Germany.
The Epstein-Barr virus (EBV)-encoded latent membrane protein-1
induces NF- B activity by targeting IB . To
understand the role of NF-B activation in EBV-related oncogenesis,
we have subcloned mutated IB32/36A cDNA into a
pHEBo vector containing doxycycline regulatory sequences and stably
transfected this construct into a lymphoblastoid cell line. Two tightly
regulated clones were obtained in which IB32/36A
was inducible in a doxycycline dose-dependent manner. Levels of
inducible IB32/36A peaked at day 2. Inhibition of
NF-B activity was closely correlated with levels of inducible IB32/36A. Levels of 3 well-known
NF-B-dependent genes, CD54, p105, and endogenous IB, were
decreased when IB32/36A was induced, and
the growth of IB32/36A-induced EBV-infected cells was slightly reduced. Loss of NF-B activity was associated with decreased Bcl-2 protein levels. Finally, the induction of apoptosis was
strongly increased in IB32/36A-overexpressing
cells. Together these results show that it is possible to control
IB32/36A levels, ie, NF-B activity, in
EBV-infected B-lymphocytes using a doxycycline-inducible vector.
Moreover, our results indicate that NF-B can protect EBV-infected
cells from apoptosis by Bcl-2. Finally, our results suggest that a
cellular model with doxycycline-inducible IB32/36A may be useful in the identification of genuine NF-B target genes in EBV-infected B cells.
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