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Blood, Vol. 92 No. 6 (September 15), 1998:
pp. 2093-2102
The Induction of Nitric Oxide by Interleukin-12 and Tumor Necrosis
Factor- in Human Natural Killer Cells: Relationship With the
Regulation of Lytic Activity
Ombretta Salvucci,
Jean Pierre Kolb,
Bernard Dugas,
Nathalie Dugas, and
Salem Chouaib
From the Laboratoire Cytokines et Immunologie des Tumeurs Humaines, U
487 INSERM, Institut Gustave-Roussy, Villejuif, France; the
Interférons et cytokines, Unité 365 INSERM, Institut Curie,
Paris, France; the Laboratoire d'Immuno-Hématologie, CNRS URA
625, Hôpital la Pitié Salpétrière, Paris,
France; and the Laboratoire Virus Neurone et Immunité, IFR
Kremlin Bicètre, Kremlin Bicètre, France.
We have investigated the interleukin-12 (IL-12) and tumor necrosis
factor- (TNF)-induced regulation of human natural killer (NK)
cell function and their relationship with nitric oxide (NO) generation.
We demonstrate that both cytokines were efficient to trigger the
transcription of the inducible nitric oxide synthase (iNOS) mRNA, as
detected by reverse transcriptase-polymerase chain reaction (RT-PCR).
Western blot analysis and intracytoplasmic fluorescence showed that
iNOS protein was also induced by both cytokines. However, our data
indicate that NO does not play a significant role in the effector phase
of the cytotoxic activity mediated by NK-stimulated cells, inasmuch as
the lytic activity was not affected in the presence of specific NO
synthase inhibitors. When aminoguanidine (AMG), an inhibitor of iNOS,
was added during the afferent phase of NK stimulation with IL-12 and
TNF, a subsequent increase in the lytic potential of the effector
cells towards the NK-sensitive target cells (K562) and
lymphokine-activated killer (LAK) target cells (Daudi) was observed.
Conversely, the addition of chemical NO donors during the afferent step
resulted in a dose-dependent inhibition of the NK and LAK cytotoxicity. Our data suggest that the enhancement of NK-cell cytotoxic activity resulting from iNOS inhibition may be correlated, at least in part, to
an increase in interferon- production and granzyme B expression.
© 1998 by The American Society of Hematology.
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