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Blood, Vol. 93 No. 11 (June 1), 1999:
pp. 3876-3884
Interleukin-2-Activated Rat Natural Killer Cells Express Inducible
Nitric Oxide Synthase That Contributes to Cytotoxic Function and
Interferon- Production
M. Grazia Cifone,
Simona D'Alò,
Raffaella Parroni,
Danilo Millimaggi,
Leda Biordi,
Stefano Martinotti, and
Angela Santoni
From the Department of Experimental Medicine, University of
L'Aquila, L'Aquila; and the Department of Experimental Medicine and
Pathology, University La Sapienza, Rome, Italy.
Natural killer (NK) cells are large granular lymphocytes capable of
destroying cells infected by virus or bacteria and susceptible tumor
cells without prior sensitization and restriction by major histocompatability complex (MHC) antigens. Their cytotoxic activity could be strongly enhanced by interleukin-2 (IL-2). Previous findings, even if obtained with indirect experimental approaches, have suggested a possible involvement of the inducible nitric oxide (iNOS) pathway in
the NK-mediated target cell killing. The aim of the present study was
first to directly examine the induction of iNOS in IL-2-activated rat
NK cells isolated from peripheral blood (PB-NK) or spleen (S-NK), and
second to investigate the involvement of the iNOS-derived NO in the
cytotoxic function of these cells. Our findings clearly indicate the
induction of iNOS expression in IL-2-activated PB-NK and S-NK cells,
as evaluated either at mRNA and protein levels. Accordingly,
significantly high levels of iNOS activity were shown, as detected by
the L-arginine to L-citrulline conversion in appropriate assay
conditions. The consequent NO generation appears to partially account
for NK cell-mediated DNA fragmentation and lysis of sensitive tumor
target cells. In fact, functional inhibition of iNOS through specific
inhibitors, as well as the almost complete abrogation of its expression
through a specific iNOS mRNA oligodeoxynucleotide antisense,
significantly reduced the lytic activity of IL-2-activated NK
cells. Moreover, IL-2-induced interferon- production appears also to be dependent, at least in part, on iNOS induction.
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