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Infection and Immunity, September 1998, p. 4068-4072, Vol. 66, No. 9
0019-9567/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Murine Macrophages Use Oxygen- and Nitric Oxide-Dependent Mechanisms To Synthesize S-Nitroso-Albumin and To Kill Extracellular Trypanosomes

Alain P. Gobert,1,2 Silla Semballa,1 Sylvie Daulouede,1 Sophie Lesthelle,1 Murielle Taxile,2 Bernard Veyret,2 and Philippe Vincendeau1,*

Laboratoire de Parasitologie, Université de Bordeaux II, Bordeaux,1 and Laboratoire de Physique des Interactions Ondes-Matières, UMR 5501, CNRS, ENSCPB, Talence,2 France

Received 17 February 1998/Returned for modification 1 April 1998/Accepted 5 June 1998

Reactive nitrogen intermediates were synthesized spontaneously in cultures of macrophages from Trypanosoma brucei brucei-infected mice by an inducible nitric oxide (NO) synthase. This was inhibited by the addition of nitro-L-arginine. In this paper, we report the kinetics of the fixation of macrophage-derived NO on bovine serum albumin by using an enzyme-linked immunosorbent assay. S nitrosylation was confirmed by the Saville reaction, using mercuric chloride. It is known that reactive oxygen intermediates (ROI) are also synthesized by stimulated macrophages. The fact that NO is able to bind cysteine only under aerobic conditions led us to investigate the role of macrophage-derived ROI in the formation of S-nitrosylated proteins by activated macrophages. The immunoenzymatic signal decreased by 66 and 30% when superoxide dismutase and catalase, respectively, were added to the culture medium of macrophages from infected mice. In addition, the decrease in S-nitrosylated albumin formation correlated with the protection of extracellular trypanosomes from the cytostatic and cytotoxic activity of NO. Melatonin, a hydroxyl radical scavenger resulting from the decomposition of peroxynitrous acid, had no effect. All these data support the concept that an interaction between NO and ROI promoted the production of S-nitroso-albumin by activated macrophages from infected mice.

* Corresponding author. Mailing address: Laboratoire de Parasitologie, Université de Bordeaux II, Bat 1B, 146 rue Léo Saignat, 33076 Bordeaux Cédex, France. Phone: 33-557-57-17-73. Fax: 33-556-84-66-31. E-mail: Philippe.Vincendeau{at}parasito.u-bordeaux2.fr.

Infection and Immunity, September 1998, p. 4068-4072, Vol. 66, No. 9
0019-9567/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


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