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Las brucelas son bacterias intracelulares facultativas, capaces de sobrevivir y multiplicarse en el interior de diversas celulas, incluyendo los macrofagos. Su virulencia y tendencia a producir infecciones cronicas se cree que es debida a su capacidad para resistir los mecanismos bactericidas intracelulares, entre los que se encuentra la produccion de oxido nitrico (NO). Teniendo en cuenta que el lipopolisacarido liso (SLPS) de las brucelas ha sido propuesto como factor de virulencia de la mismas y que la mayoria de efectos biologicos del LPS se asocian al lipido A (LipA), nos planteamos como objetivo de esta tesis comprobar si los S-LPS y LipA de Brucella inducian la produccion de NO en los macrofagos peritoneales de rata, comparando su efecto con el del LPS y LipA de Escherichia coli. = We show in this study that smooth lipopolysaccharide (S-LPS) of Brucella abortus 544 and Brucella melitensis 16 M are able to induce nitric oxide (NO) production in rat peritoneal macrophages in a time- and dose-dependent manner. The involvement of the L-arginine pathway is supported by the dependence of this production on the presence of L-arginine and the inhibition by NG-metil-L-arginina, an inhibitor of NO synthases. Evidence in favour of the involvement of the inducible isoform of the NO synthase (iNOS) are the increased expression of both iNOS mRNA and iNOS protein elicited by both S-LPS. The lipid A (lipA) moieties of S-LPS also induced the expression of iNOS mRNA and NO production, thus suggesting that these moieties could be the portion of the S-LPS molecule involved in NO induction. The amounts of NO produced by either S-LPS reached similar values, as well as those elicited by lipA, however, they were less active than their counterparts from Escherichia coli at the range of concentrations

Inducción de la producción de óxido nítrico por lipopolisacárido de Brucella en macrófagos de rata