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Platelet-Activating Factor Synthesized by IL-12-Stimulated Polymorphonuclear Neutrophils and NK Cells Mediates Chemotaxis¹

Benedetta Bussolati^*,, Filippo Mariano^*, Alessandro Cignetti, Anna Guarini, Vincenzo Cambi, Robert Foà, Giuseppe Piccoli^* and Giovanni Camussi^2,§

^* Laboratorio di Immunopatologia Renale, Dipartimento di Discipline Medico-Chirurgiche, Dipartimento di Scienze Biomediche ed Oncologia Umana Università di Torino; and Cattedra di Nefrologia, Università di Parma, and ^§ Dipartimento di Scienze Cliniche e Biologiche II Facoltà di Medicina, Università di Pavia, Varese, Italy

IL-12 is chemotactic for NK cells and polymorphonuclear neutrophils (PMN), but not for monocytes. In the present study, we evaluated whether the chemotactic effect of IL-12 is a direct phenomenon or is dependent on the generation of secondary mediators. The results obtained indicate that IL-12 induces a dose- and time-dependent synthesis of platelet-activating factor (PAF) from PMN and NK cells and of reactive oxygen radicals (ROS) from PMN. Monocytes and CD56-negative PBMC cells did not synthesize PAF or ROS after challenge with IL-12. The production of ROS by PMN was significantly inhibited by two chemically different PAF receptor antagonists (WEB 2170 and CV 3988), suggesting an autocrine stimulation of PMN by PAF newly synthesized after the challenge with IL-12. Moreover, the IL-12-induced chemotaxis of PMN and NK cells was significantly reduced by both WEB 2170 and CV 3988, suggesting that synthesized PAF mediates the chemotactic effect of IL-12. Preincubation with superoxide dismutase, which blocks the formation of superoxide anions, also reduced the chemotactic effect of IL-12 on PMN, but not on NK cells, suggesting that superoxide anion generation is relevant only for the IL-12-induced chemotaxis of PMN. In conclusion, the results of the present study indicate that IL-12-induced PAF synthesis plays a critical role in triggering the events involved in the motogenic response of PMN and NK to IL-12.

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