Inhibition of chemotaxis Ng-monomethyl-L-arginine: a role for cyclic GMP

Abstract

The metabolism of L-arginine to nitric oxide (NO) has been shown to be important for the effector functions of many cell types, including polymorphonuclear (PMN) leukocytes. Its effect appears to be mediated at least in part by NO stimulation of soluble guanylate cyclase. We evaluated the role of this pathway in two PMN effector functions: cell movement and microbial killing, using the competitive inhibitor of L- arginine conversion to NO, NG-monomethyl-L-arginine (NMA). We also evaluated the effect of additional L-arginine and dibutyryl cyclic guanosine monophosphate (cGMP) on any NMA-associated changes. Human peripheral blood neutrophils were used and the cells were incubated with and without NMA. Chemotaxis was evaluated using a 48-well micro- Boyden chamber. Microbial killing was evaluated using S aureus strains D2C and 502A. These studies demonstrated that chemotaxis to formyl- methionyl-leucyl-phenylalanine was markedly inhibited in NMA-treated cells. This inhibition could be overcome if L-arginine or dibutyryl cGMP were added with the NMA. In contrast, microbial killing of S aureus was unaffected by NMA. These observations support the hypothesis that the L-arginine metabolism to NO and its effect on the cGMP level may be important for the dynamic changes required for neutrophil chemotaxis.

Role of cyclic GMP on inhibition by nitric oxide donors of human eosinophil chemotaxis in vitro

“…The use of different nitric oxide synthase (NOS) inhibitors or NO donors has indicated that endogenous NO may limit ( Kubes et al , 1991 ; Gaboury et al , 1993 ; Kurose et al , 1994 ,1995) or potentiate ( Ferreira et al , 1996 ,1998; Feder et al , 1997 ) the leukocyte adhesion and/or recruitment into normal or inflamed microcirculation. Stimulation or attenuation of leukocyte activity has also been described in isolated cells with the use of NOS inhibitors or NO donors ( Kaplan et al , 1989 ; Belenky et al , 1993a ,1993b; Moilanen et al , 1993 ; Beauvais et al , 1995 ; Zanardo et al , 1997 ; Okayama et al , 1998 ; Kosonen et al , 1999 ; Thomazzi et al , 2001 ). Although these discrepancies have not been clarified yet, it has been suggested that variations in cGMP levels may explain these opposite effects.…”

“…Although cyclic nucleotides are known to modulate leukocyte activation processes, their role in cell locomotion is still controversial. In human isolated neutrophils and mononuclear cells, increase of cGMP levels has been shown to either stimulate ( Kaplan et al , 1989 ; Belenky et al , 1993a ,1993b; Wanikiat et al , 1997 ) or inhibit ( Schröder et al , 1990 ; Bath et al , 1991 ; Moilanen et al , 1993 ; Kosonen et al , 1999 ; Conran et al , 2001 ) the in vitro chemotaxis and adhesion. On the other hand, the soluble guanylyl cyclase inhibitor 1 H ‐[1,2,4] oxidiazolo[4,3‐ α ] quinoxalin‐1‐one (ODQ) has been clearly shown to reduce concentration‐dependently N ‐formyl‐ L ‐methionyl‐ L ‐leucyl‐phenylalanine (fMLP)‐induced eosinophil chemotaxis, suggesting a key role for cGMP in these cells ( Zanardo et al , 1997 ; Thomazzi et al , 2001 ).…”

Hemin, a heme oxygenase substrate analog, inhibits the cell surface expression of CD11b and CD66b on human neutrophils

“…Studies suggest that NO, derived from an inducible isoform of NO synthase, may act as an endogenous modulator of the inflammatory response by inhibiting adhesion of leukocytes to endothelial cells (41–43). It is also known that exogenous NO facilitates neutrophil chemotaxis in vitro (44) and that inhibition of NO synthase decreases neutrophil migration in vitro (45, 46). Furthermore, the addition of NO to the inhaled air reduces the cell surface expression of CD11/CD18 on circulating neutrophils, whereas the incubation of isolated neutrophils with L‐NMMA, an inhibitor of NO synthase, increases their expression of CD11/CD18 (47, 48).…”

Nitric oxide‐dependent neutrophil recruitment: role in nasal secretion

“…Recent studies suggest that NO derived from an inducible isoform of NO synthase may act as an endogenous modulator of the inflammatory response by inhibiting adhesion of leucocytes to endothelial cells [13–15]. It is also known that exogenous NO facilitates neutrophil chemotaxis in vitro [9], and that inhibition of NO synthase decreases neutrophil migration in vitro [10,11]. Here, it is shown that L‐NAME markedly inhibits LTB4‐induced neutrophil‐dependent nasal secretion in vivo .…”

“…The nasal production of NO is reported to increase during inflammation and allergic rhinitis due to increased activity of an inducible isoform of NO [6–8]. Exogenous NO has been shown to facilitate neutrophil chemotaxis in vitro [9], whereas inhibition of NO prevents neutrophil recruitment [10,11]. Therefore, it was hypothesized that inhibition of NO in the nose, by preventing neutrophil recruitment, would prevent neutrophil‐dependent nasal secretion.…”