PERTUSSIS TOXIN FROM BORDETELLA-PERTUSSIS BLOCKS NEUTROPHIL MIGRATIONAND NEUTROPHIL-DEPENDENT EDEMA IN RESPONSE TO INFLAMMATION

Pertussis toxin (Ptx) is a hexameric protein with classical AB archite cture produced by Bordetella pertussis. The aim of this study was to i nvestigate the effect of Ptx on migration of polymorphonuclear leukocy tes to site of inflammation and on cell-dependent edema. Ptx was purif ied from the supernatant of the culture medium of B. pertussis using h ydroxylapatite chromatography and fetuin affinity chromatography. Ptx induced a maximal clustering of Chinese hamster ovary cells at concent rations as low as 0.1 ng/ml. Intravenous injection of Ptx (400 ng) sig nificantly blocked the neutrophil migration induced by 200 ng of lipop olysaccharide (LPS from E. coli O111:B4; 2.27 +/- 0.13 vs 0.61 +/- 0.1 6 per 10(6) neutrophils/ml; P<0.001; N = 5) and by 200 ng of formyl-me thionyl-leucyl-phenylalanine (fMLP; 2.53 +/- 0.45 vs 0.75 +/- 0.14 per 10(6) neutrophils/ml; P<0.01; N = 6) into the peritoneal cavities of male Wistar rats (weighing 150-180 g). In addition, Ptx (400 ng) pretr eatment also blocked the edema induced by intraplantar injection of 10 0 mu g carrageenin (Delta increase in volume: 0.667 +/- 0.087 vs 0.313 +/- 0.058 ml; P<0.01; N = 5) but not the edema induced by 100 mu g de xtran (Delta increase in volume: 0.537 +/- 0.06 vs 0.385 +/- 0.076 ml; P>0.05; N = 5). These data demonstrate that Ptx blocked neutrophil mi gration induced by a direct fMLP stimulus of a site of inflammation. I n addition, this toxin blocks the indirect stimulus of LPS on neutroph il migration. Furthermore, Ptx also inhibits the neutrophil-dependent edema induced by carrageenin, but not the edema induced by dextran tha t is in part dependent on basophil cells. These results warrant furthe r studies on the mechanisms of Ptx inhibition of neutrophil-dependent edema and cell migration.