Antineutrophil cytoplasmic antibodies induce monocyte IL-8 release - Role of surface proteinase-3, alpha 1-antitrypsin, and Fc gamma receptors

Cytoplasmic antineutrophil cytoplasmic antibodies (cANCA) that accompany the neutrophilic vasculitis seen in Wegener's granulomatosis (WG), are directed against proteinase-3 (PR-3), a serine proteinase which is located in azurophilic granules of neutrophils and monocytes. PR-3, when expressed on the surface of TNF alpha-primed neutrophils, can directly activate neutrophils by complexing cANCA and promoting concomitant Fc gamma receptor (Fc gamma R) cross-linking, Although the neutrophil's pathogenic role in WG has been studied, the role of the monocyte has not been explored, The monocyte, with its ability to release cytokines and regulate neutrophil influx, also expresses PR-3. Therefore, the monocyte may play a significant role in WG via the interaction of surface PR-3 with cANCA, inducing cytokine release by the monocyte, To test this hypothesis, monocytes were studied for PR-3 expression and for IL-8 release in response to cANCA IgG, PBMC obtained from healthy donors displayed dramatic surface PR-3 expression as detected by immunohistochemistry and flow cytometry in response to 0.5-h pulse with TNF alpha (2 ng/ml), Purified monoclonal anti-PR-3 IgG added to TNF alpha-primed PBMC induced 45-fold more IL-8 release than an isotype control antibody, Furthermore, alpha l-antitrypsin (alpha 1-AT), the primary PR-3 antiprotease, inhibited the anti-PR-3 induced IL-8 release by 80%, Importantly, Fab and F(ab')(2) fragments of anti-PR-3 IgG, which do not result in Fc gamma receptor cross-linking, do not induce IL-8 release, As a correlate, IgG isolated from cANCA positive patients with WG induced six times as much PBMC IL-8 release as compared to IgG isolated from normal healthy volunteers. Consistent with PR-3 associated IL-8 induction, alpha 1-AT significantly inhibited this effect, These observations suggest that cANCA may recruit and target neutrophils through promoting monocyte IL-8 release, This induction is mediated via Fc gamma receptor cross-linking and is regulated in part by alpha 1-AT.