High-molecular-weight kininogen fragments stimulate the secretion of cytokines and chemokines through uPAR, Mac-1, and gC1qR in monocytes

MM Khan, HN Bradford, I Isordia-Salas… - … , and vascular biology, 2006 - Am Heart Assoc
MM Khan, HN Bradford, I Isordia-Salas, Y Liu, Y Wu, RG Espinola, B Ghebrehiwet…
Arteriosclerosis, thrombosis, and vascular biology, 2006Am Heart Assoc
Objective—Plasma high-molecular-weight kininogen (HK) is cleaved in inflammatory
diseases by kallikrein to HKa with release of bradykinin (BK). We postulated a direct link
between HKa and cytokine/chemokine release. Methods and Results—HKa, but not BK,
releases cytokines tumor necrosis factor (TNF)-α, interleukin (IL)-1β, IL-6, and chemokines IL-
8 and MCP-1 from isolated human mononuclear cells. At a concentration of 600 nM,
glutathione-S-transferase (GST) fusion proteins of kininogen domain 3 (D3), a fragment of …
Objective— Plasma high-molecular-weight kininogen (HK) is cleaved in inflammatory diseases by kallikrein to HKa with release of bradykinin (BK). We postulated a direct link between HKa and cytokine/chemokine release.
Methods and Results— HKa, but not BK, releases cytokines tumor necrosis factor (TNF)-α, interleukin (IL)-1β, IL-6, and chemokines IL-8 and MCP-1 from isolated human mononuclear cells. At a concentration of 600 nM, glutathione-S-transferase (GST) fusion proteins of kininogen domain 3 (D3), a fragment of domain 3, E7P (aaG255-Q292), HK domain 5 (D5), the D5 recombinant peptides HG (aa K420-D474) and HGK (aa H475-S626) stimulated secretion of IL-1β from mononuclear cells. Monoclonal antibodies (MAbs) specific for D5 or specific for D3 blocked release of IL-1β by HKa, supporting the importance of both domains. Antibodies to HK receptors on leukocytes including Mac-1, LFA-1, uPAR, and C1qR inhibited IL-1β secretion induced by tKa 98%, 89%, 85%, and 62%, respectively. Fractionation of mononuclear cells identified the responsible cell, a blood monocyte. Inhibitors of signaling pathways NFkB, JNK, and p38 but not extracellular signal-regulated kinase (ERK) decreased cytokine release from mononuclear cells. HKa increased the synthesis of IL-1β as deduced by an increase of IL-1β mRNA at 1 to 2 hours.
Conclusions— HKa domains 3 and 5 may contribute to the pathogenesis of inflammatory diseases by releasing IL-1β from human monocytes using intracellular signaling pathways initiated by uPAR, β2 integrins and gC1qR.
Am Heart Assoc