Myxoma viral serpin, Serp-1, a unique interceptor of coagulation and innate immune pathways.
Serpins maintain haemostasis through regulation of serine proteinases in the thrombotic and thrombolytic pathways. Viruses encode serpins that can alter thrombotic and thrombolytic responses producing, in some cases, disseminated intravascular coagulation (DIC). However, it has not been precisely defined how viral serpins induce these profound responses. The rabbit myxoma viral serpin, Serp-1 inhibits urokinase- and tissue-type plasminogen activators (uPA and tPA), plasmin and factor Xa in vitro and exhibits remarkable anti-inflammatory activity in various animal models. The effects of Serp-1 on activation of human platelets, endothelial cells, monocytes and T cells that mediate thrombosis and innate immune responses were therefore examined. We found that Serp-1 attenuated platelet and mononuclear cell adhesion to fibronectin and collagen. Serp-1 similarly inhibited monocyte migration into the peritoneum. Serp-1 inhibition of monocyte migration was lost in uPA receptor (uPAR) deficient mice. Serp-1 bound to the plasma membrane surface and altered uPA activation of endothelial cells (p=0.001), thrombin activation of platelets (p=0.021) and phorbol ester activation of endothelial (p=0.047), monocyte (p=0.011) and Jurkat T cells (p=0.012) as measured by intracellular calcium. Modulation of cellular activation was confirmed by membrane fluidity analysis. Microarray analysis of Serp-1 treated endothelial cells revealed alterations in Inositol 1,4,5-triphosphate receptor type II (ITPR2) a calcium-regulating gene. This study demonstrates the unique capacity of a viral serpin, Serp-1 to modify adhesion, activation, gene expression and calcium homeostasis in a wide range of cells that regulate coagulation and inflammation. Endothelial cells potentially represent a pivotal regulatory point for Serp-1 anti-inflammatory activity.
Viswanathan
Kasinath
K
Vascular Biology Research Group, Robarts Research Institute, 100 Perth Drive, P.O. Box 5015, London, ON, N6A 2K8, Canada.
Liu
Liying
L
Vaziri
Sepideh
S
Dai
Erbin
E
Richardson
Jakob
J
Togonu-Bickersteth
Babajide
B
Vatsya
Pracha
P
Christov
Alexander
A
Lucas
Alexandra R
AR
D000818:Animals; D000893:Anti-Inflammatory Agents / Q000494:pharmacology*; D001777:Blood Coagulation / Q000187:drug effects*; D001792:Blood Platelets / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure; D002118:Calcium / Q000378:metabolism; D015220:Calcium Channels / Q000235:genetics / Q000378:metabolism; D002448:Cell Adhesion; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism; D002465:Cell Movement; D042783:Endothelial Cells / Q000187:drug effects* / Q000378:metabolism / Q000648:ultrastructure; D005343:Fibrinolytic Agents / Q000494:pharmacology*; D005786:Gene Expression Regulation; D006801:Humans; D007113:Immunity, Innate / Q000187:drug effects; D053496:Inositol 1,4,5-Trisphosphate Receptors; D019169:Jurkat Cells; D008560:Membrane Fluidity; D051379:Mice; D009000:Monocytes / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure; D010703:Phorbol Esters / Q000494:pharmacology; D010973:Platelet Adhesiveness; D011480:Protease Inhibitors / Q000494:pharmacology; D018160:Receptors, Cytoplasmic and Nuclear / Q000235:genetics / Q000378:metabolism; D015843:Serpins / Q000494:pharmacology*; D013601:T-Lymphocytes / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure; D013917:Thrombin / Q000494:pharmacology; D014568:Urokinase-Type Plasminogen Activator / Q000494:pharmacology; D014764:Viral Proteins / Q000494:pharmacology*
D000893:Anti-Inflammatory Agents; D015220:Calcium Channels; D005343:Fibrinolytic Agents; C506926:ITPR1 protein, human; C506927:ITPR2 protein, human; D053496:Inositol 1,4,5-Trisphosphate Receptors; D010703:Phorbol Esters; D011480:Protease Inhibitors; D018160:Receptors, Cytoplasmic and Nuclear; D015843:Serpins; D014764:Viral Proteins; D013917:Thrombin; D014568:Urokinase-Type Plasminogen Activator; D002118:Calcium