Surface-displayed glyceraldehyde 3-phosphate dehydrogenase and galectin from Dirofilaria immitis enhance the activation of the fibrinolytic system of the host

Acta Trop. 2015 May;145:8-16. doi: 10.1016/j.actatropica.2015.01.010. Epub 2015 Feb 7.

Abstract

Cardiopulmonary dirofilariosis is a cosmopolitan disease caused by Dirofilaria immitis, a filaroid parasite whose adult worms live for years in the vascular system of its host. Previous studies have shown that D. immitis can use their excretory/secretory (ES) and surface antigens to enhance fibrinolysis, which could limit the formation of clots in its surrounding environment. Moreover, several isoforms of the glyceraldehyde 3-phosphate dehydrogenase (GAPDH) and galectin (GAL) were identified in both antigenic extracts as plasminogen-binding proteins. The aim of this work is to study the interaction of the GAPDH and GAL of D. immitis with the fibrinolytic system of the host. This study includes the cloning, sequencing and expression of the recombinant forms of the GAPDH and GAL of D. immitis (rDiGAPDH and rDiGAL) and the analysis of their capacity as plasminogen-binding proteins. The results indicate that rDiGAPDH and rDiGAL are able to bind plasminogen and stimulate plasmin generation by tissue plasminogen activator (tPA). This interaction needs the involvement of lysine residues, many of which are located externally in both proteins as have been shown by the molecular modeling of their secondary structures. In addition, we show that rDiGAPDH and rDiGAL enhance the expression of the urokinase-type plasminogen activator (uPA) on canine endothelial cells in culture and that both proteins are expressed on the surface of D. immitis in close contact with the blood of the host. These data suggest that D. immitis could use the associated surface GAPDH and GAL as physiological plasminogen receptors to shift the fibrinolytic balance towards the generation of plasmin, which might constitute a survival mechanism to avoid the clot formation in its intravascular habitat.

Keywords: Dirofilaria immitis; Fibrinolysis; Galectin; Glyceraldehyde 3-phosphate dehydrogenase; Plasminogen.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Animals
  • Antigens, Surface / metabolism*
  • Dirofilaria immitis / enzymology*
  • Dogs / parasitology*
  • Fibrinolysin / metabolism
  • Fibrinolysis / physiology*
  • Galectins / metabolism*
  • Glyceraldehyde 3-Phosphate / metabolism*
  • Helminth Proteins / genetics
  • Helminth Proteins / metabolism
  • Host-Parasite Interactions / physiology*
  • Humans
  • Plasminogen / metabolism

Substances

  • Antigens, Surface
  • Galectins
  • Helminth Proteins
  • Glyceraldehyde 3-Phosphate
  • Plasminogen
  • Fibrinolysin