Andras Gruber Laboratory

Summary:

Current Research Goals:
To determine the role of contact activation in acute intraluminal thrombus propagation using synthetic vascular grafts in a primate model of thrombosis and hemostasis. If the functionality of the contact system enzyme complex (FXI/FXII/KK/HMWK) is relevant to the pathogenesis of thrombosis, a FXI inhibitor could become the first safe antithrombotic agent.

To characterize the effects of endogenous protein C activation on acute arterial thrombogenesis and hemostasis using rationally engineered recombinant enzymes. A pharmacologically viable protein C activator could help utilize the body's own antithrombotic and antiinflammatory system similar to the way streptokinase and tPA became useful fibrinolysis activators.

Affiliations:

• Department of Biomedical Engineering

People:

• PI: Gruber, Andras, M.D.
  

Resources:

Biological Specimens

• Baboon plasma ( Biological Specimen )
  

  Baboon blood was collected into 3.2% sodium citrate anticoagulant (9:1, v/v). Platelet-rich plasma was prepared and frozen at −80°C.

• C57B/6 murine plasma ( Biological Specimen )
  

• FXI +/+ murine plasma samples ( Biological Specimen )
  

• FXI-/- murine plasma sample ( Biological Specimen )
  

Instruments

• Electromechanical fibrometer ( Fibrometer )
  

• GE-400T gamma scintillation camera ( Gamma camera )
  

  Interfaced with a NuQuest InteCam computer system.

• Hemavet HV950FS analyzer ( Hematology analyzer )
  

  "The HEMAVET® provides superior cell separation for accurate and precise analysis through use of patented Focused Flow Technology (eleven patents issued) thus giving the best technology available for dealing with veterinary samples. In addition patented Expectation Maximization software helps categorize the different types of cells."

• Molecular Devices SpectroMax 340 microplate reader ( Microplate reader )
  

  The SpectroMax 340 "mimics a dual beam spectrophotometer, measures each sample in the plate directly, and eliminates measurement error due to variations in light output between optical fibers on other instruments for more reliable data reporting. The eight-channel system delivers both high precision and speed when reading 96- and 384-well microplates for endpoint measurements, kinetic assays, and spectral scans."

• Serono System 9000 Hematology Series cell counter ( Automatic cytometer )
  

Organisms and Viruses

• C57BL/6 mice ( Mus musculus )
  

• FXI+/+ mice ( Mus musculus )
  

• FXI-/- mice ( Mus musculus )
  

  "The FXI-/- genotype was backcrossed onto the C57BL/6 background for >9 generations."

• Papio anubis ( Papio anubis )
  

Protocols

• Murine arterial thrombosis model ( Protocol )
  

  Model used to study thrombosis formation in mouse models. Thrombus formation induced by ferric chloride or laser injury.

• Murine peritoneal sepsis model ( Protocol )
  

  Model is used to study the role of anticoagulation in the treatment of sepsis-associated consumptive coagulopathy.

Reagents

• Neutralizing monoclonal IgG 14E11 to mammalian factor XI ( Monoclonal antibody reagent )
  

  "Clone 14E11 was expanded in a CL1000 bioreactor (Integra Biosciences), and immunoglobulin G (IgG) was purified by cation exchange and thiophilic agarose chromatography. 14E11 was biotinylated using an EZ-link sulfo-NHS-biotinylation kit (Thermo Scientific)."

• Neutralizing monoclonal IgG O1A6 to human factor XI ( Monoclonal antibody reagent )
  

  Antibody binds to the A3 domain of human fXI and interferes with activation of factor IX by fXIa.

Web Links:

• http://www.ohsu.edu/xd/education/schools/school-of-medicine/academic-program
  s/graduate-studies/faculty/grad-studies-faculty.cfm?facultyid=423&WT_rank=2