title: Effects of the malononitrilamide FK778 on immune functions in vitro in whole blood from non-human primates and healthy human volunteers.
abstract: BACKGROUND
  FK778, a malononitrilamide analog of leflunomide, is currently being investigated for use in clinical transplantation.
  
  
  METHODS
  Whole blood from cynomolgus monkeys (n=4) and healthy human volunteers (n=4) was incubated with different concentrations of FK778 and stimulated with mitogens in culture medium. Lymphocyte proliferation was assessed by tritium-labeled thymidine incorporation and by flow-cytometric analysis of expression of proliferating cell nuclear antigen on cells in S/G(2)M phase. Flow cytometry was also used to assess expression of T and B lymphocyte activation surface antigens and production of intracellular cytokines by T cells.
  
  
  RESULTS
  Not only lymphocyte proliferation, but also expression of various T cell surface antigens (CD25, CD11a, CD95, CD154) was suppressed by FK778. Fifty percent effective concentration values for the different immune functions were lower in human blood than in blood from cynomolgus monkeys.
  
  
  CONCLUSIONS
  FK778 inhibits multiple immune functions. Their flow cytometric evaluation can be used to assess the effects of the drug in vivo.
authors.affiliation: Department of Cardiothoracic Surgery, Stanford University School of Medicine, 300 Pasteur Drive, Falk CVRB, Stanford, CA 94305-5407, USA.
authors.forename: Tudor
authors.initials: T
authors.lastname: Bîrsan
authors.forename: Camille
authors.initials: C
authors.lastname: Dambrin
authors.forename: Jochen
authors.initials: J
authors.lastname: Klupp
authors.forename: Mario
authors.initials: M
authors.lastname: Stalder
authors.forename: William E
authors.initials: WE
authors.lastname: Fitzsimmons
authors.forename: Randall E
authors.initials: RE
authors.lastname: Morris
mesh_terms: D000480:Alkynes; D000818:Animals; D001681:Biological Assay / Q000379:methods; D001774:Blood Chemical Analysis; D002455:Cell Division / Q000187:drug effects; D005260:Female; D005434:Flow Cytometry; D006801:Humans; D007555:Isoxazoles / Q000494:pharmacology*; D008213:Lymphocyte Activation / Q000187:drug effects*; D008214:Lymphocytes / Q000187:drug effects*; D008297:Male; D009570:Nitriles; D018809:Proliferating Cell Nuclear Antigen / Q000096:biosynthesis
chemical_list: D000480:Alkynes; D007555:Isoxazoles; D009570:Nitriles; D018809:Proliferating Cell Nuclear Antigen; C115284:2-cyano-3-hydroxy-N-(4-(trifluoromethyl)phenyl)-2-hepten-6-ynamide